Document vVk8JRqaewv6kZJnnmg6d5wp9

I 3M Environmental Laboratory FinalReport-AnalyticaSltudy Single-DoseDermal Absorption/ToxicitSytudy ofT-6067,T-6068 and T-6069 in Rabbits In-VivoStudy ReferenceNumber: HWI#6329-152 Study Number: AMDT-01 1095.1 TestSubstance:L-13492 (T-6067,T-6068 and T-6069) Name and Address ofSponsor: 3M SCD Division 367 Grove Street St.Paul,MN 55106 Name and Address ofTestingFacility: 3M EnvironmentalTechnology & Services 935 Bush Avenue -St.Paul,MN 55106 Method Numbers AMDT-M-1-0, AMDT-M-2-0, AMDT-M-4-0, AMDT-M-5-0, AMDT-M-8-0, AMDT-M-14-0, and Revisions: Thermal Extractionof Fluorideby Means of a Modified Dohrmann DX2000 OrganicHalideAnalyzer-Liver FluorideMeasurement by Means of an Orion EA940 Expandable Ion Analyzer Extractionof Fluorocheniicalfsrom Rabbit Liver Analysisof RabbitLiverExtractforFluorochemicalsUsing ElectrosprayMass Spectrometry AnalysisofFluorideUsing theSkalarSegmented Flow Analyzer with Ion SelectiveElectrode Thermal Extractionof Fluorideby Means of a Modified Dohnnann DX2000 Organic HalideAnalyzer-Serum InitiatioDnate: See attachedprotocol Author:James D. Johnson ,@M!oved B . es D. Jo :)n f tmudy DireWr ///Z?/Zf Completion Date wz II)-zgiw @()()001 1.0SUMMARY A t-butyalmmonium salotfperfluorooctanionattheeformoftreatefdabriacndasa liquidformulationwas appliedderrnalltyorabbits.Liversampleswere analyzedat 28 days postdose by combustionfortotalorganicfluorine.The resultsfrom treated animalswere thesame as controlvalues.Alltotalorganicvalueswere below the practicaqluantitatiolnimit.Serum levelswere alsobelow thepracticaqluantitation limitsof theanalysisforsamples collectedatday I and 2 afteradministratioonf the mixtureor thetreatedfabric.From thepharmacokineticstudy(HWI#6329-151), it would be unlikelythatany extentof absorptioncouldhave been detectedinthis study. 2.0 INTRODUCTION This study was designedto provideinformationas to assessthesystemicabsorption and toxicitaynd relativsekinirritancoyf L-13492 (T-6067,T-6068 and T-6069) when appliedto theskinof rabbits. In a pharmacokineticstudy(HWI#6329-15 1),thiscompound showed little accumulationat48 hoursinliver.'The biologicahlalf-lifdeterminedfrom serum levelsinfemalerabbitswas about4 hours afteran intravenousdose. A rabbit injectewdith 40 mg/kg died5 minutesafterinjectionW.ith thisrapidhalf-lifoef disappearance,thereisalmostno chance of being ableto see thematerialaftera dermal dose when the firsstamplingintervails24 hours forserum and 28 days for liver.However, ifthereisa sex differencaend absorptionitispossiblethatthere willbe some appearanceofthematerialinmale rabbits. 3.0 TEST MATERIALS 3.1Test,Control,and Reference Substances and Matrices 3.1.1AnalyticalReference Substance: FC-95, lot161 or 171. They are equivalent. 3.1.2AnalyticalReference Matrix: Bovine liver,bovine serum and rabbit serum 3.1.3Analytical Control Substance: None 3.1.4Analytical Control Matrix: Bovine liver,bovine serum and rabbit serum 3.2 Source of Materials: 3M ICP/PCP Division forFC-95, bovine liverfrom grocerystore,bovine serum from Sigma Chemical Company, rabbitserum from AMDT- 110394.1controlanimals 2 3.3.Purityand StrengtohfReferencSeubstanceR:esponsibiloiftSyponsor. 3.4StabiliotfyReferencSeubstanceT:o be determinebdy Sponsor. 3.5StorageConditionfsorTestMaterialsR:oom temperatufroerFC-95.For biologicalsamples the storageis-2C@+100 C. 3.6 Disposition of Specimens: Biologicaltissuesand fluidswillbe retainedper GLP Regulationforthetimeperiodrequiredforstudieslongerthan28 days. 4.0 EXPER]IMENTAL -Overview The tissuesfrom animalsdosed asdescribed(HWI#6329-152), were availablefor analysisforfluorinecompounds. At the discretioonfthe Study Director,a seriesof analyticatlestscouldbe performed.The screeningforfluorideinlivervia combustion was the most likelyanalysistopresentdefinitivdeataforabsorptionif thepharmacokinetictest(intravenouasdministrationw)as positiveforfluorineinthe liver.Other availabletestswere electrospramyass spectroscopyand gas chromatography/massspectrometryH.owever, ifthedefinitivreesultscouldbe obtainedwith combustionanalysisalone,onlytheliversamples would be analyzed and any othertestswould be forconfirmation.And, ifno organicfluorineincrease was detectablevenjustusingmeter readingsbelow thepracticaqluantitatiolnimit ofthe method, thenfurtheranalysisof liverforspecificfluorinatecdompounds would not be performed. 5.0 EXPERRMENTAL -METHODS 5.1AMDT-M-1-0, Then-nalExtractioonfFluorideby Means ofa Modified Dohnnann DX2000 OrganicHalideAnalyzer-Liver 5.2 AMDT-M-2-0, FluorideMeasurement by Means of an Orion EA940 Expandable Ion Analyzer 5.3 AMDT-M-4-0, Extractionof Fluorochemicalsfrom RabbitLiver 5.4 AMDT-M-5-0, Analysisof RabbitLiverExtractforFluorochemicalsUsing ElectrosprayMass Spectrometry 0()O()O,ll 3 5.5AMDT-M-8-0, AnalysiosfFluoridUesingtheSkalaSregmentedFlowAnalyzer with Ion SelectiveElectrode 5.6 AMDT-M-14-0, Thermal Extractionof Fluorideby Means of a Modified Dohmiann DX2000 OrganicHalideAnalyzer-Serum 6.0 DATA ANALYSIS The dataareattached.There appearstono differenceintotalorganicfluorinein whole liverbetween thecontrolsand the4, 16,40 mg/kg groups. The dose was withrespectto theT-numbered compound. In additiont,hereappearsto be no differencbeetween eitherfabrictreatmentand the controls.The valuesareallbelow thepracticalimitof quantitatioannd thecomparisonsarerelativaend based on meter reading.Very littlteotalorganicfluorinewas observedinserum at I and 2 days postdose ineithersex. The method did not work becausethetime intervaolf samplingwas too late.The materialifpresentwould have been clearedbeforethe samples were collected. Other datawas collectedusingelectrospramyass spectrometry(seeappendices)T.his data,althoughsupportivei,ntheopinionof the Study Directorisnot requiredto reachtheconclusionstatedhere and thereforeisnot discussedindetail. 6.1 Circumstances that May Affectthe Quality of the Data: The possible problem with thisanalysisisthatthereisnota good marker atthe timessamples were collectedtomeasure dermal absorptionifitdid occur.No conclusioncan be drawn from thisdataas totheextentof dermal absorption. 7.0 CONCLUSION In thisstudy,no conclusioncan be drawn from thisdataas tothe extentofdermal absorption. 8.0 MAINTENANCE OF RAW DATA AND RECORDS 8.1 Raw Data and Data: Raw data,approved protocol,approved finalreport, appropriatespecimens,and electronidcatawillbe maintainedintheAMDT archives. 000004 4 9.0 APPENDICES 9.1 Protocoland Amendments 9.1.1Protocoland FinalReportHWI#6329-152 "Single-DoseDermal Absorption/ToxicitSytudy of T-6067,T-6068 and T-6069 inRabbits" (Protocoltype TP3016.AB fordosingof animals,tissuecollectione,tc.) 9.1.2 AnalyticalprotocolAMDT-01 1095.1 9.1.3Amendment toanalyticaplrotocolAMDT-01 1095.1 9.2 Signed Reports from IndividualScientistsN:one 9.3 Quality Assurance Unit Statement: See attached 9.4 Key Personnel Involved inthe Study: See attached 9.5 Materials and Equipment: See methods 9.6 Solutions,Reagents, and St,and-ardsS:ee methods 9.7 Sample Preparation:See methods 9.8 Quality Control Practices:See methods 9.9 Test Methods: See ProtocolAMDT-0 11095.1 9.10 Instrument Settings:See methods 9.11 Data: See attached. 9.11.1Summary and raw data;ug F- inwhole liveras determinedby thermal extractiofnollowedby analysisusing Orion ion analyzer. 9.11.2Sununary and raw data;analysisof liverextractsusingelectrospray mass spectrometry. 0()0()05 5 9.11.3Summary and raw data; ppm F-in serum as determinedby thermal extractionfollowedby analysisusingSkalarsegmented flow analyzerwith ion selectivelectrode. 9.11.4Sununary and raw data;ppm F-in serum as deten-ninedby thermal extractiofnollowedby analysisusingOrion ionanalyzer. 6 9.1.1Protocoland FinalReport HWI#6329-152 "SingleDose Dermal Absorption/ToxicitSytudy of T-6067, T-6068 and T-6069 inRabbits" (ProtocoltypeTP-3016.AB for dosing of animals,tissuecollectione,tc.) 0()0()07 HAZLEWCON W IS C 0 N S IN tvi'Ai) 11.11 i I-, Sponsor: 3M St. Paul, Minnesota i CORNING FINAL REPORT Study Title: Single-Dose Dermal Absorption/Toxicity Study of T-6067, T-6068, and T-6069 in Rabbits Author: Steven M. Glaza Study Completion Date: May 26, 1995 Performing Laboratory: Hazleton Wisconsin, Inc. 3301 Kinsman Boulevard Madison, Wisconsin 53704 LaboratoryProject Identification: HWI 6329-152 Page I of 39 P h o n e 6 0 8 2 -1 71 E @,P R E S S NI A IL D E L E R 'f 3 3 0 1 K ! tl@,f@l-, rj L D Fa til4. -) t (-jI 60 I Page 2 of 39 HWI 6329-152 QUALITY ASSURANCE STATEMENT This report has been reviewed by the Quality Assurance Unit of Hazleton Wisconsin, Inc., in accordance with the Food and Drug Administration (FDA) Good Laboratory Practice Regulations, 21 CFR 58.35 (b) (6) (7). The following inspections were conducted and findings reported to the Study Director and management. Written status reports of inspections and findings are issued to Hazleton management monthly according to standard operating procedures. Inspection Dates From To Phase 11/28/94 12/08/94 02/20/95 05/24/95 05/25/95 11/28/94 12/08/94 02/22/95 05/24/95 05/25/95 Protocol Review Dose Preparation Data/Report Review Report Rereview Report Rereview Date Reported to Study Director Date to Management 11/28/94 12/08/94 02/22/95 05/24/95 05/25/95 12/10/94 01/10/95 03/10/95 06/10/95 06/10/95 c-, Zecilia M. [)@nner Representative, Quality Assurance Unit Date Page 3 of 39 STUDY IDENTIFICATION Single-Dose Dermal Absorption/Toxicity Study of Study of T-6067, T-6068, and T-6069 in Rabbits HWI 6329-152 Test Materials Sponsor Sponsor's Representative Study Director Study Location I T-6067 2 T-6068 3. T-6069 3M Toxicology Service Medical Department 3M Center, Bldg. 220-2E-02 P.O. Box 33220 St. Paul, MN 55133-3220 John L. Butenhoff, PhD 3M Toxicology Service Medical Department 3M Center, Bldg. 220-2E-02 P.O. Box 33220 St. Paul, MN 55133-3220 (612) 733-1962 Steven M. Glaza Hazleton Wisconsin, Inc. P.O. Box 7545 Madison, WI 53707-7545 (608) 241-7292 Hazleton Wisconsin, Inc. Building No. 3 3802 Packers Avenue Madison, Wl 53704 Study Timetable Study Initiation Date Experimental (In-life)Start Date In-life End Date Experimental Termination Date Study Completion Date December 6, 1994 December 8, 1994 January 5, 1995 May 26, 1995 May 26, 1995 0()0()10 Page 4 of 39 HWI 6329-152 KEY PERSONNEL Acute Toxicology Steven M. Glaza Study Director Manager Francis (Bud) W. McDonald Study Coordinator Patricia Padgham In-life Supervisor Rose M. Bridge Report Supervisor Quality Assurance Sherry R. W. Petsel Manager Laboratory Animal Medicine Cindy J. Cary, DVM Diplomate, ACLAM Supervisor Anatomical Pathology Thomas E. Palmer, PhD Anatomical Pathologist Jack Serfort/ Deborah L. Pirkel Supervisors Necropsy Anne Mosher Supervisor Pathology Data Page 5 of 39 CONTENTS Quality Assurance Statement Study Identification Key Personnel Summary Objective Regulatory Compliance Test and Control Materials Test System Procedures Results Discussion Signature Reference Pathology Report Table 1 Individual and Mean Body Weights (g) 2 Individual Clinical Signs 3 Individual Dermal Irritation Scores 4 Individual Pathology Comments Appendix A Protocol TP3016.AB HWI 6329-152 Paqe 2 3 4 6 8 8 8 9 10 13 13 13 14 15 16 18 20 26 28 29 0()0012 Page 6 of 39 HWI 6329-152 SUMMARY This study was done to assess the systemicabsorption/toxicityand relative skin irritancy of T-6067, T-6068, and T-6069 when applied to the skin of rabbits. The study was conducted using three male and three female acclimated rabbits of the Hra:(NZW)SPF strain for each treatment group. Group Dose Level Test Material (mg/kql I (Control) Distilledwater 2 T-6067 3 T-6067 4 T-6067 5 T-6068 5 T-6069 ()a 4 16 40 b b Number of Animals Males Females 3 3 3 3 3 3 3 3 3 3 3 3 a Administered at a do,sevolume of 2.0 mL/kg. b Administered as a 10-cm'x 10-cm piece of test material (fabric). The back of each rabbit was clipped free of hair and a single dose of the respectivematerial at the indicateddose level was administeredto the skin of the rabbits. The treatmentsites remained intact. The area of application was covered with a gauze bandage secured with paper tape around all edges and overwrapped with Saran WrapO and ElastoplastQOtape to provide an occlusive dressing for a 24-hour exposure period. Clinical observations were conducted predose and at approximately 1, 2.5, and 4 hours after test or controlmaterialadministration.Additionalclinical observations and twice a day mortality checks were conducted daily thereafter for 28 days. Body weights were determined on Day -8 or Day -7 for randomization purposes, before test or controlmaterial administration (Day 1), and at in-lifetermination(Day 29). The initialdermal irritation reading was made before test or control material administration(recorded as the Day I reading). Subsequent readings of dermal irritation were made approximately 30 minutes after bandage removal (Day 2) and on Days 4 and 8. Blood samples were collected from a marginal ear vein of the animals before in-life initiation(Day 1), approximately24-hourspostdose (Day 2), on Days 4, 8, 15, and 22. In addition, at the time of necropsy on Day 29, approximately 20 mL of blood was obtained from each animal. All samples were centrifuged and separated into serum and cellular fractions. All animals were euthanized at termination of the in-life phase and necropsied. The whole liver, bile, an approximate 1-cm x 1-cm section of the dermal application site from all animals, and both kidneys from one male and one female in each group were collected at necropsy. The blood samples (serum and cellular fractions), livers, bile, dermal application sites, and kidneys were sent frozen to the Sponsor after termination of the in-life phase. 0()O():L Page 7 of 39 HWI 6329-152 Applicationof T-6067, T-6068, and T-6031 did not result in any test material-relatedchanges in body weight gain or macroscopic findings at necropsy. All animals appearedclinicallynormal throughoutthe study. The control material did not produce any dermal irritation. Test material T-6067 produced very slight irritationin a few of the animalsat all three dose levels (4, 16, and 40 mg/kg). Both fabric test materials,T-6068 and T-6069, produced slight to moderate erythema and slight edema and/or desquamation reactions. Page 8 of 39 HWI 6329-152 OBJECTIVE The objective of this study was to assess the systemic toxicity/absorptionand relative skin irritancyof test materialswhen applied to the skin of rabbits. REGULATORY COMPLIANCE This study was conducted in accordance with the U.S. Food and Drug Administration'sGood Laboratory Practice Regulations for Nonclinical Laboratory Studies, 21 CFR 58. All proceduresused in this study are in compliance with the Animal Welfare Act Regulations. In the opinion of the Sponsor and study director, the study did not unnecessarilyduplicate any previous work. TEST AND CONTROL MATERIALS Identification The test materials were identified a[nddescribed as follows: Identification PhysicalDescrit)tion T-6067 T-6068 T-6069 Clear, colorlessliquid White web cloth White web cloth The control material was distilled water and was described as a clear, colorlessliquid. Purity and Stability The Sponsor assumes responsibilityfor test material purity and stability determinations(includingundertest conditions). The purity and stabilityof the control material were considered to be adequate for the purposes of this study. Storage and Retention The test and control materials were stored at room temperature. A reserve sample of each test and control material was taken and will be retained in a freezer set to maintain a temperatureof -20'C 10'C for 10 years in accordance with Hazleton Wisconsin (HWI) Standard Operating Procedure (SOP). Any unused test material was returned to the Sponsor after completion of all in-life testing according to HWI SOP. Any remaining control material is retained for other testing and will not be discardedafter issuanceof the final report. Page 9 of 39 HWI 6329-152 Safety Precautions The test and control material handling procedures were according to HWI SOPs and policies. TEST SYSTEM Test Animal Adult albino rabbits of the Hra:(NZW)SPFstrain were procured from HRP, Inc., Kalamazoo, MI, on November 16, 1994 and maintained at the Hazleton Wisconsin facility at 3802 Packers Avenue, Madison, Wisconsin. Housing After receipt,the animals were acclimatedfor a period of at least 7 days. During acclimation and throughout the study, the animals were individually housed in screen-bottom stainless steel cages in temperature- and humiditycontrolled quarters. Environme'ntal'controlfsor the animal room were set to maintain a temperature of 19' to 23*C, a relativehumidity of 50% t20%, and a 12-hour light/12-hourdark lightingcycle. In cases where variationsfrom these conditions existed, they were documented and considered to have had no adverse effect on the study outcome. Animal Diet The animals were provided access to water ad libitum and a measured amount of Laboratory Rabbit Diet HF #5326, PMI Feeds, Inc. The feed is routinely analyzed by the manufacturer for nutritional components and environmental contaminants. Samples of the water are periodicallyanalyzed by HWI. There were no known contaminants in the feed or water at levels that would have interferedwith or affectedthe resultsof the study. Selection of Test Animals The animals were identified by animal number and corresponding ear tag and were placed into study groups using a stratified body weight randomization program. The randomization body weights were determined on Day -8. The weight variation of the animals for each group of each sex selected for the study did not exceed t2 standard deviations of the mean weight, and the mean body weights for each group of each sex were not statisticallydifferent at the 5% probabilitylevel. 0()0()1(; Page 10 of 39 Study Design HWI 6329-152 Animals weighing from 2,377 to 2,796 g at initiationof treatmentwere placed into the following study groups: Group Dose Level Number of Animals Test Material -(mg/kq)- Males Females I (Control) Distilledwater oa 3 3 2 T-6067 4 3 3 3 T-6067 16 3 3 4 T-6067 40 3 3 5 T-6068 b 3 3 6 T-6069 b 3 3 a Administered at a dose volume of 2.0 mL/kg. b Administered as a 10-cm x 10-cm section of test material(fabric). Justificationfor Species Selection Historically, the New ZealandWhite albino rabbit has been the animal of choice because of the large amount of background informationon this species. PROCEDURES Preparation of Exposure Area On the day before test material application,the back and, if necessary (to obtain unblemishedskin), the flanks of each rabbit was clipped free of hair. The clipped area made up approximately 20% of the total body surface area. The test sites (intactskin)were inspectedfor interferinglesions, irritation,or defects that would precludethe use of any of the animals. The animals were clipped on Days 8 and 29 to aid in visualizingthe application sites. Dose Administration All animals receiveda single administrationof the respectivetest or control material. The day of treatment was designated as Day 1. Group 1. An individual dose (2.0 mL/kg) was calculated and measured based on each animal's body weight on the day of treatment. The control material (distilledwater) was appliedevenly to the test site at a rate of approximately 0.05 mL/cm 0()O()IL7 Page 11 of 39 HWI 6329-152 Groups 2, 3, and 4. The test material (T-6067) was administered undiluted to the test sites of the Groups 2, 3, and 4 animals (4, 16, or 40 mg/kg, respectively) using the average bulk density of 1.04 g/mL to determine the dose volume for each dose level. An individual dose of the test material was calculated for each animal based on its body weight on the day of treatment. The area of exposure for the 4@ 16, and 40 mg/kg dose levels was 9, 9, and 16 cm2, respectively. The approximate rate of application ranged from 0.001 to 0.006 mL/cm2 . Groups 5 and 6. The test material (T-6068 or T-6069) was applied to the respective test site as a 10-cm x 10-cm section of material that was moistened with distilled water. Each area of application was covered with a 10-cm x 10-cm gauze bandage secured with paper tape around all edges and overwrapped with Saran Wrapo and ElastoplastO tape to provide an occlusive dressing. Collars were used to restrain the animals during the 24-hour exposure period. Approximately 24 hours after test or control material application, the restraining collars and bandages"wey-eremoved and the residual test material was removed with tap water and disposable paper towels. Reason for Route of Administration The dermal route is a potential route of exposure in humans. Observations of Animals Clinical observations were conducted predose and at approximately 1, 2.5, and 4 hours after test or control material administration. Additional clinical observations and twice a day mortality checks (morning and afternoon) were conducted daily thereafter for 28 days. Body weights were determined for randomization purposes on Day -8 or Day -7, before test material administration(Day 1), and at in-life termination (Day 29). The initial dermal irritation reading was made before test or control material administration according to the Draizel technique (recorded as the Day 1 reading). Subsequent readings of dermal irritation were made approximately 30 minutes after bandage removal (Day 2) and on Days 4 and 8. Page 12 of 39 Sample Collections HWI 6329-152 Blood samples (approximately 4 mL) were collected from a marginal ear vein of all animals before experimental initiation (Day 1). Subsequent collection of blood was conducted 24-hours postdose (Day 2), and on Days 4, 8, 15, and 22. In addition, at the time of necropsy on Day 29, approximately 20 mL of blood was obtained from the posterior vena cava of each animal. All samples were centrifuged and separated into serum and cellular fractions. These samples were then stored in a freezer set to maintain a temperature of -200C 100C until shipped to the Sponsor. Pathology At termination of the experimental phase (Day 29), animals were anesthetized with sodium pentobarbital, bled via the posterior vena cava, exsanguinated, and necropsied in random order. The sites of test and control material application were washed with lukewarm tap water before the necropsy procedure. All animals were subjected to an abbreviated gross necropsy examination and any abnormalities were recorded., The whole liver, bile, an approximate 1-cm x 1-cm section of the dermal application site from all animals, and both kidneys from the first male and female in each group were collected and immediately placed on dry ice, then placed in a freezer set to maintain a temperature of -200C 10'C. After necropsy, the animals were discarded. Shi-pmentof Blood, Bile, and Tissues After experimental termination, the blood samples (serum and cellular fractions), livers, bile, dermal application sites, and kidneys were sent frozen (on dry ice) to the Sponsor (James D. Johnson, 3M E.E. & P.C., Bldg. 2-3E-09, 935 Bush Avenue, St. Paul, MN, 55106). The Sponsor is responsible for the retention and disposition of the samples. HWI does not accept any responsibility for the analysis of the tissue samples collected in this study nor are these results presented in this report. Statistical Analyses No statistical analyses were required by the protocol. Location of Raw Data, Records, and Final ReDort The raw data, records, and an original signed copy of the final report will be retained in the archives of HWI in accordance with HWI SOP. Page 13 of 39 Body Weights RESULTS HWI 6329-152 Individual and mean body weights are in Table 1. All animals exhibited body weight gains from Day I to Day 29. Clinical Observations Individualclinical signs are in Table 2. All animals appearednormal throughout the study. Dermal Irritation Individualdermal irritationscoresare in Table 3. The control material produced no dermal irritation. Test material T-6067 produced slight erythema reactions in one animal treated at 4 mg/kg, in two animals treated at 16 mg/kg, and in two animals treated at 40 mg/kg. Both fabric test materials, T-6068 and T-6069, produced slight to moderate erythema and slight edema and desquamation reactions. Pathology Individual animal pathology comments are presented in Table 4. There were no lesions observed in any of the animals. Page 15 contains a pathology report by the study pathologist. DISCUSSION The acute systemic absorption/toxicityand relative skin irritancy of T-6067, T-6068, and T-6069 were evaluated in male and female albino rabbits when administered as a single dermal application. Applicationof the these materials did not result in any effects on body weight gain or macroscopic findings at necropsy. All animals appearednormal throughout the study. No dermal irritationwas observedas a result of the applicationof the control material,very slight irritationwas seen in a few of the animals treatedwith T-6067, and slight to moderate irritationwas seen in the animals treated with T-6068 or T-6069. SIGNATURE Steven M. Glaza Study Director Acute Toxicology !@@ Date Page 14 of 39 HWI 6329-152 REFERENCE 1. Draize, J. H., "Acute Dermal Toxicity (Single Exposure)," In: Appraisal of the Safety of Chemicals in Foods, Drugs and Cosmetics - Dermal Toxicity, Association of Food and Drug Officials of the U.S., pp. 54-56 (1959). Page 15 of 39 PATHOLOGY REPORT HWI 6329-152 There were six rabbits (three males and three females) each from six dose levels euthanized and necropsiedat the terminationof the study. The test material, dose level, day of death, and gross observations recorded for each animal are in the IndividualPathologyComments that follow this report. At necropsy,there were no visible lesions in any of the animals. The liver, bile, an approximate 1-cm x 1-cm section of the dermal application site from all animals, and both kidneys from one male and one female in each group were collected,frozen, and sent to the Sponsor. After necropsy, the animals were discarded. Th-omYs-E. -P-alm-er,--PhV- Date Pathologist (6329-152.lhm) 020895 Animal Number F52725 F52711 F52719 Mean Page 16 of 39 HWI 6329-152 Table I Individualand Mean Body Weights (g) Male Randomization Day -8 Day 1 29 Animal Number Female Random- ization Day Day -8 1 29 Group I (Control)- DistilledWater (0 mg/kq) 2,699 2,592 2,456 2,582 2,796 2,726 2,494 2,672 3,135 3,085 2,671 2,964 F52721 F52806 F52727 2,463 2,524 2,639 2,542 2,651 2,627 2,713 2,664 2,963 3,086 3,103 3,051 F52718 F52723 F52713 Mean 2,380 2,482 2,602 2,488 Group 2 - T-6067 (4 mg/kq) 2,450 2,778 2,527 -2,795 .3,181 2,980 F52715 F52809 F52738 2,363 2,498 2,489 2,585 2,985 2,450 2,377 2,511 2,619 2,502 2,752 2,909 2,946 2,869 F52737 F52801 F52724 Mean 2,631 2,611 2,634 2,625 Group 3 - T-6067 (16-mg/kq) 2,634 2,699 2,616 2,878 3,113 2,933 F52733 F52788 F52720 2,543 2,575* 2,560 2,650 2,975 2,559 2,629 2,429 2,627 2,562 3,017 2,994 3,028 3,013 F52802 F52730 F52731 Mean 2,543 2,408 2,339 2,430 Group 4 - T-6067 (40 mg/kq) 2,785 2,520 2,489 3,088 2,766 2,715 F52714 F52734 F52774 2,523 2,557 2,364* 2,598 2,856 2,481 2,578 2,660 2,556 2,598 2,893 2,839 2,930 2,887 Day -7 body weight. OOOOZ.3 Animal Number F52729 F52803 F52712 Mean Page 17 of 39 HWI 6329-152 Table I (Continued) Individual and Mean Body Weights (g) Male Randomization Day -8 Day 1 29 Animal Number Female Random- ization Day Day -8 -1 29 Group 5 - T-6068 (10 cm x 10 cm Section) 2,431 2,442 2,631 2,545 2,596 2,645 2,893 2,920 2,886 F52716 F52787 F52728 2,652 2,494* 2,495 2,778 2,544 2,619 3,211 3,017 3,009 2,501 2,595 2,900 2,547 2,647 3,079 F52736 F52717 F52735 Mean Group 6 - T-6069 (10 cm x 10 cm Section) 2,681 2,405 2,570 2,745 2,538 2,606 -2,996 3,025 2,875 F52739 F52732 F52726 2,442 2,477 2,591 2,552 2,630 2,965 2,503 2,480 2,525 2,768 2,591 2,957 2,930 3,110 2,999 Day -7 body weight. Sex Male Female Male Female Male Female Page 18 of 39 HWI 6329-152 Table 2 IndividualClinical Signs Animal Number Observation 1-4 Hours (Day I-L Day 2 through 29 Group I (Control)- DistilledWater (0 mg/kq) F52725 Appeared nc)rmal F52711 Appeared normal i F52719 Appeared normal F52721 Appeared normal F52806 Appeared normal F52727 Appeared normal Group 2 -'T-6067 (4 mg/kq) F52718 Appeared normal F52723 Appeared normal F52713 Appeared normal F52715 Appeared normal F52809 Appeared normal F52738 Appeared normal Group 3 - T-6067 (16 mg/kq) F52737 Appeared normal F52801 Appeared normal F52724 Appeared normal F52733 Appeared normal F52788 Appeared normal F52720 Appeared normal Condition existed. 0()00;:5 Sex Male Female Male Female Male Female Page 19 of 39 HWI 6329-152 Table 2 (Continued) Individual Clinical Signs Animal Number Observation 1-4 Hours (Day 1) Day 2 through 29 Groui)4 - T-6067 (40 mg/kq) F52802 Appeared normal F52730 Appeared normal F52731 Appeared normal F52714 Appeared normal F52734 Appeared normal F52774 Appeared normal Group 5 - T-6068^(10 cm x 10 cm Section) F52729 Appeared normal F52803 Appeared normal F52712 Appeared normal F52716 Appeared normal F52787 Appeared normal F52728 Appeared normal Group 6 T-6069 (10 cm x 10 cm Section) F52736 Appeared normal F52717 Appeared normal F52735 Appeared normal F52739 Appeared normal F52732 Appeared normal F52726 Appeared normal Condition existed. Page 20 of 39 Table 3 IndividualDermal IrritationScores HWI 6329-152 Group I (Control)- DistilledWater (0 mg/kg) Dermal Reaction Erythema Edema Atonia Desquamation Coriaceousness Fissuring Males Study Day 1 2 4 8 Animal No. F52725 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Females Study Day 1 2 4 -8 Animal No. F52721 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Erythema Edema Atonia Desquamation Coriaceousness Fissuring -Animal No. F52711 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Animal No. F52806 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Erythema Edema Atonia Desquamation Coriaceousness Fissuring Animal No. F52719 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Animal No. F52727 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0000%7 Page 21 of 39 HWI 6329-152 Table 3 (Continued) IndividualDermal IrritationScores Group 2 - T-6061 (4 mg/kg) Dermal Reaction Males Study Day 1 2 4 8 Females Study Day 1 2 4 8 Animal No. F52718 Animal No. F52715 Erythema Edema Atonia Desquamation Coriaceousness Fissuring 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Erythema Edema Atonia Desquamation Coriaceousness Fissuring Animal No. F52723 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Animal No. F52809 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Erythema Edema Atonia Desquamation Coriaceousness Fissuring Animal No. F52713 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Animal No. F52738 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Page 22 of 39 HWI 6329-152 Table 3 (Continued) IndividualDermal IrritationScores Dermal Reaction Erythema Edema Atonia Desquamation Coriaceousness Fissuring Group 3 - T-6067 (16 mg/kg) Males Study Day 2 8 Females Study Day 1 2 4 8 Animal No. F52737 Animal No. F52733 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Erythema Edema Atonia Desquamation Coriaceousness Fissuring Animal No. F52801 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Animal No. F52788 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Erythema Edema Atonia Desquamation Coriaceousness Fissuring Animal No. F52724 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Animal No. F52720 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0()OOZ9 Page 23 of 39 HWI 6329-152 Table 3 (Continued) IndividualDermal IrritationScores Group 4 - T-6067 (40 mg/kg) Dermal Reaction Males Study Day 1 2 4 -8 Animal No. F52802 Females Study Day 1 2 4 -8 Animal No. F52714 Erythema Edema Atonia Desquamation Coriaceousness Fissuring 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Erythema Edema Atonia Desquamation Coriaceousness Fissuring Animal No. F52730 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Animal No. F52734 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Erythema Edema Atonia Desquamation Coriaceousness Fissuring Animal No. F52731 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Animal No. F52774 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0000,30 Page 24 of 39 HWI 6329-152 Table 3 (Continued) IndividualDermal IrritationScores Group 5 - T-6068 (10 cm x 10 cm Section) Dermal Reaction Males Study Day 1 2 4 8 Females Study Day 1 2 4 8 Animal No. F52729 Animal No. F52716 Erythema Edema Atonia Desquamation Coriaceousness Fissuring 0 1 1 1 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Erythema Edema Atonia Desquamation Coriaceousness Fissuring Animal No. F52803 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Animal No. F52787 0 1 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Erythema Edema Atonia Desquamation Coriaceousness Fissuring Animal No. F52712 0 1 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Animal No. F52728 0 1 2 1 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0()0031 Page 25 of 39 HWI 6329-152 Table 3 (Continued) IndividualDermal IrritationScores Group 6 T-6069 (10 cm x 10 cm Section) Dermal-Reaction Males Study Day 4 8 Females Study Day 1 2 4 8 Animal No. F52736 Animal No. F52739 Erythema Edema Atonia Desquamation Coriaceousness Fissuring 0 1 2 1 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 1 2 1 0 1 1 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 Erythema Edema Atonia Desquamation Coriaceousness Fissuring Animal No. F52717 0 1 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Animal No. F52732-. 0 1 1 0 0 1 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Erythema Edema Atonia Desquamation Coriaceousness Fissuring Animal No. F52735 0 1 2 1 0 1 1 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 Animal No. F52726 0 1 2 1 0 0 1 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 ()()0032 Animal Number F52725 F52711 F52719 F52721 F52806 F52727 F52718 F52723 F52713 F52715 F52809 F52738 F52737 F52801 F52724 F52733 F52788 F52720 Page 26 of 39 HWI 6329-152 Table 4 Individual Pathology Comments Test Day Sex Died Sacrificed Necropsy Observation Group I (Control) - Distilled Water (0 mg/kq) m 29 No visible lesions. m - 29 No visible lesions. m - 29 No visible lesions. F - 29 No visible lesions. F - F - 29 No visible lesions. 29 No visible lesions. Grout)"2 -,T-6067 (4 mg/kq) m - m - 29 No visible lesions. 29 No visible lesions. m - F - F - 29 No visible lesions. 29 No visible lesions. 29 No visible lesions. F - 29 No visible lesions. Group 3 - T-6067 (16 mg/kq) m - m - m - 29 No visible lesions. 29 No visible lesions. 29 No visible lesions. F - 29 No visible lesions. F - 29 No visible lesions. F - 29 No visible lesions. - Not applicable. Page 27 of 39 Table 4 (Continued) Individual Pathology Comments HWI 6329-152 Animal Test Day Number Sex Died Sacrificed Necropsy Observation F52802 m F52730 m F52731 m F52714 F F52734 F F52774 F Group 4 - T-6067 (40 mqlkq) - 29 No visiblelesions. - 29 No visiblelesions. - 29 No visible lesions. - 29 No visible lesions. - 29 No visible lesions. - 29 No visible lesions. Group 5 - T-6068,(10 cm x 10 cm Section) F52729 m - 29 No visible lesions. F52803 m - 29 No visible lesions. F52712 m - 29 No visible lesions. F52716 F - 29 No visiblelesions. F52787 F - 29 No visiblelesions. F52728 F - 29 No visiblelesions. F52736 F52717 F52735 F52739 F52732 F52726 Group 6 T-6069 (10 cm x 10 cm Section) m - 29 No visible lesions. m - 29 No visible lesions. m - 29 No visiblelesions. F - 29 No visible lesions. F - 29 No visiblelesions. F - 29 No visible lesions. - Not applicable. 000034 Page 28 of 39 APPENDIX A Protocol TP3016.AB I HWI 6329-152 0()0035 HAZLE'ZC:N W IS C 0 N S IN Nl A 1)1'. r, VI I I fI 1',,l1. page 29 of 39 CORNING Sponsor: 3M St. Paul, Minnesota PROTOCOL TP3016.AB Study Title: Single-DoseDermal Absorption/ToxicityStudy of T-6067, T-6068, and T-6069 in Rabbits Date: December 6, 1994 Performing Laboratory: Hazleton Wisconsin, Inc. 3301 Kinsman Boulevard Madison, Wisconsin 53704 LaboratoryProjectIdentification: HWI 6329-152 p ti U!, E x F'@@ ii 6 2,1 .1 NI -%'L L)E L iv I F ,Uri 2,. 0()003(; Page 30 of 39 STUDY IDENTIFICATION TP3016.AB Page 2 Single-DoseDermal Absorption/ToxicityStudy of T-6067, T-6068, and T-6069 in Rabbits HWI No. Test Materials Sponsor Sponsor's Representative Study Director Study Location Proposed Study Timetable Experimental Start Date Experimental Termination Date Draft Report Date 6329-152 1. T-6067 2. T-6068 3. T-6069 3M Toxicology Services 220-2E-02 3M Center St. Paul, MN 55144 John L. Butenhoff, PhD 3M Toxicology Services 220-2E-02 3M Center St. Paul, MN 55144 (612) 733-1962 Steven M. Glaza Hazleton Wisconsin, Inc. P.O. Box 7545 Madison, Wl 53707-7545 (608) 241-7292 Hazleton Wisconsin, Inc. BuildingNo. 3 3802 Packers Avenue Madison, WI 53704 December 8, 1994 January 5, 1995 February 16, 1995 000037 Page 31 of 39 TP3016.AB Page 3 1. Study Single-DoseDermal Absorption/ToxicityStudy in Rabbits 2. Purpose To assess the systemicabsorptionand toxicity and relative skin irritancyof test materialswhen appliedto the skin of rabbits 3. Regulatory Comi)liance This study will be conducted in accordancewith the followingGood LaboratoryPractice Regulations/Standards/Guidelines: f ) Conduct as a Nonregulated Study (X] 21 CFR 58 (FDA) 40 CFR 160 (EPA-FIFRA) 40 CFR 792 (EPA-TSCA) C(81)30(Final)(OECD) 59 Nohsan No. 3850 (Japanese MAFF) NotificationNo. 313 (JapaneseMOHW) All proceduresin this protocolare in compliancewith the Animal Welfare Act Regulations. in tfieopinionof the Sponsor and study director, the study does not unnecessarilyduplicate any previous work. 4. Ouality Assurance The protocol,study conduct,and the final reportwill be auditedby the Quality AssuranceUnit in accordancewith Hazleton Wisconsin (HWI) StandardOperating Procedures (SOPS) and policies. 5. Test Materials A. Identification T. T-6067 2. T-6068 3. T-6069 B. Physical Description 1. (To be documentedin the raw data) 2. (To be documentedin the raw data) 3. (To be documentedin the raw data) C. Purity and Stability The Sponsorassumesresponsibilityfor purity and stability determinations(includingunder test conditions). Page 32 of 39 TP3016.AB Page 4 D. Storage Room temperature E. Reserve Samples Reservesample(s)of each batch/lotof test and control materialswill be taken for this study. The test and controlmaterialreservesampleswill be stored at HWI in a freezer set to maintain a temperatureof -20*C tlO*C for 10 years per HWI SOP. The Sponsor will be contacted after 10 years for dispositionin accordancewith the appropriate regulatoryGood LaboratoryPractices. F. Retention Any unused test materialswill be returnedto the Sponsorafter completionof the in-lifephase of the study. G. Safety Precautions As required by HWI SOPs and policies 6. Control Material A. Identification bistilledwater B. Physical Description Clear,colorlessliquid C. Purity and Stabilit @he purity and stabilityof this manufacturedmaterialis consideredto be adequatefor the purposes of this study. D. Storage Conditions Room temperature E. Reserve Samples See Section 5. E. Reserve Samples F. Retention @ny remaining controlmaterial may be used for other testing and will not be discardedafter issuanceof the final report. G. Safety Precautions As required by HWI SOPs and policies OoOO39 Page 33 of 39 TP3016.AB Page 5 7. ExperimentalDesign A. Animals (1) Species Rabbit (2) Strain/Source Hra:(NZW)SPF/HRP, Inc. (3) Age at Initiation Adult (4) Weight at Initiation 2.0 to 3.0 kg (5) Number and-Sex 18 males and 18 females (6) Identification Individualnumbered ear tag (7) Husbandry (a) Housing Individually,in screen-bottomstainlesssteel cages (heavy gauge) (b) Food A measured amount of Laboratory Rabbit Diet HF #5326 (PMI Feeds, Inc.). The food is routinelyanalyzedby the manufacturerfor nutritionalcomponents and environmental contaminants. (c) Water Ad libitum from an automatic system. Samples of the water are analyzedby HWI for total dissolved solids, hardness, and specified microbiologicalcontent and for selected elements, heavy metals, organophosphates,and chlorinated hydrocarbons. (d) Contaminants There are no known contaminants in the food or water that would interferewith this study. (e) Environment Environmentalcontrolsfor the animal room will be set to maintain a temperatureof 19*C to 23'C, a relative humidity of 50% +20%, and a 12-hour light/12-hourdark cycle. 0()00.10 Page 34 of 39 TP3016.AB Page 6 (f) Acclimation At least 7 days (8) Selectionof Test Animals Based on health and body weight according to HWI SOPS. An adequate number of extra animals will be purchased so that no animal in obviously poor health is placed on test. The animalswill be placed into study groups using a stratifiedbody weight randomizationprogram within nine days of study initiation. (9) Justificationfor S2eciesSelection Historically,the New Zealand White albino rabbit has been the animal of choice because of the large amount of background informationon this species. B. Dose Administration (1) Test Groups Group Test Material I (Control)Distilledwater 2 T-6067 3 T-6067 4 T-6067 5 T-6068 6 T-6069 Dose Level (mg/kg) 0* 4 16 40 Number of Animals Males Females 3 3 3 3 3 3 3 3 3 3 3 3 To be administeredat a dose volume of 2.0 mL/kg To be administeredas a 10.0-cmx 10.0-cmpiece of test material(fabric) (2) Preparation of Exposure Area On the day before test material application,the back and, if necessary(to obtainunblemishedskin), the flanksof each rabbitwill be clippedfree of hair with an electric clipper. The shaved area will constituteapproximately 20% of the total body surfacearea. The treatmentsites (intactskin)will be inspectedfor interferinglesions, irritation,or defectsthat would precludethe use of any of the animals. The animalswill be clipped as needed throughout the study. Page 35 of 39 TP3016.AB Page 7 (3) Dose Administration All animalswill receive a,singleadministrationof the respectivetest or controlmaterial. The day of treatment will be designatedas Day 1. The respectivedoses for the animals in Groups 1, 2, 3, and-4 will be based on the animal'sbody weight just before administrationand spread onto the area of exposurein a thin and uniform layer. The Group 1, 2, 3, and 4 materialswill be applied undiluted. The Group 5 and 6 materialswill be appliedas a 10.0-cmx 10.0-cmpiece of the respectivetest material moistenedwith distilledwater. The area of application (Groups 1-6) will be covered with a 10-cm x 10-cm gauze bandage secured with paper tape around all edges and overwrappedwith Saran Wrap* and Elastoplast*tape to providean occlusivedressing. The rabbitswill be collaredduring the 24-hourapplicationperiod. (4) Reason for Route of Administration The dermal route is a potentialroute of exposure in humans. (5) Removal of Test Material Approximately24 hours after test or controlmaterial applicationthe bandagesand collarswill be removed and the residualtest materialwill be removedusing water or an appropriatesolvent, if necessary. C. Observationof Animals (1) Clinical Observations For clinicalsignsbefore test or controlmaterial administrationand for clinicalsigns and mortalityat approximately1, 2.5, and 4 hours after test material administration(Day 1) and dailythereafterfor clinical signs,and twicedaily (a.m.and p.m.)for mortalityfor at least 28 days. Observationsmay be extended when directedby the study director. (2) Reading of Dermal Irritation Before test or controlmaterialadministrationthe initial dermal irritationreadingwill be made and recorded as the Day I reading (Attachment1). Additionaldermal irritationreadingswill be made approximately30 minutes after bandage removal (Day 2) and on Study Days 4 and 8. Individualdermal irritationrecordswill be maintained for each animal. (3) Body Weights Just beforetes" or controlmaterialapplication(Day 1), on Day 29, and at unscheduleddeath (when survival exceeds I day) Page 36 of 39 TP3016.AB Page 8 (4) SampleCollections (a) Frequency Before initiation(Day 1), approximately24 hours post-dose(Day2), Days 4,.8, 15, 22, and at experimentaltermination(Day 29) (b) Number of Animals All (c)Method of Collection Blood samples (approximately4 mL) will be collected from the marginalear vein of either ear on Days 1, 2, 4, 8, 15, and 22. Approximately20 mL of blood (actualvolumeto be documentedin the raw data) will be obtained from the posterior vena cava of each animalsacrificedin a moribundconditionor sacrificedat the time of necropsy(Day 29). The samples will be stored at room temperature and then centrifuged,and the separate serum and cellular fractionsstoredin a freezer set to maintain -20*C tlO*C. The separated serum and cellular fractionswill be sent frozen to the Sponsor after experimentaltermination. Samples will be shipped to: James D. Johnson 3M E.E. & P.C. Bldg. 2-3E-09 935 Bush Avenue St. Paul, MN 55106 James D. Johnson or alternatewill be notified by telephoneat (612)778-5294prior to the shipmentof the samples. D. Pathology (1) Unscheduled Sacrifices and Deaths Any animaldying during the study or sacrificedin a moribund conditionwill be subjectedto an abbreviated gross necropsyexaminationand all abnormalitieswill be recorded. Animalsin a moribundconditionwill be anesthetizedwith sodiumpentobarbital(via injectionin the marginalear vein), bled via the vena cava, and exsanguinated.After necropsy,the animalswill be discarded. 000043 Page 37 of 39 TP3016.AB Page 9 (2) Scheduled Sacrifice At terminationof the experimentalphase (Day 29), survivinganimalswill be anesthetizedwith sodium pentobarbital(via injectionin the marginalear vein), bled via the vena cava, exsanguinated,and subjected to an abbreviatedgross necropsyexamination. The animals will be necropsiedin randomorder and all abnormalitieswill be recorded. (a) Sample Collection The sitesof test and controlmaterialapplication will be washed with lukewarmtap water prior to the necropsyprocedure. The whole liver,bile, an approximate 1-cm x 1-cm section of the dermal applicationsite from all animals,and both kidneys from the first male and female necropsied in each group will be collectedand immediatelyplaced in a freezer set to maintaina temperatureof -20*C tlO*C. After necropsy,the animalswill be discarded. The tissues..(Iivebri,le,dermalapplicationsite, kidneys)will be sent frozenon dry ice to the Sponsor after experimentaltermination. The samples will be shippedto the personlisted in Section 7.C.(4).(c).The Sponsoris responsiblefor the retentionand dispositionof the samples. E. StatisticalAnalyses No statisticalanalysesare required. 8. Report A final report includingthose items listedbelow will be submitted. Descriptionof the test and controlmaterials Descriptionof the test system Procedures Dates of experimentalinitiationand termination Tabulationof mortalitydata by sex and dose level Descriptionof any toxic effects/dermalirritation Tabulation of mean body weights by sex and dose level Gross pathology findings/grosspathology report oo0044 Page 38 of 39 TP3016.AB Page 10 9. Location of Raw Data, Records, and Final ReDort Originaldata, or copies thereof,will be availableat HWI to facilitateauditingthe studyduring its progressand before acceptanceof the finalreport. When the final report is completed, all originalpaper data, includingthose item listed below will be retained in the archivesof HWI according to HWI SOP. Protocol and protocol amendments Dose preparation records In-liferecords Body weights Dose administration Observations Anatomical pathology records Sample collectionrecords Shipping records Study correspondence Final report(originalsignedcopy) The followingsupportingrecordswill be retainedat HWI but will not be archivedwith the siudydata. Animal receipt/acclimationrecords Water analysis records Animal room temperature and humidity records Refrigeratorand freezer temperaturerecords Instrumentcalibrationand maintenance records 000045 Page 39 of 39 PROTOCOL APPROVAL TP3016.AB Page 11 John L. Butenhoff, PhD Sponsor's Representative 3M Steven M. Glaza Study Director Acute Toxicology Hazleton Wisconsin, Inc. L;@x@@s-entative Quality Assurance Unit Hazleton Wisconsin, Inc. (6329-152.protdskl) -2-Date '5a-te Date oo0046 9.1.2 AnalyticalprotocolAMDT-01 1095.1 000047 3M EnvironmentalLaboratory Protoco-lAnalyticaSltudy Single-DosDermalAbsorption/ToxicSittuydyofT-6067,T-6068and T-6069 inRabbits In-VivoStudyReferencNeumber:HWI#6329-152 StudyNumber: AMDT-01 1095.1 TestSubstanceL:-13492(T-6067T,-6068andT-6069) Name and AddressofSponsor: 3M SCD Division 367 Grove Street St.Paul,MN 55106 Name and Address of TestingFacility: 3M EnvironmentalTechnology and Services 935 Bush Avenue St.Paul,N4N 55106 Proposed InitiationDate: July25, 1995 Proposed Completion Date: August 25, 1995 Method Numbers AMDT-M-1-0, AMDT-M-2-0, AMDT-M4-0, AMDT-M-5-0, AMDT-M-8-0, AMDT-M-14-0, and Revisions: Thermal Extractionof Fluorideby Means of a Modified Dohrmann DX2000 OrganicHalideAnalyzer-Liver FluorideMeasurement by Means ofan Orion EA940 Expandable Ion Analyzer ExtractionofFluorochen-iicaflrsom RabbitLiver Analysisof RabbitLiverExtractforFluorochemicalsUsing ElectrosprayMass Spectrometry AnalysisofFluorideUsing the SkalarSegmented Flow Analyzer with Ion SelectiveElectrode Thermal Extractionof Fluorideby Means of a Modified Dohrinann DX2000 OrganicHalideAnalyzer-Serum Author:James D. Johnson rovejd By- ove, m ers D. @@/son //Study Dit@@6tor Date John Butenhoff,PhD Date Sponsor Representative ()()00119 1.0 PURPOSE ThisstudyisdesignetdoprovidienformatiaosntowhetherL-13492(T-6067, T-6068 and T-6069) isdennallyabsorbed.The analyticaalspectof thisstudyisto determinetotalorganicfluorineinthetissueand serum of rabbitsatvarioustimes postdose dermal applicatioonfL-13492. 2.0 TEST MATERIALS 2.1Test,Control,and Reference Substances and Matrices 2.1.1AnalyticalReference Substance: FC-95, lot161 or 171. They are equivalent. 2.1.2AnalyticalReference Matrix: Bovine liverand bovine serum 2.1.3AnalyticalControl Substance: None 2.1.4AnalyticalControl Matrix: Bovine liverand bovine serum 2.2 Source of Materials:3M ICP/PCP Division(2.1.1),Grocery store(2.1.2,2.1.4 liver)S,igma Chemical Company (2.1.22,.1.4serum) 2.3Number of Test and Control Samples: Tissuesand fluidsfrom 30 testanimals and 6 controlanimals.Tissuesand fluidsincludeliverk,idney,serum,cellular fractiond,ermal applicatiosniteand bile.Analysisofthesetissueswillbe atthe discretioonf the Study Director. 2.4Identificatioonf Test and Control Samples: The samplesare identifieudsing the HWI animalidentificatinounmber which consistsof a lettearnd fivedigit number, plusthetissueidentitaynd day identit(yserum). 2.5 Purity and Strength ofReference Substance: To be determinedby Sponsor. 2.6 Stabilityof Reference Substance:To be determinedby Sponsor. 2.7 Storage Conditions forTest Materials:Room temperature(2.1.1), -20 10-C (2.1.22,.1.4).Testand Controlsampleswillbe receivedaccordingto AMDT-S-10-0. 2.8Dispositionof Specimens:Biologicatlissueasnd fluidswillbe retainedper GLP Regulationforthetimeperiodrequiredforstudieslongerthan28 days. 2.9 SafetyPrecautions:Refer toappropriateMSDS. Wear appropriatelaboratory attireU.se cautionwhen handlingknivesforcuttingthesamples. 2 0()004!) 3.0 EXPERIMENTAL -Overview The tissuesftom animalsdosed as described(HWI#6329-152), areavailablefor analysisforfluorinecompounds. At the discretioonf theStudy Director,a seriesof analyticatlestscan be performed. The screeningfor fluoridein livervia combustion (See Methods--next Section)isthe appropriateanalysisto presentdefinitivdeatafor fluorineinthe liver. Not allof thetissuesand fluidsamples willbe analyzed. When sufficiendtata has been collectedto meet the objectivesof the study in the opinionof the Study Director,analysiswillcease. 4.0 EXPERIMENTAL - Methods 4.1Liver and Serum screeningmethods: (attached) 4.1.1AMDT-M-1-0, Thermal Extractionof Fluorideby Means of a Modified Dohrinann DX2000 Organic Halide Analyzer-Liver 4.1.2AMDT-M-2-0, FluorideMeasurement by Means of an Orion EA940 Expandable Ion Analyzer 4.1.3AMDT-M-4-0, Extractionof Fluorochemicalsfrom Rabbit Liver 4.1.4AMDT-M-5-0,, Analysisof Rabbit LiverExtractforFluorochemicals using ElectrosprayMass Spectrometry 4.1.5AMDT-M-8-0, Analysisof FluorideUsing the SkalarSegmented Flow Analyzer with Ion SelectiveElectrode 4.1.6AMDT-M-14-0, Thermal Extractionof Fluorideby Means of a Modified Dohrmann DX2000 Organic HalideAnalyzer-Serum 5.0DATA ANALYSIS 5.1Data Reporting: Data willbe reportedasa concentratio(nweight/weighto)f fluorideper tissueor fluid,or as FC-143 (electrospramyass spectrometry)per unit of tissueor fluid.Statisticussed,atthediscretioonf theStudy Director,may includeregressionanalysisof serum concentrationswith time and averages and standarddeviationsof concentrationfsor differendtose groups. Ifnecessary(inthe opinionof the Study Director),simple statistictaelstssuch as Student'st testmay be appliedto determine statisticdailfference. 3 0()0050 6.0 MAINTENANCE!12F RAW DATA AND RECORDS 6.1 Raw Data and Records: Raw data,approvedprotocol,appropriatespecimens, approved finalreport,and electronidcatawillbe maintainedinthe AMDT archives. 7.0 REFERENCES 7.1AMDT-S-10-0, Sample TrackingSystem 8.0 ATTACHMENTS 8.1AAMT-M-1-0, Thermal Extractionof Fluorideby Means of a Modified Dohrinann DX2000 OrganicHalideAnalyzer-Liver 8.2 AMDT-M-2-0, FluorideMeasurement by Means of an Orion EA940 Expandable Ion Analyzer 8.3AMDT-M4-0, Extractioonf Fluorochemicalsfrom Rabbit Liver 8.4AAlDT-M-5-0, Analysisof RabbitLiverExtractforFluorochemicalsUsing ElectrosprayMass Spectrometry 8.5 AMDT-M-8-0, AnalysisofFluorideUsing the SkalarSegmented Flow Analyzer with Ion SelectiveElectrode 8.6AMDT-M-14-0, Thermal Extractioonf Fluorideby Means of a Modified Dohrmann DX2000 Organic HalideAnalyzer-Serum 4 0()00!31 3M Environmenial Laborstor Method 'MerrnalExtractionof Fluorideby Means of a ModifiedDohrmann DX2000 OrganicHalideAnalyzer- Liver Method IdentificatiNounmber: AMDT-M-1 RevisionNumber: 0 AdoptionDate: RevisionDate: None Author:RichYoungblom Approved by: roupLear OUP Lt /,PZ-3 1 Dafe QualityAssurance Date Software: MS Word 5.1a AffectedDocuments: AMDT-M-2 FluoridMeeasurementby Means ofan OrionEA940 Exr)andablIeonAnalyzer AMDT-EP-3 RoutineMaintenanceofa ModifiedDohrmann DX2000 OrganicHalideAnalyzer 0()0052 1.0 SCOPE, APPLICABLE C-OMPOUNDS, AND MATRICES 1.1Scope: Thismethod isfortheoperationofa Dohrmann DX2000 when itisused toextract fluoridferom variousmatricesT.he fluoridiestypicallcyollecteidnTISAB solutiofnoranalysis withan ionselectiveelectrode. 1.2ApplicableCompounds: Fluorochemicalsor otherfluorinatecdompounds. 1.3Matrices:Biologicatlissuesp,articularlliyver. 2.0 KEYWORDS 2.1Fluoridef,luorinee,xtractionp,yrolysisi,onizationi,onselectiveelectrodeD,ohrmann, halide, DX2000, fluorochemicals. 3.0 PRECAUTIONS 3.1Glassware and exhaustgasescan be extremelyhot. 3.2Glasswareisfragileb,rokenglassmay causeinjuries. 3.3 Pressurizedgases,propercompressedgas handlingpracticersequired. 3.4Solventbasedsamplesmay flashm,ay need toallowthem todrydown beforestartinrgun. 3.5Potentiabliohazardsdue tothebiologicamlatrices.Use appropriatpeersonalprotective equipment. 4.0 SUPPLIES AND MATERIALS 4.1Compressed Oxygen, Hydrocarbon free,regulatedto30 PSI. 4.2Compressed Helium, High PurityGrade,regulatedto45 PSI. 4.3Quartzglasssample boatwithTeflonTMtubing,Dohrmann 890-097or equivalent. 4.4Quartzglasscombustiontube,RelianceGlass G-9405-012 or equivalent. 4.5Orion940999 TotalIonicStrengthAdjustmentBuffer(TISAB II)or equivalent. 4.6Sample collectiovnialsH,DPE. 4.7 Milli-QTIw-alter 4.8Polystyrenepipettes. 4.9ActivatedCharcoal,E. Merck 2005 or equivalent. 4.10HamiltonSyringeor equivalent. 4.11Miscellaneouslaboratorgylassware 5.0 EQUIPMENT 5.1Rosemount Dohr-mannDX2000 OrganicHalideAnalyzer,modifiedforfluoridextraction. 5.2IBM compatible386 or 486 computer. 5.3DX2000 softwarev,ersion1.00,modifiedforfluoridextraction. 5.4Excel Spreadsheet,version5.0orgreater 6.0 INTERFERENCES 6.1Samplesizieslimitetdoapproximate1l5y0mg,dependinogn samplemoisturceontentT,his may varyfrom matrixtomatrix. 2 o ()o o 7.0 SAN4PLE HANDLING 7.1Samples arenottobe handledwithbarehands. Fluoridemay leachfrom theskintothe sample. Use forcepsor probetotransfetrissues. 7.2Samples ofliverarecutfrom frozenliverand placedina taredand labeledweigh boat.Use a cleanscalpeland cuttingboard.The cuttinbgoard and scalpelshouldbe cleanedwithwater, methanol,ormethanol-watersolutioanftereach liveriscut. 8.0 CALIBRATION AND STANDARDIZATION 8.1PreparationfCalibratiSotnandards 8.1.T1hestandarrdesquirefdoreachprojecwtilnleedtobeapproprifaotrethaitndividuparloject. Refertoprotocolforthatproject. 8.1.2Typically50-500 ppm FC-95 inmethanolstandardsareused. 8.1.3For rabbitliverstudiesu,sebeefliverasthematrix.Cut a pieceoffrozenbeefliver(100 150mg) and weigh itina labeledand taredweigh boat. 8.2 Calibration- Overview The normal calibratioinsthefluoridceurve(ANMT-M-2). However, ifan optionalspikedliver curve isrequiredtheprocedurelistebdelow isused. 8.2.1A calibratiocnurvefortheDX2000 isgeneratedby spikingsampleswithknown standards and combustingthem usingthesame methods and matrixtypeas thesamplestobe tested. 8.2.2Typicallyt,hreereplicateosfeach standardand fiveconcentratioonfs standardswillbe spiked. 8.2.3Standardcurvewillbe plotteads Mass SpikedF (ug)on thex-axisand StandardMass Recovered F (ug)on they-axis.Generatea regressiocnurveand calculatteheequationfortheline and ther2 value. 8.2.4Mass SpikedF (ug)= (Amount spikedinmL) x (Conc.ofstandardinppm) x (0.6004)* *FC-95 is60.04% F therefor0e.6004 isthefactorused toconvertFC-95 toF 8.2.5StandardMass RecoveredF (ug)= (TISAB volume inmL) x (Orionreadinginppm) 8.3Calibratio-n Procedure 8.3.1StartUp 8.3.1.1Run 2 ormore CleanCycleswhen startinignstrw-neenatchday. More cleancyclesmay be usediftheprevioussamplescontainedhighconcentrationosf fluoride. 8.3.2Blanks 8.3.2.1Preparesample usingthesame methods and typeof matrixas thetestsample. 8.3.2.2For rabbitstudiesu,sebeefliverasthematrix.Prepareatleast3 samplesofbeefliver (100 - 150 mg) forblanks. 8.3.2.3Putsample inDohrmann boat.Combust each sample asdescribedinsection9.0and analyzesample accordingtomethod AMDT-M-2 fortheionselectiveelectrodaenalysis. 3 ()()0054 8.3.2.F4orrabbisttudietsh,emeterreadinfgora blanksampleshouldbe0.03ppm orlower beforeproceedingwiththecalibrationB.um samplesuntilthislimitisreached,or untilinthe judgement of theoperatorthereadingisstablewith respecttohistoricarleadings(previous48 hours). 8.3.2.5For non-rabbitstudiest,heblankreadingsshouldreacha predeterminedionconcentration beforeproceedingwiththecalibration. 8.3.2.6Itmay be necessarytomix approximately50 mg ofcharcoalwiththesample toaid combustion. 8.3.3 Standard Curve 8.3.3.1Weigh outatleast15 matrixsamples (5 standardswith 3 replicateesach)intaredand labeledweigh boats.For rabbitstudiesw,eigh 100-150 mg beefliversamples. Record weightsin studydata.Storethematrixsampleson dry iceor icepacksto keep them frozenuntilused. 8.3.3.2 Placeweighed beefliversample inDohrmann sample boat. 8.3.3.3StartwiththeloweststandardconcentrationU.sing a Hamilton syringe,ejecta fixed quantityof thestandardon or in thematrix.For rabbitstudiesu,se4 uL of standardand ejectiton or inthebeefliver. 8.3.3.4At least3 replicatesshouldbe used forthelowest standardconcentrationm;ore replicates may be used atthediscretioonf theanalyst. 8.3.3.5Combust thesample asdescribedinsection9.3and analyzeaccordingtoAMDT-M-2. 8.3.3.6RLm all15 standards.Ifone replicatiessignificantdliyfferenftrom theothertwo replicatesr,an anothersample forthatstandard.Indicatein datathatthenew replicatreeplacesthe oldreplicataend thatthenew replicatweillbe used to calculatteheregressioncurve. 8.3.3.7When allstandardshave been run,calculattehe r2. r2must be atleast0.95. Ifitisnot at least0.95,consultwith supervisor. 8.3.3.8A new standardcurveshouldbe run when thecombustion tubeor sample matrixis changed. New standardcurvemay alsobe run atthediscretioonf the analyst. 8.4 Storage Conditions for Standards 8.4.1Storagerequirementsforstandardsaredependenton theindividualstandardsused. Typically,standardsarestoredatroom temperatureinplastisccrew topbottles. 8.4.2 New FC-95 standardsshouldbe preparedatleastonce a month. 9.0 PROCEDURES 9.1TypicalOperatingConditions: 9.1.1Combustion tubetemperature= 9500C. 9.1.2Oxygen and Helium flow 50 cc/mj*nute. 9.1.3Vaporization/Dryintgime 240 seconds. 9.1.4Bake time = 300 seconds. 9.2Start Up Procedure: 9.2.1Iftheprogram isnot starteds,tarttheEOX program on thePC. 9.2.2Open the SYSTEM SETUP window. 9.2.3Put the furnacemodule and thecellintheREADY mode. 9.2.4Close the SYSTEM SETUP window. 4 0()0055 9.2.5When theoven has reachedtheREADY temperature,run theCLEAN found in theCELL CHECK menu. 9.2.6 See AMDT-EP-3 fordetailosf theDohrmann software. BOAT program 9.3 Sample ExtractionProcedure: 9.3.1Open the SAMPLE HATCH and placethesample intheBOAT. Itmay be necessaryto mix approximately50 mg of charcoalwiththesample toaidcombustion. Ifthisisdone,charcoal should alsobe rra'xeidnwhile establishintghebaselineand when generatingthestandardcurve. 9.3.2 Close SAMPLE HATCH. 9.3.3 Add appropriatveolume of TISAB solutionor 1:1 TISAB:Milli-QTI"watermixture toa labeledsample collectiovnial.Typically0.6n-LLto 15 m-L areused. For rabbitstudies,use 1.0or 2.0 mL of 1:1 TISAB:Milli-@QTIIwIatermixture. 9.3.4 Placethe vialso thatthetipof theCOMBUSTION TUBE isintheTISAB atleast0.25 inches.Gases releasedduringpyrolysismust bubble throughtheTISAB. 9.3.5Run theEOX-SOLIDS program found intheRUN menu. 9.3.6 When theEOX program isfinishedr,emove thecollectiovnia]from thecombustion tube. 9.3.7 IfundilutedTISAB was used tocollecthesample,add an equalvolume ofMilli-QT"water totheTISAB tomake 1:1 TISAB:Milli-QTM. 9.3.8Rinse theend ofthecombustiontubewithMilli-QTMwaterand wipe with a KJMWIPE to remove any TISAB remainingon thetube. 9.3.9 Open thesample hatchand remove any remaining ash from theboat. Ash can be removed with a cottontippedapplicatoorr vacuumed out. Itmay be necessarytoscrapparticleosffthe bottom with a spatulaor othersimilardevice. A drop of Milli-QTIIwIater may be added to the boattoaidintheClean Cycle. 9.3.10 Close thehatch. 9.3.11 Run the CLEAN BOAT program. 9.3.12 Sample isreadyforanalysisby ionselectiveelectrode(AMDT-M-2). 9.4 Sample Calculations. 9.4.1 Use thestandardcurve to calculattehesample value. 9.4.2Sample Mass Recovered F (ug)= (TISAB vol inmL) x (Orionreadiniizn1212m- intereel2t) (Slope) 10.0 VALIDATION 10.1 QualityControl 10.1.1DailyStartUp Check Samples:Once thestandardcurveisestablisheeda,chday of analysisisstartedby analyzingQC samples. The QC samples areto be the same as thelowest concentrationspikedsamples used togeneratethestandardcurve. Each concentrationmust be done intriplicatuenlessthefirsttwo replicateasrewithin20% of thestandardcurve,thena third replicatiesnot necessary. 10.2 Precisionand Accuracy: See method development analysisand sample analysisin FluorideNotebooks 2,3,and 5. Precisionand accuracyvarieswhen analyzingsamples of different matnces and differenrteferencecompounds. 10.3 Other ValidationParameters: NA 5 0()005c) 11.0 DATA ANALYSIS 11.1 Calculations 11.1.1For thestandarcdurve,useregressioannalysiisnExcel,version5.0orgreater. 11.1.2To calculattehefluoridceontractioinnthesample,seemethod AMDT-M-2. 11.2 Analyzing theData 11.2.1r2must beat least0.95orgreater".Outliersm"ay" be excludediftwo of thethreereplicates arewithin20% ofeachotherand theoutlieirsgreaterthan200% oftheaverageofthosetwo or lessthan50% of theaverageof thosetwo. Any suchoutliersshouldbe pointedoutinthedataand notedintheFinalReportalongwiththereasonitwas consideredan outlier. 12.0 ATTACHMENTS None 13.0 REFERENCES 13.1Rosemount Dohrmann DX2000 OrganicHalideAnalyzerOperator'Msanual (Manual 915- 349,revisionB, December 1993) 1 13.2 AMDT-M-2 FluorideMeasurement by Means of an OrionEA940 Expandable[on Analyzer 13.3AN4DT-EP-3 RoutineMaintenanceofa ModifiedDohrmann DX2000 OrganicHalide Analyzer 14.0 REVISIONS Revision Numbe Reason forChan2e Revision Date 6 0()()057 3M Environmental Laboratory Method FluorideMeasurement by Means ofan Orion EA940 Expandable Ion Analyzer MethodIdentificaNtuimobner: AMDT-M-2 RevisionNumber: 0 AdoptionDate: lo-4-fsRevisionDate: None Author:RichYoungblom Approved By: G r Leader Date wla. QualityAssurance L,0-4,-f Date Software-MS Word 5.la AffectedDocuments: AMDT-M-1 ThermalExtractioonfFluoridbey Means ofa Modified DohrrnannDX2000 OrganicHalideAnalyzer 0()OOGS .1.0SCOPE, APPLICABLE COiVtPOIIXD-S@ AND NIATRICES 1.1SCOPE: This method isforthecalibratioand operationofan OrionEA940 Expandable Ion Analyzer. 1.2APPLICABLE COMPOUNDS: Fluoride. 1.3APPLICABLE MATRICES: LiquidsamplesinanappropriatbeuffersolutionP.referred pH of6.0. 2.0 KEYWORDS 2.1Fluoridef,luorinei,on selectiveelectrode 3.0 PRECAUTIONS 3.1No hazardsidentifiewdiththismethod. 4.0 SUPPLIES AND MATERIALS 4.1Orion940999 TotalIonicStrengthAdjustmentBufferIl(TISABII)or equivalent. 4.2Orion Model 900001 electrodfeillinsgolutio(nAgCl)orequivalent. 4.3Orion940907 100 ppm fluoridsetandardor equivalent. 4.4Milli-QTIwIaIteror equivalent. 4.5Magnetic stirbars. 4.6 Lab tissues. 4.7Sample collectiovnials. 4.8Plastic100 mL volumetricflasks. 4.9Polystyrenepipettes. 4.10Miscellaneouslaboratorgylassware. 5.0 EQUIPMENT 5.1Orion Model EA940 ExpandableIonAnalyzeror equivalent. 5.2OrionModel 960900 SolidStateCombinationFluorideelectrodoer equivalent. 5.3Magnetic StirPlate. 5.4IBM compatible386 or486 computer(onlyneededifusingOrion 3E software). 5.5Orion RS232 interfacceable(onlyneededifusingOrion3E software). 5.6MicrosoftExcel 5.0(ordyneededifusingOrion 3E software). 6.0 INTERFERENCES 6.1Itisrecommended thatthepH be atornear6.0.A 1:1 n-iixtuorfeTISAB and sample/MilliQ TlllwatewrillgenerallybringsampletopH of6.0. 6.2Sample temperaturemay effectfluoridmeeasurement. Itisrecommended thatthesample be atroom temperatureasthestandardswere when themeter was calibrated. 6.3The ratethe samplesarestirreadtshouldbe consistenwtiththeratethestandardswere stirred. 2 0()005!1 6.4Air bubblestrappedunderelectrodecan giveerroneousreadings.Make sureno airistrapped under electrode. 7.0 SAMPLE HANDLING 7.1 No specialhandlingnecessary. 8.0 CALIBRATION AND STANDARDIZATION 8.1 Preparationof CaUbrationStandards 8.1.1Measure50 niLof TISAB IIinto5 100mL plastivcolumetrifclasks. 8.1.2Labeltheflaskass0.05,0.1,0.5,1.0,and 1.5ppm F-,alongwiththedateand yourinitials. 8.1.3Pipett0e.05,0.1,0.5,1.0,and 1.5mL of 100 ppm fluoridsetandardintotheappropriately labeledflasks. 8.1.4Add approximately30 niL ofMilli-QTMwatertoeach flask. 8.1.5Shake theflasksto mix thesolutions. 8.1.6Elimffiataeirbubblesfrom theflasksby tippingtheflaskson theirsidesand rollingtheairin theflasksover theairbubbles. 8.1.7Bringthevolume inthe flasksup tothe 100 mL mark withMilli-QTMwater. 8.1.8Invertand shake theflasksforthefinalmixing. 8.1.9Record standardsin StandardsLog Book. 8.2 Calibration 8.2.1Ifnecessary,remove tapefrom electrod@fillinhgole. 8.2.2Invertprobe towet top seal. 8.2.3Ejecta few dropsof fillinsgolutionfrom bottom ofelectrodetowet lower seal. 8.2.4Filltheelectrodewith fillinsgolution. 8.2.5The meter and theF- electrodearetypicallcyalibratebdy directmeasurement withno blank correctionu,singstandardswith concentrationosf 0.05,0.1,0.5,1.0,and 1.5ppm F-,following themanufacturer'sinstructions. 8.2.6Record the slopein the appropriatelog @ook. 8.2.7Clean theelectrodeby rinsingwithMilli-QTMwaterand wiping the sidesdown withlab tissues. 8.3 Storage Conditions for Standards 8.3.1Calibrationstandardsarestoredatroom temperature, 9.0-PROCEDURES 9.1Calibrationand Nleasurement, Standard method: 9.1.1The sampletobe measured needstobe mixed withTISAB usingtheproportions recommended by theTISAB manufacturer. 9.1.2Placea stirbarinthesample and placethesample on thestirplate. 9.1.3Allow thesample tomix fora few secondsbeforeinsertintgheelectrode.When the electrodeisinsertedm,ake surethereareno airbubblestrappedunder theelectrode. 9.1.4The sample shouldbe thesame temperatureas thecalibratiosntandardsand stirreadtthe same rateas thecalibratiosntandards. 9.1.5When thereadingshave stabilizedr,ecordthereadingintheappropriatelogbook. 3 0()00(;o 9.2CalibratioAnd Measurement,UsingOrion3E Software: 9.2.1Calibration: 9.2.1.1Follow steps8.2.1 to 8.2.4. 9.2.1.2Press Function Key #8 (F8). 9.2.1.3The computer screenwillaskyou to confirm thenumber of standardsto be used, concentratioonf thestandards,and whetheror nota blank isto be includedin thecalibration. Make any necessarychanges totheinformationpresentedand clickon CONTINUE. 9.2.1.4Placetheelectrodein thefirssttandardon thestiprlateand clickon CONTINUE. 9.2.1.5Observe thereadingson thegraphicdisplayon thecomputer. When thereadingshave stabilizedp,ressACCEPT R.EADRNG. 9.2.1.6Repeat step9.2.1.4and 9.2.1.5fortheremaining standards. 9.2.1.7Afterthe finalstandard,thecomputer willdisplaytheslopeof thecurve,as wellas the intercepatnd correlationR.ecord theslope,intercepta,nd correlatioinnthe appropriatleog book and clickon CONTINUE. The calibratiodnataisautomaticallcyopiedtoC:\Orion\Data\Calib.txt. 9.2.2Data Spreadsheet: 9.2.2.1SelecteitherNEW or OPEN from theFILE menu toopen a new or existingspreadsheet to storedatain. 9.2.2.2Record the name of the spreadsheetused in the appropriatelog book. 9.2.3Fluoride Measurement: 9.2.3.1Follow steps9.2.1through 9.2.4 9.2.3.2Enter thename of thesample in theappropriateplaceon thescreen. 9.2.3.3Clickon theNEW SAMPLE button' 9.2.3.4When thereadingshave stabilizecdl,ickon theRECORD buttonand writetheresultinthe appropriatleog book. 10.0 VALIDATION 10.1 QualityControl: 10.2 Precisionand Accuracy 10.3 Other ValidationParameters AccordingtoReference13.2,therangeof detectioins0.02 ppm fluoriduep toa saturatedsolutioonf fluoride. 11.0 DATA ANALYSIS 11.1 CalculationsNonenecessary. 11.2 Analyzing the Data Nonenecessary. 12.0 ATTACHNIENTS None 13.0 REFERENCES 4 13.1Orion Model EA940 Expandable Ion Analyzer InstructioMnanual, Orion Research Incorporated,199 1. 13.2Orion Model 960900 SolidStateCombination FluorideElectrodeInstructioMnanual, Orion Research Incorporated1,991. 14.0 REVI TONS Revision Numbe Reason forChange Revision Date 5 0 ()0 0 c 3M En !ntalLaboratory Method Extractionof Fluorochemicalsfrom Rabbit Livers SOP IdentiricatioNnumber: AMDT-M-4 Revision Number: 0 Adoption Date: RevisionDate: None Author.-Dave Christenson/CynthiWaeber Approved By: 04-0? /7uLp2P2L1-e4a:add@er /0 Dam QualityAssurance -3 /-1.5'Date Software:MS Word, 6.0 AffectedDocuments: M-5, Analysisof RabbitExtractforFluorochemicalsUsing Electrospray Mass Spectroscopy. 0()006.'j 1.0 S(:OPE 1. 1 Scope: This method isfortheextractioonf fluorochemicalfsrom rabbitlivers. Ethylacetateisused toextractfluorochemicalsfrom theliversforanalysisby electrospraymass spectroscopy. 1.2 Applicable Compounds: Fluorc>chemicalosr otherfluorinatecdompounds. 1.3 Matrices: Rabbit Livers. 2.0 KEYWORDS 2.1 Fluorocherr@calsr,abbitliverse,lectrospramyass spectrometer,fluorinated compounds, extraction. 3.0 PRECAUTIONS 3.1 Use gloveswhen handlingtherabbitliverst,heymay containpathogens. 4.0 SUPPLIES AND MATERIALS 4.1 Supplies 4.1.1 Syringe,capableof measuring 100 pL 4.1.2 Eppendorf typeordisposablepipets 4.1.3 Gloves 4.1.4 Plasticgrindingtubes 4.1.5 Plasticentrifugetubes,15 mL 4.1.6 Labels 4.1.7 Nitrogen 4.1.8 Timer 4.1.9 FiltersT,itannylon syringefilter0s.,2 4m. 4.1.10 Analyticaplipets:glassvolumetricpipets. 4.1.11 Disposableplastic3 cc syringes. 4.1.12 Crimp cap autovials. 4.2 Reagents 4.2.1 Aqueous Ammonium Acetate(Aldrich)a,pprox.250 ppm: Preparea 2500 ppm aqueous solutionof ammonium acetateby adding 250 mg ammonium acetatetoa 100 mL volumetricflaskand dilutetovolume with MiUi-Q water.Dilutethissolution1:10 fora 250 ppm solution. 4.2.2 Sodium carbonate/SodiumBicarbonateBuffer(J.T.Baker), (Na@CO,jNaHC03) 0.25M: Weigh 26.5g ofsodium carbonate(Na2C03) and 21.0g ofsodium bicarbonate(NaHC03) intoa I L volumetricflaskand bringtovolume withMiUi-Q water. 4.2.3 Diluteacetonitrisloelutiond,iluteacetonitri1l:e1withMilli-Qwater. 4.2.4 EthylAcetate 4.2.5 Methanol 4.2.6 Ntlli-Qwater 4.2.7 IH,IH,2H,2H -perfluorooctanesulfoanciicd(Aldrich) 4.2.8 FC-95 (3M SpecialtCyhemicalDivision) 0()00(i42 5.0 EOUIPMENT 5.1 Ultra-TurraTx25 Grinderforgrindingliversamples. 5.2 Vortex mixer 5.3 Centrifuge 5.4 Shaker 5.5 AnalyticaElvaporator 6.0 INTERFERENCES 6.1 There areno known interferenceastthistime. 7.0 SAMPLE HANDLING 7.1 The rabbitliversarereceivedfrozen,and must be keptfrozenuntiltheextractioins performed. 8,0 CALIBRATION AND STANDARDIZATION 8.1 Preparation of InternalStandards 8.1.1 Preparean internasltandardof approximately12 ppm 1H,lH,2H,2Hperfluorooctanesulphoniacidto be added to each liversample. 8.1.2 Weigh atleast0.1 g of 1H,1H,2H,2H-perfluorooctanesulphoniaccidintoa 100 mL volumenic flaskR.ecord theactualweighl 8.1.3 Bring itup tovolume withmethanol,thisisthestockstandard. 8.1.4 To a 250 mL volumetricflaska,dd 3 mLs of thestockstandardand bringto volume with 1-.Uli-Qwater.Calculatetheactualconcentratioonf the standard. actualmg perfluorocmne- sulphonicacid X 0.1 L 3 mL 250 mL actualconcentrationp,pm 8.2 Prepare FC-95 Anion Standards 8.2.1 PrepareFC-95 standardsforthe standardcurve. 8.2.2 Weigh approximately100 mg of FC-95 intoa 100 mL volumetricflask. Record theactualweight. 8.2.3 Bring up tovolume withdiluteacetonitrile. 8.2.4 Dilutethesolutionwithdiluteacetoniafl1e:10 fora solutionof approximately100 ppm. Dilutethissolution1:10 withdiluteacetoniuilfeor a solutionofapprox. 10 ppm. 8.2.5 Use the 10 ppm solutiontomake working standardswith valuescloseto 5.0ppm, 1.0ppm and 500 ppb. 8.3 Prepare Beef Liver Homogenate to Use for Standards 8.3. 1 Weigh 40 g ofBovine liverintoa 250 mL Nalgene bottlecontaining 200 mLs Milli-Qwater.Grind toa homogenous solution. 8.3.2 Add I mL ofthesolutiontoa 15 mL centrifugteube.Preparea totalofeight I mL aliquotosf thesolutionin15 mL centrifugteubes.Be suretore- suspend soludon by shakingitbetween aliquots. 0()00(;S 3 S.3.3 SpikesevenoftheIn-laLliquowtisththefollowinagmountsofworking standardsin step9.12 of theprocedure.One I mL aliquotservesas the blank. Working Standard UL Approximate final (ApproximateConc.) concentratioonf FC-95 inliver - Blank 500 ppb 100 0.292 ppm 500 ppb 200 0.584 PPM 500 ppb 300 0.877 ppm 500 ppb 400 1.168 ppm I ppm 500 2.924 ppm 5 ppm 5.848 ppm 5 ppm 300 1 8.772 ppm 8.4 Calculatetheactualvalueof thestandards: uL ofstandard x concentratio(nin12p-m-=) finalconcentratio(nppm) 171 mg liver'I/ ml homogenate ofFC -95 inliver *Average weightof bovineliverinsolutionasdeterminedby weighing I mL homogenates of 40 mg @ver in200 mL of MiUi-Q water.The amount of FC-95 isreportedas equivalentsof FC-95 potassium salt. 8.5 Calibration 8.5.1 Extractthespikedbeefliverhomogenate following9.13to9.23of this method. Use thesestandardsto establishyour curve on themass spectrometer. 8.5.2 Alternativelya,standardcurve may be generatedusing ratiosof responses of theperfluorooctansulfonaatneionand theinternalstandardanionversus concentrationof theperfluorooctanesulfonaatneion. 8.6 Storage Conditions for Standards 8.6.1 New standardasrepreparedwitheachanalysisS.tandardsarestoredin covered plasticentrifugteubesuntiltheanalysison themass spectrometer isperformed. 8.7 Storage Conditions for Standards 8. 7. 1 Beef liverhomogenates may be frozenafterpreparation. 9.0 PROCEDURES 9.1 Obtain frozenliversamples.In spenttissuen,ote thattheliverhas notbeen packaged with othertissues. 9.2 Use a dissectinsgcalpeland cutoffapproximatelyI g of liver. 9.3 Weigh thesample directlyintoa mred plastigcrindingtube. 9.4 Record theliverweight in thestudynote book. 9.5 Put a labelon thevialwith thestudynumber, weight,rabbitID, dateand analyst initials. 0()()0664 9.6 9.7 9.8 9.9 9.10 9.11 9.12 9.13 9.14 9.15 9.16 9.17 9.18 9.19 9.20 9.21 9.22 9.23 9.24 9.25 Add 2.5 mLs water. Grind thesample.Put thegrinderprobe in thesample and grindforabout 2 minutes,untilthesample isa homogeneous solutionwith no largechunks. Rinse theprobe offintothesample with 2.5mLs water usinga pipet. Take thegrinderapartand cleanitwith methanol aftereach sample.Follow AMDT-EP-22. Cap thesample and vortexfor15 seconds. PipetI mL intoa 15 mL centrifugteube.Label thecentrifugteubewiththeidentical informationasthegrindingtube.(SeeAMDT-M-4 Worksheet fordocumenting the remainingsteps.) Spikethe beefliverhomogenates withtheappropriataemount of FC-95 standard as describedin 8.3. Spike thesamples and beefliverhomogenates with 100 uL of internalstandard. Add I mL ofthesodium carbonate/sodiumbicarbonatebufferand I mL ammonium acetate. Using an analyticaplipet,add 5 mL ethylacetate. Cap the sample and vortex20 to 30 seconds. Put them in theshakerfor20 niin. Centrifugefor20 to25 minutes,untilthelayersarewellseparated.Set thepower on thecentrifugeto25. Remove 4 mLs of thetoporganiclayertoa fresh15 niL centrifugetubewith a 5 mL graduatedglasspipet.Transferthelabeltothefreshtube. Blow thesample down on theanalyticaelvaporatorto near drynesswith nitrogen, approximately30 to40 minutes. Bring theremainingsample up in I mL diluteacetonitriwliethan analyticaplipet. Vortex 15 seconds. Transferthesample toa 3 mL syringe.Attacha 0.2gm nylon mesh filtera,nd filter thesample intoa freshcentrifugetubeor a autovialL.abel thetube or vialwith the studynumber and animal number. Cap and hold foranalysisby electrospramyass spectroscopy. Complete AMDT-M-4 worksheetand attachtopage of studynotebook. 10.0 VALIDATION 10. 1 QualityControl - not applicable 10.2 Precisionand Accuracy- not applicable 10.3 Other ValidationParameters- notapplicable 11.0 DATA ANALYSIS 11.1 None 12.0 ATTACHMENTS 12.1 WorksheetAMDT-M-4 13.0 REFERENCES 13.1 ANIDT-EP-22 RoutineMaintenanceofUltra-TurraTx-25 14-0 REVISIONS Revision Numl>er Reason forChange Revision Date ()()00(;75 Study Sample Number set R)2nk T-iver Work.-;heeAtMDT-M-4 C-95 approx0.5ppm actual PPM #W 100111 ?t)(i)iT. 100 ul- 400 uL - FC-95 approx Ippm actual ppm #W - li0fI)II, FC-95 approx.5 ppm actual PPM #W Date and InitialfsorStd. '>t)uOL 100 11T qn)dinvlimb= w'here the orityin;iwlnrkqhee I T'ier FxtrAm*r)nP 7)12rea cnny EinerI mT, nf I.iverSniiitinn Pir@-1-0t0 iiTo.f 17 nnm Intem;tlI.Rt2ndArd VnrtexIi ger Pirk-It mT. of ?ro nnrn ArnrnnniiirAncetqtt, PirietI mT, of 0 2,1NaC0,10 '25M NaH('O. Riiffer Pirk-it rnl.nf PthylAret2t-- vn"x 20-10qt-r Sh;ke,70 rnin r-e-ntrifi7i0p-e25 min Rernnye ;14 rnl-:;Iinvinnftnro;vniclaver Blnw tinwn tn neardryne-,-(-<;O 25 mL) viithN, Add I m- nf 1-1 Art-tonitrile.114,0 TN# Vortex IIs Filterugino a '4rrR-r) ;vrinpe with ;10 ')iimSRI filteirntn;vI @ mi- autogamnic,vi:il T)ntek Tniti:ilq 6 3M Environmental Labgrgtor.1 Method Analysisof Rabbit Liver Extract for Fluorochemicalsusing Electrospray Mass Spectroscopy SOP IdentiricatioNnumber: AMDT-M-5 RevisionNumber: 0 Adoption Date: e--4-@r RevisionDate: None Author:Dave Christenson/CynthWieaber Approved By: @u ro/u,ppL:eLaedaedr Date QualityAssurance Date Software:MS Word, 6.0 AffectedDocuments:M-4, ExtractioonfFluorocherr@caflrsom RabbitLivers 0()OOU!) 1.0 SCOPE 1. 1 Scope: Thismethod isfortheanalysisof extractosfrabbitliveror othertissuesor fluidsforfluorochemicalussingthe electrospramyass spectrometer.The analysis isperformed by singleionmonitoringofFC-95 anion,M/Z= 499,theinternal standardMIZ = 427, and otherappropriatemasses. 1.2 Applicable Compounds: Fluorochemicalsor otherfluorinatecdompounds. 1.3 Matrices:RabbitLivers(samples),Beef Liver(standards)o,thertissuesand fluids. 2.0 KEYWORDS 2.1 Fluorochemicals,fluorinatecdompounds, electrospramyass spectroscopy,mass spectrometerr,abbitlivers. 3.0 PRECAUTIONS 3.1 Use cautionwiththevoltagceablefortheprobe.When thevoltagecableisplugged intotheprobe DO NOT TOUCH THE PROBE, thereisriskof electricaslhock. 3.2 Do notrun thepump above it'scapacityof 4000 psi.Ifpressuregoes over 4000 psi stopand releasepressure.The peak tubingmay be plugged.Troubleshootback to findtheplug and replacetheplugged tubing.See AMDT-EP- 15 3.3 Do not run thepump todryness. 4.0 SUPPLIES AND MATERIAILS 4.1 Supplies 4.1.1 Nitrogegnasregulatetdo140 psi. 4.1.2 Fluofixcolumn orequivalent. 4.1.3 100 uLor250 uL flatdp syringeforsampleinjection. 4.2 Reagents 4.2.1 Diluteacetonitfimloebile phase,diluteacetonitri1l:e1 withMilli-Qwater. 4.2.2 Milli-Qwater,allwaterused inthismethod shouldbe Milli-Qwater. 5.0 EOUIPMENT 5.1 VG Trio2000ElectrosprMaayss Spectrometeorrequivalent. 5.2 ISCO SyringePump 5.3 SpectraphysicAsS300 Autosampler 5.4 100 uL Assembly 5.5 Autovialsor capped centrifugteubes. 6.0 INTERFERENCES 6.1 There areno known interferenceastthistime. 7.0 SAMPLE HANDLING 7.1 Keep theextractedsamples incapped 15 mL centrifugetubesor incapped autovials untilready foranalysis. 0()Ob7O 8.0 CALIBRATION AND STANDARDIZATION 8.1 Preparationof CalibrationStandards 8.1.1 Seven beefliverstandardsand one blankbeefliverarepreparedduringthe extractiopnrocedure(.SeeAMDT-M-4, section8.0) 8.2 Calibration 8.2.1 Run thesevenbeefliverstandardstwice,startinwgith theloweststandard to obtainthestandardcurve. 8.2.2 Typicallyone standardisrun aftereach 5 to7 samples.Choose a standard inthesame rangeofconcentratioansthesamples. 8.3 Storage Conditions for Standards 8.3. 1 Freshstandardsarepreparedwitheach analysisS.tandardsarestoredin coveredplasticcentrifugteubesuntiltheanalysison themass spectometeris perfor-medS.amples and standardsareNOT refrigerated. 8.4 Storage Conditionsfor Beef Liver Homogenates 8.4. 1 Beef liverhomogenatesmay be frozenafterpreparation. 9.0 PROCEDURE 9.1 InitiaSlet-up 9.1.1 Setsoftwareto"Operateon",IonMode ES'. 9.1.2 Record backingpressureintheinstrumentlog. 9.1.3 Fillthesolventcylinderwithmobilephase. 9.1.4 Setthepump to"Run".Settheflowto 1000 uL/rninO.bserves&oplets corr@ngoutofthetipof theprobe.The pressureshouldbe at1700 to 1800 psi. 9.1.5 Check thefusedsilicaattheend oftheprobe.Use an eye piecetocheck for chips.The tipshouldbe flatwithno jaggededges.Ifany chipsarefound cutoffthetipofthesilicwaitha column cuttearnd pullthesilictahroughto theappropriatleength. 9.1.6 Check your nitrogensupply.Turn on thenitrogenT.here shouldbe no nitrogenleakingaroundthetipof theprobe.A finemistshouldbe coming outof thetip. 9.1.7 Carefullgyuidetheprobeintotheopening.Inserittuntilitwon'tgo any furtherC.onnect thevoltagecableto theprobe. 9.1.8 Go tothe"Editor"page,and setIonizatioMnode toES*,and the appropriatmeasses to427 and 499. 9.1.9 Ifitisnotinsingleionmode go to"Option"and setSIR. 9.1.1OS tartAcquisitionA.ssign a filename, MO-DAY-YR + lettetR.ecord itin thelog book. 9.1.11 Run thebeefliversamplesfirstr,unningeach standardtwiceatthe beginningoftherun..Run a QC check by runningone standardafterevery 5 to7 samples. 9.2 Manual Injection 9.2. 1 Draw 150 uL ofsampleintoa syringeI.njecthesampleintotherheodyne injectiopnort.Injecstlowly.RecordthesampleID inthelogbook. 9.2.2 Turn thevtlveto"On". 9.2.3 Wait two minutes,andinjecthenextsample. 9.2.4 Record thescannLJmberforeachsimple inthelogbook. o o o ol@:1 9.3 Using the Autosampler 9.3.1 Set up sample trayA, B, or C. 9.3.2 Record thesamples and theirpositionsin theinstrumentlog book.Up to 17 vialsmay be ineach run. 9.3.3 Set-upthesampler: 9.3.3.1 Push thesample button 9.3.3.2 Setsample loop size= 100 uL 9.3.3.3 Setinject/sampl=e 2 9.3.3.4 SetCycle time = 0 9.3.3.5 Name thefileL:ivers 9.3.3.6 Identifythetrayused 9.3.3.7 Add thesamples to Queue by pressing"Enter" 9.3.3.8 Press"Run" to start 10.0 VALIDATION 10. 1 Quality Control 10. 1.1 Run a standardevery5 to 7 samples.Ifa significancthange( 50%) in peak heightoccursstoptherun.Only thesamples beforethelastacceptable standardwillbe used.The remainingsamples willbe reanalyzed. 10.2 Precisionand Accuracy 10. 2.1 See Method ValidationReportnumber ANOT-M-5.0.V I 10.3 Other ValidationParameters 10.4 RefertoMethod ValidatioRneportNumber ANMT-M-5.0.Vl 11.0 DATA ANALYSIS 11.1 11.2 Caiculations Plotthestandardcurve,using themean of thetwo valuesobtainedforeach standard. 11. 2. 1Read peak heightsor areasforthesamples from theprintoutU.se linear regressiontodeterminethesample concentrations. 11. 2.2 Calculatethemg of FC-95 anion,or otherfluorochemicalin thetotalrabbit liver- mg FC-95 anioninthetotalrabbitliver ma FC-95 anionfrom std.curve gms of liverused foranalysis x Totalmass ofliverg,ms 11. 3 11. 4 Make a resulttsableand enteritin thestudybook. Printa chromatogram foreach sample,withthepeaks labeledwiththesample or standardM. Write thestudynumber on theprintouti,nitiadla,te,and put itinthe study folder.Stapleallchromatograms togetherand number pages. 4 12.0 ATTACHMENTS None 13.0 REEERENCES 13.1 AMDT-EP- 17 14.0 REVISIONS Revision Numbe Rea5on for chang& Revision Date 5 0()0()71.3 3M EnvironmentalLaboratory Method Analysis of Fluoride Using the Skalar Segmented Flow Analyzer With Ion SelectiveElectrode Method IdentiricatioNnumber: AMDT-M-8 Adoption Date: RevisionNumber: 0 RevisionDate: None Author:Deb Wright/ CynthiaWeber Approved By: up Leader Date QualityAssurance Date Software:IBM MS Word, 6.0 AffectedDocuments: AMDT-EP-26, Operationand Maintenanceof theSkalarSegmented Flow Analyzer 000074 ]-.0SCOPE 1. 1 1.2 This method isfortheanalysisforfluoridet,hermallyextractedfrom samples using theDotirmann DX2000 (AMDT-M-1), and collecteidnTISAB foranalysiswithan [on SelectiveElectrode(ISE).The analysisisperformed usingtheSkatar Segmented Flow AnalyzerwithISE. Samples can be tissuess,erum, biologicamlaterialo,r othermaterialsextractedon theDohrmann. 2.0 KEYWORDS 2.1 Skalar,segmented flow,fluoride. 3.0 PRECAUTIONS 3.1 Follow standardlaboratorysafetypractices. 4.0 SUPPLIES AND MATERIALS 4.1 Supplies 4.1.1 Sample cups,4 mL plasticups with caps 4.1.2 Autopipets,oxfordorequivalentwithplastidcps 4.1.3 Polypropylenevolumetricflasks,100 mL 4.1.4 Cartridgecomponents,referto theSkalarMethods forcomponents and part numbers. 4.1.5 Sample prerilterEsv,ergreen 4.2 Reagents 4.2.1 Brij35, 30% S.F.A.S.Detergent 4.2.2 TISAB 11buffersolutionP:urchaseTISAB 11ftom Orion.To I liteorf TISAB IIadd 2.5mL or 100 ppm fluoridseolutionand I mL Brij. 4.2.3 Sampler rinsingsolutionD:iluteTISAB 111:1 withMilli-Qwater. 4.2.4 NitricacidsolufiofnordecontaminationI,N (labgrade):Slowly add 64 nilsconcentratenditriaccid(HN03) to250 mLs of Milli-Qwater.Bring thevolume up toI L withMilli-Qwater. 4.3 Standards 4.3. 1 Stocksolution1,00 ppm F: purchasedfrom Orion. 4.3.2 Intermediatestandard,10 ppm: Dilute10 mLs of stocksolutionto 100 mLs with Milli-Qwater.Use polypropylenevolumetricflasks. 4.3.3 Working standard:Make up thefollowingworking standardsby addingthe volumes of intermediatoerstockstandardindicateodn thetable,using oxfordor pumpmare pipets,to50 mLs ofTISAB and dilutingto 100 mLs with Milli-Qwater. Working Standard 0.015 PPM mLs of StockStandard mLs of IntermediateStandard o.13 0.03 ppm 0.3 0.06 ppm 0.6 0.09 ppm 0.9 .12 ppm 1.2 0.15 ppm 1.5 0.3 ppm 0.6 ppm 0.6 2 0()007!3 1.2 ppm 1.2 1.5 ppm 1.5 5.0 EQUIPMENT 5.1 SkalarSegmented Flow Auto AnalyzerSansPluss ystem equipped with ISE 6.0 INTERFERENCES 6.1 High concentrationosfalkalinitcyh,loridep,hosphate,sulfateor ironcan cause interferences. 7.0 SAMPLE LIANDLING 7.1 Samples should be storedinpolyethylenebottlesS.amples shouldbe analyzed within30 days. 8-0 CALIBRATION AN]D STANDARDIZATION 8.1 Preparation of CalibrationStandards 8.1.1 Preparecalibratiosntandardsasin section4.3. 8.2 Calibration 8. 2. 1 The standardsareanalyzedatthebeginningof therun. 8.3 Storage Conditions for Standards 8.3. 1 Standardsarestoredincapped polypropylenevolumetricflasksN.ew standardsarepreparedata minimum of every sixmonths, or as necessary. 9.0 PRO!CEDURE 9.1 Start Up Procedure 9.1.1 Clamp down thepumpdecks, airbarsand sampler-pump tubing. 9.1.2 Put thefluoridelectrodesin theelectrodechamber. 9.1.3 Tum on (hepower of thesampler,pumps, offsetpotentiometerand headng bath. 9.1.4 Put thereagent-lineisntheappropriatebottles. 9.1.5 Tum on theinterfacec,omputer,displayand printerM.ake turn on the interfacebefore the computer. 9.1.6 Let thesystem stabilizfeorapproximately30 minutes. sure you 9.2 Startinga Run 9.2.1 Createa sample tableby selectinFgILES, TABLE, and CREATE, typein the name of the filea@nd pressENTER. 9.2.2 Printthesample table,insened in thesystem tableby pushing ESC, PRINT, GROUP 1.This willprinttheentirerun. 9.2.3 Dial thesampler setfingtso theappropriatenumber of samples,number of seconds forsample wash, and number of seconds forthesample. 9.2.4 Fillthesample traywith thestandards,samples,washes and driftsr.W and FW/RUNOUT cupson thesamplerdo not need tobe filled. 9.2.5 Set the baseline. 3 9.2.5. [SelectGRAPHICS. REAL TIME. Ifyou cannot getreal-timey,ou may be intheData Handling Panel.Switch to theAnalysisPanel by selectingCONTROL PANEL and pushing F7. 9. 2.5. 2 Use thesmallscrewdriverfortheoffsetpotentiometertosetthe baseline.Adjustthebaselineuntilitisapproximately3/4 inchfrom thebottom of thescreen. 9. 2.5. 3 Check thehigheststandardand adjustthegain,ifnecessary,with theinterfacsecrew #3. 9.2.6 Go toCONTROL PANEL, and toanalysispanel.Deselecttheanalysisthat willnot be run.(Selector deselectanalysisby pressingENTER.) PressTab toreturnto theAnalysisPanel. 9.2.7 Press thespacebarto bringup thelocalmenu. 9.2.8 SelectSTART to startheanalysis. 9.2.9 Type your ID (initialst)h,esample tablewhich you createdunder 9.2.1(or pressENTER forchoices),choose runningwithor withoutthesystem table and selectSTART ANALYSIS. 9.2. IOAfter startintghesoftware,starthesampler.Make surethatthesampleris setto therightnumber of samples and thatthesample/wash/airtimes are OK. 9.2.11 SelectGRAPHICS, REAL TIME toview theprogressof theanalysis. 9.3 Loading and Printing the Data-File 9.3.1 Go toCONTROL PANEL, pressthespacebartobringup thelocalmenu and selectLOAD. SelectAUTOCALCULATION and enterthefilename(or highlighthefiletobe printedand pressENTER). 9.3.2 To view thecalibratiocnurve,go toGRAPHICS, CALIBRATION CURVE. 9.3.3 To printthehighlevelcurve,push PRINT SCREEN. 9.3.4 To printthelow levelscreen,push ESC togetout ofgraphics.Select SETTINGS. Change themax y valuetoapproximately900.Go toCAL CURVE and pressESC, andenter. PressPRINT SCREEN. 9.3.5 Return toSETTINGS and change themax valueback to4095, go toEDIT, pressENTER and PRINT SCREEN toprintsample peaks. 9.3.6 To printtheresultgso toCONTROL PANF-L, SPACEBAR, OUTPUT, OUTPUT. SelectPRINTER fortheEpson or PRN fortheLaser. 9.4 Shutdown 9.4.1 Put allthereagent-lineisnMilli-Qwater. 9.4.2 Let thesystem rinseforapproximately30 minutes. 9.4.3 Afterthesystem has rinsedcompletely,turnoffthesampler,pump and offsetpotentiometerT.urn Off theheatingbathon weekends. Leave liquidin the lines. 9.4.4 Take theelectrodeoutand soak in 100 ppm F overnight. 9.4.5 Release thepump-decks, airbarsand sampler ptimp-tubing. 9.4.6 SelectFILES, pressALT F and selectQUIT toexittheprogram. 9.4.7 On Friday,turnoffthecomputer,displayand interfaceforthe weekend. 10.0 VALIDATION 10.1 Quality Control 10. 1.1Run a standard(mid tohighconcentratione)very 10 samples.Ifa significancthange inpeak heightoccurs,only thesamples beforethelast acceptablestandardwillbe used.The remainingsamples willbe reanalyzed. 4 0()0077 10.2 Precisionand Accuracy 10.2.1SeeMethodValidatiRoenportnumberAMDT-M-8.0.Vl 10.3 Other ValidationParameters 10.4 RefertoMethod ValidatioRneportNumber AMDT-M-8.0.Vi 11.0 DATA 11. 1 11.2 11.3 11.4 ANALYSTS Calculations 11. 1. 1The standardcurve isplottedby the Skalarsoftware. 11. 1.2 Allcalculationasredone by theSkalarsoftware.r2 should be 0.995 or better. Prepare spreadsheetsto summarize data.Includesample volume, weights used etc. Write thestudy number on theprintoutsi,nitiadla,te theprintout,and bind together with allpackage documents and placein thestudyfolder.Make a copy of the summary sheetand mpe intothestudy notebook.Back up alldataand spreadsheets onto studydiskand backup disks. ElectmnicData 11. 4. 1 OLP studiesE:lectronicdataiscopiedonto theStudy floppydiskforeach study,and alsodam iscopied onto a floppydisk thatis storedin thelab. 11. 4.2 OtherstudiesA:lldataiscopied onto a floppydiskthatisstoredin thelab. 12.0 ATTACHN4ENTS None 13.0 REFERENCES 13.1 AMDT-M-1, Thermal ExtractioonfFluorideby Means ofa ModifiedDohrmann DX2000 OrganicHalideAnalyzer-Liver 13.2 SkalarMethods, #335,SkalarMethods Manual 13.3 AMDT-EP-26, Operationand Maintenanceof theSkalarSegmented Flow Analyzer 14.0 REVISTONS Revision Numbe Reason forchange Revision Date 5 0()O()7s 3M Environmental Laboratory Method Thermal Extractionof Fluorideby Means of a ModifiedDohrmann DX2000 Organic HalideAnalyzer- Serum Method IdentificatiNounmber: AMDT-M-14 RevisionNumber: 0 AdoptionDate:10-3-.'lr RevisionDate: None Author RichYoungblom Approved by: OD"-%) ::Ul roup Leader p@L 1 @@01-31 Date QualityAssurance z Date Software:MS Word S.Ia AffectedDocuments: AMDT-M-2 FluoridMeeasurementby Means of an OrionEA940 ExpandableIon Analyzer AMDT-EP-3 RoutineMaintenanceofa ModiriedDohrmann DX2000 Orgar@cHalideAnalyzer 0()O()7i) 1.0 SCOPE. APPLICABLE COMPOUNDS, AND MATRICES 1.1Scope: This method isfortheoperationofa Dohrmann DX2000 when itisused toextract fluoridferom variousmatrices.The fluoridiestypicallcyollecteidnTISAB solutionforanalysis withan ionselectiveelectrode. 1.2ApplicableCompounds: Fluorochemicalosr otherfluorinatecdompounds. 1.3Matrices:Biologicaflluidsp,articularsleyrum. 2.0 KEYWORDS 2.1Fluoridef,luorinee,xtractiopny,rolysisi,onizatioino,nselectiveelectrodeD,ohrmann, halide, DX2000, fluorochemicals. 3.0 PRECAUTIONS 3.1 Glasswareand exhaustgasescan be extremelyhot. 3.2Glasswareisfrag0e,broken glassmay causeinjuries. 3.3 Pressurizedgases,propercompressed gas handlingpracticersequired. 3.4Solventbasedsamplesmay flashm,ay needtoallowthem todry down befores=ing run. 3.5Potentiabliohazardsdue tothebiologicamlatrices.Use appropriatpeersonalprotective equipment. 4.0 SUPPLIES AND MATERIALS 4.1Compressed Oxygen, Hydrocarbon ftee,regulatedto30 PSI. 4.2Compressed Helium, High PurityGrade,regulatedto45 PSI. 4.3Quartzglasssample boatwithTeflonTMtubing,Dohrrnann890-097 or equivalent. 4.4Quartzglasscombustiontube,RelianceGlass G-9405-012 orequivalent. 4.5Orion940999 TotalIonicStrengthAdjustmentBuffer(TISAB Il)or equivalent. 4.6Sample collectiovnialsH,DPE. 4.7NElli-QTMwater 4.8 Polystyrenepipettes. 4.9ActivatedCharcoal,E. Merck 2005 orequivalent. 4.10Hamilton Syringeor equivalent. 4.11Miscellaneouslaboratorgylassware 5.0 EQUIPMENT 5.1Rosemount Dohrmann DX2000 OrganicHalideAnalyzer,modifiedforfluoridextraction. 5.2IBM compatible386 or 486 computer. 5.3DX2000 softwarev,ersion1.00,modifiedforfluorideextraction. 5.4Excel Spreadsheetv,ersion5.0orgreater 6.0 IN'TERFERENCES 6.1 Sample sizeisliniltetdoapproximately100pl. Thismay varyf@om matrixtomatrix. (A)oosO 7.0 SAMPLE HANDLING 7.1Samples aretobe handledwithplastipcipettes.A new pipetteistobe used foreachsample. 8.0 CALIBRATION AND STANDARDIZATION 8.1PreparatioonfCalibratioSntandards 8.1.T1hestandarrdesquirefdoreachprojecwtilnleedtobeappropriaftoerthaitndividuparloject. Refertoprotocolforthatproject. 8.1.2Typically50-500 ppm FC-95 inmethanolstandardsareused. 8.1.3For rabbitserum studiesu,sebeefserum as thematrix. 8.2 Calibration- Overview The normal calibratioinsthefluoridceurve(AMDT-M-2). However, ifan optionalspikedserum curveisrequiredtheprocedurelistebdelow isused. 8.2.1A calibratiocnurve fortheDX2000 isgeneratedby spikingsampleswithknown standards and combustingthem usingthesame methods and matrixtypeasthesamples tobe tested. 8.2.2Typicallyt,hreereplicateosfeach standardand fiveconcentrationosf standardswillbe spiked. 8.2.3Standardcurvewillbe plottedas Mass SpikedF (ug)on thex-axisand StandardMass RecoveredF (ug)on they-axis,Generatea regressiocnurveand calculatteheequationfortheline and ther2 value. 8.2.4Mass SpikedF (ug)= (Amount spikedinmL).x (Conc.ofstandaridnppm) x (0.6004)* *FC-95 is60.04% F therefor0e.6004isthefactorused toconvertFC-95 toF 8.2.5StandardMass RecoveredF (ug)= (TISAB volume inmL) x (Orionreadinginppm) 8.3Calibration-Procedure 8.3.1StartUp 8.3.1.1Run 2 ormore CleanCycleswhen startinignstrumenetachday. More cleancyclesmay be used iftheprevioussamplescontainedhighconcentrationosf fluoride. 8.3.2Blanks 8.3.2.1Preparesample usingthesame methods and typeof matrixas thetestsample. 8.3.2.2For rabbitstudiesu,se beefserum as thematrix. 8.3.2.3Putserum blanji,n.Dohrrnannboat.Combust sample asdescribedinsection9.0and analyzesample accordingtomethod AMDT-M-2 fortheionselectiveelectrodaenalysis. 8.3.2.4For rabbitstudiest,hemeterreadingfora blanksampleshouldbe 0.03ppm or lower beforeproceedingwiththecalibra6on.Bum samplesuntilthislimitisreached,or untilinthe judgement oftheoperatorthereadingisstablewithrespecttohistoricarleadings(previous48 hours). 8.3.2.5For non-rabbitstudiest,heblankreadingsshouldreacha predeten-nineidonconcentration beforeproceeding%kll'tthhecalibration. 8.3-2.6Itmay be necessarytomix approximately50 mg of charcoalwiththesample toaid combustion. 3 0()O()Nl 8.3.3 Standard CurN,e 8.3.3.1Ifbeef serum isfrozen,thaw at leastenough tocomplete thestandardcurve analysisforthe day (=30 mL). 8.3.3.2 PipetteI00@LL of beef serurnintoDohnnann sample boat. 8.3.3.3Startwith theloweststandardconcentrationU.sing a Hamilton syringe,ejecta fixed quantityofthestandardon orinthematrix.For rabbitstudiesu,se 4 uL of standardand ejectiton or inthebeef serum. 8.3.3.4At least3 replicateshouldbe used forthelowest standardconcentrationm;ore replicates may be used atthediscretionof theanalyst. 8.3.3.5Combust thesample as describedin section9.3and analyzeaccording to AMDT-M-2. 8.3.3.6Run all15 standards.Ifone replicatiessignificantdliyfferenftrom theothertwo replicatesr,un anothersample forthatstandard.Indicatein datathatthenew replicatreeplacesthe oldreplicataend thatthenew replicatweillbe used to calculatteheregressioncurve. 8.3.3.7When allstandardshave been nm, calculattehe r2. r2 must be atleast0.95. Ifitisnot at least0.95,consultwith supervisor. 8.3.3.8A new standardcurve shouldbe run when thecombustion tubeor sample matrixis changed. New standardcurvemay alsobe run atthediscretioonf theanalyst. 8.4 StorageConditionsforStandards 8.4.1Storagerequirementsforstandardsaredependent on theindividualstandardsused. Typically,standardsarestoredatroom temperatureinplasticscrew topbottles. 8.4.2 New FC-95 standardsshould be preparedat leastonce a month. 9.0 PROCEDURES 9.1TypicalOperating Conditions: 9.1.1Combustion tubetemperature= 950'C. 9.1.2Oxygen and Helium flow= 50 cc/rninute. 9.1.3Vaporization/]Dryintgime = 240 seconds. 9.1.4Bake time = 300 seconds. 9.2Start Up Procedure: 9.2.1Iftheprogram isnot starteds,tarttheEOX program on thePC. 9.2.2Open the SYSTEM SETUP window. 9.2.3Put thefurnacemodule and thecellinthePEADY mode. 9.2.4Close the SYSTEM SETUP window. 9.2.5When theoven has reachedtheR.EADY temperature,run theCLEAN BOAT program found in theCELL CHECK menu. 9.2.6 See AMDT-EP-3 fordetailsofthe Dohrmann software. 9.3Sample ExtractionProcedure: 9.3.1Open the SAMPLE HATCH and pipetteIOOGL of sample intotheBOAT. Itmay be necessarytomix approximately50 mg ofcharcoalwiththesample toaidcombustion. Ifthisis done,charcoalshouldalsobe mixed in whileestablishintghebaselineand when generatingthe standardcurve. 9.3.2 Close SA%IPLE HATCH. 4 9.3.3Add appropriatveolume ofTISAB solutionor 1:1 TISAB:Milli-QTM watermixturetoa labeledsample collectiovnial.Typically0.6mL to 15 mL areused. For rabbitstudiesu,se 1.0or 2.0 mL of 1:1 TISAB:Milll-QTM watermixture. 9.3.4 Place thevialso thatthetipof theCOMBUSTION TUBE isintheTISAB atleast0.25 inches.Gases releasedduringpyrolysismust bubble throughtheTISAB. 9.3.5Run the EOX-WATER program foundinthe RUN menu. 9.3.6 When the EOX program isfinishedr,emove thecollectiovnialfrom thecombustion tube. 9.3.7 IfundilutedTISAB was usedtocollecthesample,add an equalvolume of Milli-QTMwater totheTISAB tomake 1:1 TISAB:Milli-QTM. 9.3.8Rinse theend ofthe combustiontubewithMilli-QTMwaterand wipe with a KIMWIPE to remove any TISAB remainingon thetube. 9.3.9 Open thesample hatchand remove any remaining ash from theboat. Ash can be removed witha cottondpped applicatoarnd/orvacuumed out. Itmay be necessarytoscrapparticleosffthe bottom witha spanilaorothersirr@ladrevice.A drop of Milli-QTMwater may be added tothe boatto aidintheClean Cycle. 9.3.10 Close thehatch. 9.3.11 Run the CLEAN BOAT program. 9.3.12Sample isreadyforanalysibsy ion selectiveelectrode(AMDT-M-2). 9.4 Sample Calculations 9.4.1Use the standardcurvetocalculattehesample value. 9.4.2Sample Mass RecoveredF (ug)= (TISAB volinmL) x (Orionreadinein122m - intercel2t) (Slope) 10.0 VALIDATION 10.1QualityControl 10.1.D1ailyStartUp CheckSamples:Oncethestandarcdurveisestablisheeadc,hdayof analysisisstartedby analyzingQC samples. The QC samples aretobe the same as the lowest concentrationspikedsamples used togeneratethestandardcurve.Each concentrationmust be done in triplicatuenlessthefirsttwo replicateasrewithin20% of thestandardcurve,then a third replicatiesnot necessary. 10.2 Precisionand Accuracy: See method development analysisand sample analysisin FluorideNotebooks 2,3,and 5. Precisionand accuracyvarieswhen analyzingsamples of different matricesand differenrteferencecompounds. 10.3 Other ValidationParameters: NA 11.0 DATA ANALYSIS 11.1 Calculations 11.1.1 For thestandardcur-,,ues,e regressionanalysisin Excel,version5.0or greater. 11.1.2To calculatethefluoridceontractioinnthe sample,seemethod AMDT-M-2. 5 11.2AnalyzingtheData 11.2.1r2mustbeatleas0t.95orgreate'r*.Outliemrasy" beexcludeidftwoofthethrereeplicates arewithin20% of each otherand theoutlieirsgreaterthan 200% of theaverage of thosetwo or lessthan50% oftheaverageof thosetwo. Any such outliersshouldbe pointedout inthedataand noted inthe FinalReport alongwith thereasonitwas consideredan outlier. 12.0 ATTACHMENTS None 13.0 REFERENCES 13.1Rosemount Dohrmann DX2000 OrganicHalide AnalyzerOperator'sManual (Manual 915349,revisionB, December 1993) 13.2 AMDT-M-2 FluorideMeasurement by Means of an Orion EA940 Expandable Ion Analyzer 13.3AMDT-EP-3 Routine Maintenanceof a Modified Dohrmann DX2000 OrganicHalide Analyzer 14.0 REVISIONS Revision Number Reason forChanize Revision Date 6 9.13 Amendment to analyticaplrotocolAMDT-01 1095.1 I 0()()OSS AnachmentI GLP Study ProtocoAlmendment StudyNumber:AMDT-0 11095.1 StudyTitle: SingleDose DerrnalAbsoption/ToxiciotfyT-60676,T-6068 and T-6069 inRabbits StudyDirector:James D. Johnson Amendment Date: November 10,1995 Amendment Number: I Thisamendment modifiesthefollowingportionof theprotocol: AMDT-M- 14-0specifieussingbovineserum forblanksand QC check samplesin thethermalextractionH.owever, rabbitservm from thecontrolanimals (ANMT- 110394.1)was used was used becausethebovineserum blankswere higher than thesamples. Approved by: tudUylr c@ bate 7 0()O(),L-lc 9.3 QualityAssurance Unit Statement 0()0()87 AttachmentD GLP Study QualityAssurance Statement Study Title: Single-dose Dermal Absorption/Toxicity Study of T-6067, T-6068 and T-6069 in Rabbits Study Number. AMDT-0 11095.1 Name of Auditor:KariRambo Ibis studyhas been inspectedby theQualityAssurance Unit asindicatedinthefollowingtable. The findingswere reportedto thestudydirectoarnd management. InspectionDates EMM T-Q 11-15-95 11-20-95 Phase FinalReport Date InspectionReported to Management StudyDirect 11-20-95 11-20-95 QAU Auditor Date 9.4 Key PersonnelInvolved in the Study 3M Environmental Laboratory Key Personnel Thermal extractiofnollowebdy analysiussingOrionionanalyzer: Jim Johnson Deb Wright Rich Youngblom Deann Plummer Analysisofliverextractsusingelectrospramyass spectrometry: Jim Johnson Dave Christenson Thermal extractionfollowedby analysisusingSkalar segmented flow analyzerwith ionselectiveelectrode: Jim Johnson Deb Wright Rich Youngblom Deann Plummer Documentation and Reporting: Jim Johnson Rich Youngblom QualityAssurance Unit: Gale Van Buskirk CynthiaWeber KariRambo 0()009() 9.11 Data 0()O(.)gl 9.11.1Summary and raw data;ug F' inwhole liveras determinedby thermalextractiofnollowedby analysisusing Orion ion analyzer. 0 ()()()!)O''WLI Summary ofCombustion Data - Liver AMDT-01 1095.1,HWI 6329-152 As Referenced inFinalReport section6.0DATA ANALYSIS Totalug FluorideinWhole Liver Mean per Dose Group* gg Std.Dev. ControlGroup 12.5 + 2.1 4.0mg/kg dose (T6067) 12.5 + 1.6 16 mglkg dose (T6067) 14.7 + 4.0 40 mg/kg dose (T6067) 16.5 + i.2 Fabricexposure(T6068)** 15.7 + 4.2 Fabricexposure(T6069)** 12.8 + 3.7 *Calculatedas the mean oftriplicatseamples from each ofthreemale and threefemale rabbits. **Administered as a 10 cm 1 10 cm pieceof testmaterial(fabric). RPTI 52L.XLS Li3492 AB ID liverblank-1 liverspk-I liverspk-2 liverspk-3 liverspk-4 liverspk-5 liverspk-6 liverblank-2 F5271 1-1 F52711-2 F52711-3 F5271 9-1 F52719-2 F52719-3 Liverblk1 Liverbik2 Liverspk 1 Liverspk 2 F52725-1 F52725-2 F52725-3 F52713-1 F52713-2 F52713-3 F5271 8-1 F52718-2 F52718-3 F52723-1 F52723-2 F52723-3 F52721-1 F52721-2 F52721-3 F52727-1 F52727-2 F52727-3 F52806-1 F52806-2 F52806-3 F52715-1 F52715-2 F52715-3 F52738-1 F52738-2 F52738-3 % rcvry 97% 85% 85% 107% 97% 95% 97% 98% Actual ppm Fin liver (WAN) 0.213 1.10 0.872 0.883 2.63 2.00 2.55 0.159 0.167 0.144 0.149 0.166 0.157 0.205 0.250 0.169 1.19 1.37 0.228 0.231 0.230 0.172 0.152 0.170 0.161 0.156 0.233 0.175 0.230 0.155 0.218 0.191 0.142 o@194 0.206 0.160 0.158 0.155 0.166 0.226 0.255 0.211 0.178 0.169 0.166 Average ppm Finliver (W/W) 0.153 0.176 0.230 0.164 0.183 0.187 0.184 0.187 0.160 0.230 0.171 Liver burned (grams) 0.125 0.134 0.147 0.147 0.123 0.147 0.112 0.140 0.134 0.141 0.149 0.137 0.124 0.133 0.129 0.142 0.124 0.109 0.126 0.116 0.124 0.154 0.166 0.163 9.128 0.135 0.126 0.171 0.150 0.156 0.101 0.116 0.154 0.153 0,136 0.134 0.134 0.137 0.125 0.127 0.133 0.122 0.134 0.126 0.131 Whole liver weight (grams) 66.1 66.1 86.1 76.5 76.5 76.5 69.5 69.5 69.5 71.4 71.4 71.4 57.7 57.7 57.7 72.6 72.6 72.8 62.9 62.9 62.9 69.2 69.2 69.2 69.8 69.8 69.8 62.1 62.1 62.1 64.6 64.6 64.6 Total Finwhole liver (4g) 10.1 13.5 16.0 11.7 10.6 13.6 11.6 12.9 11.1 14.3 11.0 Dosage (mg/kg) 0.0 0.0 0.0 4.0 4.0 4.0 0.0 0.0 0.0 4.0 4.0 Page I oo009.,t RPTI 52L.XLS Ll 3492 AB ID F52809-1 F52809-2 F52809-3 F52724-1 F52724-2 F52724-3 F52737-1 F52737-2 F52737-3 liverbink-I liverspk-1 liverspk-2 liverspk-3 F52801-1 F52801-2 F52801-3 F52720-1 F52720-2 F52720-3 F52733-1 F52733-2 F52733-3 F52788-1 F52788-2 F52788-3 Liverbik1 Liverbik2 Liverspk 1 Liverspk 2 F52730-1 F52730-2 F52730-3 F52731-1 F52731-2 F52731-3 F52802-1 F52802-2 F52802-3 F52714-2 F52714-1 F52714-3 F52734-1 F52734-2 F52734-3 F52774-1 F52774-2 F52774-3 % rcvry 89% 86% 91% 111% 108% Actual ppm Fin liver (WfW) 0.432 0.159 0.168 0.187 0.235 0.192 0.163 0.145 0.155 0.344 1.10 0.973 0,924 0.372 0.278 0.161 0.252 0.206 0.225 0.224 0.182 0.208 0.179 0.154 0.165 0.760 0.301 1.06 1 @02 0.339 0.237 0.380 0.163 0.202 0.160 0.286 0.542 0.244 0.247 0.454 0.281 0.223 0.297 0.183 0.287 0.215 0.174 Average ppm Finliver (WNW) 0.253 0.204 0.154 0.271 0.228 0.205 0.166 0.318 0.175 0.357 0.327 0.234 0.226 Liver burned (grams) 0.138 0.128 0.112 0.119 0.139 0.151 0.129 0.147 0.142 0.134 0.123 0.134 0.149 0.112 0.119 0.148 0.128 0.133 0.130 0.123 0.139 0.139 0.143 0.146 0.121 0.140 0.116 0.160 0.160 0,123 0.113 0.159 0.158 0,108 0.158 0.150 0.110 0.122 0.154 0.152 0.151 0.111 0.146 0.143 0.138 0.122 0.137 Whole liver weight (grarns) 55.4 55.4 55.4 82.0 82.0 82.0 62.5 62.5 62.5 76.7 76.7 76.7 68.7 68.7 68.7 67.7 67.7 67.7 68.0 68.0 68.0 57.9 57.9 57.9 59.0 59.0 59.0 71.9 71.9 71.9 45.5 45.5 45.5 58.1 58,1 58.1 71.1 71.1 71.1 TotalFinwhole liver (gg) 14.0 16.8 9.6 20.7 15.7 13.9 11.3 18.4 10.3 25.7 14.9 13.6 16.0 Dosage (mg/kg) 4.0 16.0 16.0 16.0 16.0 16.0 16.0 40.0 40.0 40.0 40.0 40.0 40.0 Page 2 RPTL 52L.XLS Li3492 AB ID F52712-1 F52712-2 F52712-3 F52729-1 F52729-2 F52729-3 F52803-1 F52803-2 F52803-3 F52716-1 F52716-2 F52716-3 F52728-1 F52728-2 F52728-3 F52787-1 F52787-2 F52787-3 Liverblk1 Liverbik2 Liverstnd 1 Liverstnd 2 52717-1 52717-2 52717-3 52735-1 52735-2 52735-3 52736-1 52736-2 52736-3 52726-1 52726-2 52726-3 52732-1 52732-2 52732-3 52739-1 52739-2 52739-3 Liverspk-3 Liverspk-4 Liverspk-5 Liverspk-6 Liverspk-7 Liverspk-8 Liverspk-9 Actual ppm F- % inliver rcvry (WM) 0.442 0.336 0.210 0.291 0.185 0.184 0.294 0.193 0.148 0.179 0.165 0.177 0.192 0.166 0.215 0.177 0.242 0.190 0.255 0.250 87% 1.14 88% 1.13 0,360 0.202 0,265 0.206 0,203 0.191 0.158 0.168 0.155 0.135 0.143 0.173 0.162 0.148 0.163 0.313 0.127 0.133 0.69 0.923 0.8222 0.888 0.8295 1.11 1,039 1.23 0.769 2.29 0.8718 1.82 0.9319 2.35 Average ppm Fin liver (WM) 0.330 0.220 0.212 0.173 0.191 0,203 0.276 0.200 0.160 0.150 0.158 .0.191 Liver burned Whole liver weight (grams) 0.115 0,134 0.111 0.127 0.135 0.132 0.118 0.115 0,140 0.117 0.137 0.121 0.125 0.126 0.106 0.141 0.110 0.128 0.147 0.122 0.115 0.118 0.119 0.121 0.122 0.137 0.129 0.111 0.160 0.117 0.145 0.152 0.165 0.113 0.156 0.130 0.113 0.127 0.162 0.160 0.113 0.140 0.113 0.128 0.102 0.145 0.120 (grams) 72.0 72.0 72.0 63.1 63.1 63.1 71.1 71.1 71.1 67.3 67.3 67.3 79.9 79.9 79.9 71.2 71.2 71.2 73.0 73.0 73.0 53.9 53.9 53.9 72.9 72.9 72.9 74.4 74.4 74.4 66.1 66.1 66.1 64.9 64.9 64.9 TotalFinwhole liver (lig) 23.7 13.9 15.1 11.7 15.3 14.5 20.1 10.8 11.7 11.2 10.4 12.4 Dosage (mg/kg) Fabric Fabric Fabric Fabric Fabric Fabric Fabric Fabric Fabric Fabric Fabric Fabric Page 3 9.11.2Summary and raw data;analysisof liverextracts usingelectrospramyass spectrometry. This data,althoughsupportivei,ntheopinionof the Study Directorisnot requiredtoreachthe conclusionstatedin FinalReport Section6.0,and thereforeisnot discussedin detail. 0()0097 HVVI* 6329 -152A D'i@e A-i Study: Protocol Number: Test Material: Matrix: R Squared Value: Response Factor Anmunt: Analyst: Data Analyzed: Method: lnsvunwnt: LABOASE FILE SingleDose Dermal Absorption TP3016.AB T-6o67, T-6068.and T-6069 inRabbits(Ll3492) Liver Screening N/A DLC 4/25/95 Fisons VG 2000 ElectrosprayMS 0425950 OW 0 Y, Corr .0- W;4,L, Pa.L." cLL -@-c kitSita6-. 4o,.aaj,-@r. 0@ 00 caos v- c-,.a. AAE LEQ.S 6 TEr--n -QLC- "/1-5/t,5 ook-' --t Group Dose Group 1: DistilleWdater 2-0 mL/kg Group 2: 4 nV/kg T-6067 Group 3: 16 rngtkg T-6067 Group 4: 40 nV/kg T-6067 Group 5. Fabric T-6068 Sample a F52725 F52711 Ion Count Area N/D N/D F-xtractedwt 9 Dilution factor Concentration go/g Totalmass of liver 9 Totalamount of FC-95 per liver mg N/D N/D MID N/D F52715 NID FSZ718 NIO F52723 N/D N/D N/D MD N/D N/D N/D F52724 N/D F52637 N/D F5Z788 N/D F52733 N/D N/D N/D N/D N/D N/D N/D WD N/D F52774 N/D F52731 N/D F52734 N/D F52714 N/D F52802 N/D F52728 N/D F52729 N/D F52716 NIO F52712 NIO N/D N/D NIO N/D N/D N/D N/D N/D N/D MID N/D N/D N/D N/D N/D N/D N/D N/D Group 6: Fabric T-6066 F52739 N/D F52736 NtD F52726 N/D N/D N/D N/D N/D wo wo SIR of K 412 413 The concentrationwas calculatedby using the standard curve and multiplyingthe resultby 415.The 4/S factorIstheresultofa rrow=lculadon inapplying formula 8.4InMethod AMDT-0&4-0. 137 mg of liverwas used In thiscalculationratherthen 171 mg. The concentrationsIn thestandard curve are therefor*5/4largerthen they should be. By multiplyingthecalculatedconcentrationInthe standard curve by 415.thecorrectresultIsobtairwd. 0()00!)S l@HWI#6329-151-&--152; CMPD 042595D Sm (Mn, 2x3);Sm (Mn,2x3) 100 2110 286 435 1223 1377 1052 5 4 759 894 % 1860 1657 1678 2458 2 2296 0- 200 '460.....6.6.0 800 1000 1200..1.'141001-.'.1..6@00- 1800-------- --0--- 2400 ...2.6... HWI # 6329-151 & 152; CMPD L13492 042595D Sm (Mn. 2x3) ioo 2109 285 434 1222 1377 1052 563 758 894 % 1657 1 60 1679 2458 26 2296 042595D Sm (Mn, 2x3) 100 251 344 46 82 145 734 864 589 502 651 1198 1050 1354. 1402 1514 18521911 1695 2432,2460 2303 213;0 % D- 26'0......46'0... . . .. . . 1000 1200 140''0'' 1600 1800 200'0 22'00' 2400 260 -iI HWI # 6329-16 1 & 152; CM PD Li3492 'T I 042595D Sm (Mn, 2x3);Sm (Mn, 2x3) 2110 100 2 6 435 1223 1377 1052 564 79 84 1657 1860 % 1678 2458 2296 26 042595D Sm (Mn, 2x3) 100 574 737 861 1224 898 1196 1380 649 250 4@o 501 1052 983 1 107 1696 1650, 2430 1865 1881 2107 2303 2197 2 25@22520, 9.11.3Summary and raw data; ppm F-in serum as determinedby thermalextractiofnollowedby analysisusing Skalarsegmented flow analyzerwith ion selectivelectrode. 0()0100't,41 6A).4J 'IZS RE: 6329-152 SERUM SAMPLES AMDT 11095.1 Date of Analysis:8/22,8/23,and 8/28/95 Analyst:DDW The samples are burned inthe Dohrman at 950 C using 0.10 mL ofthe serum. The gas iscollectedin2.0 mL of 1:1TISAB/Milli-Qwater. The samples are then analyzed on a Skalar Segmented Flow Analyzer using the Ion SpecificElectrode (ISE)Method. TISAB bufferisadded to each sample as itproceeds through the system. The sample then goes through a heated mixingcoilbeforethe potentialbetween the ion selectiveelectrodeand the referenceelectrodeismeasured. The signalis amplifiedand relatedto thefluorideconcentration. The instrumentwas calibratedinthe ranges of 0.015 - 0.15 ppm and 0.15 - 1.50 ppm fluoride.The standardcurve forthe high range was plottedusing the inverselogarithmoption.The standard curve forthe low range islinear.Allstandardsand samples were then calculatedby the Skalar software using these curves.Allresultsbelow 0.0001 ppm appear on the raw data as A qualitycontrolstandardwas analyzed every 10 samples to check foraccuracy and drift. Raw data istaken from the appropriatecalibratedrange ofthe Skalar printout and summarized on an Excelspreadsheet. The finalresultsare adjustedforthe collectionvolume and any subsequent dilutions. ooolO.3 /I ky .I .. --- ------------------.-.-.-.-.--.-..-..-..-..-..-.-.-.-.--.-.--.-- SUMMARY OF 6329-152 SERUM SAMEPLES AMDT 11095.1 GROUP I Dose Level . 0 mg&g F5271 1-1 F52719-1 F52725-1 F52721-1 F52727-1 F52806-1 0.84 0.68 0.66 0.51 0.48 0.53 0.42 0.49* 0.45 0.34 0.73 ND 0.31 0.96* 0.79 ND 0.84* 0.63 0.60 0.82* 0.53 0.62 0.92 0.49 GROUP2 Dose Level :4 mg/kg F52713-1 F52718 F52723-1 F52715-1 F52739-1 F52909-1 1.36 0.49 0.64 0.31 1.16 0.42 0.99 0.33 0.93 0.52 1.25 0.32 1.00 0.54 0.69 0.49 1.17 0.43 1.40 0.38 1.49 0.61 0.72 0.33 GROUP3 Dose Level - 16 mg/kg F52724-1 F52737-1 F52801-1 F52720-2 F52733-1 F52789-1 1.21 1.17 1.05 1.11 0.32 0.52 0.64 0.74 0.48 0.64 0.72 0.92 1.20 l@29 0.73 0.92 0.92 0.64 O@70 0.54 0.74 0.50 ND GROUP4 Dose Level: 40 mglkg F52730-1 F52731-1 F52802-1 F52714-1 F52734-1 F52774-1 1.04 0.95 1.11 0.97 1.07 1.23 0.59 0.03 0.68 0.95 0.69 0.72 O@39 D@39 0.59 0.93 1.35 0,92 GROUPS Dose Level: 10 110 fabric F52712-1 F52729-1 F52803-1 F52716-1 F52728-1 F52787-1 1.28 0.95 0.69 1.080 0.62 0.67 1.31 0.63 0.92 1.39 0.65 0.77 1.44 0.77 0.68 1.18 0.68 0.76 GROUP6 Dose Level :10 1 10 fabric F52717-1 F52735-1 F52736-1 F52726-1 F52732-1 F52739-1 1.71 t@49 1.16 0.94 1.33 0.90 0.62 0.74 0.57 0.790 o@69 0.59 1.03 0.67 0.60* 0.54 0.42 0.33 Avmgc ofdupliratienaivsistakenforfinalresult-DDW IS2S-SUM.XLS Pop I ooolo,,l SERUM CURVE 2 7-27-95 CARLTON k L,)m 1 10 Is Sample ID Spk 34-1 Spk 34-2 Spk 34-3 Spk 40-1 Spk 40-2 Spk 40-3 Spk 62-1 Spk 62-2 Spk 62-3 Spk 100-1 Spk 100-2 Spk 100-3 Spk 124-1 Spk 124-2 Spk 124-3 Spk 250-1 Spk 250-2 Spk 250-3 Spk 500-1 Spk 500-2 Spk 500-3 Skalar Result (ppm) DI:TISAB mL FC 95 Cone finalvol Solution FC 95 Soln (mL) Spiked (ppm) Mass Spiked (ugF-) Avtrage % Mass Recovery Recovered (ugF-) 0.07 0.06 0.06 2.0 0.004 34.00 2.0 0.004 34.00 0.08 0.12 151% 2.0 0.004 34.00 0.06 2.0 0.004 40.00 0.06 2.0 0.004 40.00 0.10 0.13 132% 0.07 2.0 0.004 40.00 0.08 2.0 0.004 62.00 0.08 2.0 O@004 62.00 0.15 0.17 114% 0.09 2.0 0.004 62.00 0.10 2.0 0.004 100.0 0.11 2.0 0.004 100.0 O@24 0.22 93% 0.12 2.0 0.004 100.0 0.12 2.0 0.004 124.0 0.14 2.0 0.004 124.0 0.30 0.28 94% 0.15 2.0 -.0.004 124.0 0.33 2.0 0.004 250.0 0.23 2.0 0.004 250.0 0.60 0.52 87% 0.23 2.0 0.004 250.0 0.47 0.41 0.50 2.0 0.004 500.0 2.0 0.004 '500.0 1.20 0.92 77% 2.0 0.004 500.0 SERUM CURVE I (CARLTON) 7-27-95 100. ,a 0.90- 1 04 o@so 070 > 060 - 0,50 0.40 0.30 w > 010 o@oo 0.00 0'.10 040 0.60 o.so 100 1.20 1 %iAm SPEKM (mg) SERCRV2C.RPT Page 1 0()01()!-3 SERUM CURVE 2 7-27-95 CARLTON Sample ID Spk 34-1 Spk 34-2 Spk 34-3 Spk 40-1 Spk 40-2 Spk 40-3 Spk 62-1 Spk 62-2 Spk 62-3 Spk 100-1 Spk 100-2 Spk 100-3 Spk 124-1 Spk 124-2 Spk 124-3 Spk 250-1 Spk 250-2 Spk 250-3 Spk 500-1 Spk 500-2 Spk 500-3 Skalar Result (ppm) DI:TISAB mL FC 95 Conc finalvol Solution FC 95 Soin (mL) Spiked (ppm) Mass Spiked (ugF-) Mass % Recovered Recovm (ugF-) 0.07 2.0 0.004 34.00 0.08 0.14 171% 0.06 2.0 0.004 34.00 0.08 0.11 138% 0.06 2.0 O@004 34.00 0.09 0.12 145% 0.06 2.0 0.004 40,00 0.10 0.12 123% 0.06 2.0 0.004 40.00 0.10 0.12 128% 0.07 2.0 0.004 40.00 0.10 0.14 1"% 0.09 2.0 0.004 62.00 0.15 0,16 110% 0.08 2.0 0.004 62.00 0.15 0.16 106% o@09 2.0 0.004 62.00 0.15 0.19 126% 0.10 2.0 0.004 100.0 0.24 0.20 85% 0.11 2.0 0.004 100.0 0.24 0.23 96% 0.12 2.0 0.004 100.0 0.24 0.23 98% 0.12 2.0 0.004 124.0 0.30 0.25 84% 0.14 2.0 0.004 124.0 0.30 0.29 97% 0.15 2.0 0.004 124.0 0.30 0.31 103% 0.33 2.0 0.004 250.0 0.60 0.66 109% 0.23 2.0 0.004 250.0 0.60 0.45 76% 0.23 2.0 0.004 250.0 0.60 0.45 76% 0.47 2.0 0.004 500.0 1.20 0.93 77% 0.41 2.0 0.004 500.0 1.20 0.82 68% 0.50 2.0 0.004 500.0 1.20 i.oi 84% STANDARD DEVIATION: % RSD: STANDARD DEVIATION: % RSD: STANDARD DEVIATION: % RSD: STANDARD DEVIATION: % RSD: STANDARD DEVIATION: % RSD: STANDARD DEVIATION: % RSD: STANDARD DEVIATION: % RSD: 0.1787 11.8165 0.1072 8.1286 0.1072 9.4074 0.0666 7.1794 0.0975 10.3277 0.1942 22.3705 0.0795 10.3961 SERUM CURVE I (CARLTON) 7/27/95 y x 0.7212x + O@063 R 2. 0.967 1.20 owl ow 0,20 0.40 O.W O.W 100 1.20 140 MASS SPIKEG (,V) SERCRV2C.SUM Page I 0( )01()(-J' 152S-A.XLS 1995-08-22 13:41 OutPut of :950822B I 01)era(or DDW Date of theAnalysis 1995-08-22 09:04 Anal3sisFileName C:\SKALAR\DATA\IIWIDATA\SERUM\950822BI "n Inc I Tracer 1.50 1.46 98% 2 Drift 1.50 1.47 98% 3 Wash ND 4 Standard 1 0.015 0.015 103% 5 Standard 2 0.03 0.03 94% 6 Standard 3 0.06 0.06 105% 7 Standard 4 o@09 0.09 100% a Standard 5 0.12 0.12 98% 9 Standard 6 0.15 0.15 101% Standard 7 0.30 0.28 93% 11 Standard 8 0.60 0.62 103% 12 Standard 9 1.20 1.23 103% 13 Standard 10 l@50 1 @47 98% 14 Drift 1.50 1.48 99% Is Wash ND 16 SERUM BLK 1 0.07 17 SERUM BLK 2 0.04 18 SPK 62 -1 0.07 19 SPK 62-2 0.11 20 SPK 62-3 0.12 21 SPK 250-1 0.12 22 SPK 250-2 0.27 23 SPK 250-3 0.27 24 BLK 0.09 25 BLK 0.07 26 Drift 1.50 1.46 97% 27 Wash ND 28 F52713-1 0.07 29 F52718-29 0.06 30 F52723-1 0.05 2.0 0.10 2.0 0.10 2.0 0.10 2.0 0,10 2@O 0.10 2.0 0.10 2.0 0.10 2.0 0.10 2.0 0,10 2.0 0.10 2.0 0.10 2.0 0.10 2.0 0.10 Page 1 1.48 0,82 1,46 0.004 62.00 0.15 2.18 0.004 62.00 0.15 2.49 0.004 62.00 0.15 2.49 0.004 250.0 0.60 5.44 0.004 250.0 0.60 5.34 0.004 250.0 0.60 1.71 1.35 1.36 1.16 0.93 ................ #MEMIMmgm 31 F52715-1 0.05 32 F52738-1 0,06 33 F52809-1 0.07 34 F52724-1 0.06 35 F52737-1 0.06 36 F5280 1-1 0.05 37 F52729-1 0.04 38 Drift 1.50 1.46 98% 39 Wash ND 40 SPK 62-1 0.07 41 SPK 62-2 0.12 42 SPK 250-1 0.17 43 SPK 250-2 O@26 44 BLK 0.06 45 BLK C 0.04 46 SPK 62-1 0.08 47 SPK 62-2 0.08 48 SPK 250-1 0.20 49 SPK 250-2 0.22 50 Drift 1,50 1,48 99% 51 Wash ND 52 SPK 250-3 0.24 53 SPK 250-4 0.24 54 BLK 0.05 55 BLK 0.04 56 F52733-1 0,06 57 F52788-1 0.02 58 F52730-1 0.05 59 F5273 1-1 0.05 60 F52802-1 0.06 61 F52714-1 0.04 62 Drift 1.50 1.46 97% 63 Wash ND 64 F52734-1 0.05 65 F52774-1 0.06 66 F52712-1 0.06 152S-A.XLS #maw m 2.0 0.10 1.00 2.0 0.10 1.17 2.0 0.10 1.49 2.0 0.10 1.21 2.0 0.10 1.17 2.0 0.10 1.05 2,0 0.10 0.81 2.0 0.10 1.43 0.004 62.00 0.15 2.0 0,10 2.42 0.004 62.00 0.15 2.0 0.10 3.32 0.004 250.0 0.60 2.0 o@io 5.14 0.004 250@O O@60 2.0 0.10 1.16 2.0 0.10 0.88 2.0 0.10 1.59 0.004 62.00 0.15 2.0 0.10 1.66 0.004 62.00 0.15 2.p 0.10 4.06 0.004 250.0 0.60 2.0 0.10 4.38 0.004 250.0 0,60 2.0 0.10 4.86 0.004 250.0 0.60 2.0 0.10 4.76 0.004 250.0 0.60 2.0 0.10 1.03 2,0 0.10 0.72 2.0 0.10 1.11 2.0 0.10 0.32 2.0 0.10 1.04 2,0 0.10 0.95 2.0 0.10 1.11 2.0 0.10 0.87 2.0 0.10 1.07 2.0 0.10 1.23 2.0 0.10 1.28 Page 2 . ........ ........ 67 F52729-1 68 SPK 62-1 69 SPK 62-2 70 SPK 62-3 71 SPK 250-1 72 BLK 73 F52803-1 74 Drift 1.50 75 Wash 76 F52716-1 77 F52728-1 78 F52787-1 79 F52717-1 8() F52735-1 81 F52736-1 82 F5@726-1 83 F52732-1 84 F52739-1 85 SPK 62-1 86 Drift 1.50 87 Wash 88 SPK 62-2 89 SPK 250-1 9() SPK 250-2 91 Drift 1.50 92 Wash 0.()7 0.08 0.10 0.14 0.26 0.17 0.07 1.48 98% ND 0.07 0.07 0.06 0.09 0.07 0.06 0.05 O@07 0.05 0.08 1.50 100% ND 0.13 0.34 0.36 1.51 101% ND 152S-A.XLS 2.0 0.10 1.35 2.0 0.10 1.57 0.004 62.00 0.15 2.0 0,10 1.96 0.004 62.00 0.15 2.0 0.10 2.71 0.004 62,00 o@ 15 2.0 0.10 5.10 0.004 250.0 0.60 2.0 0.10 3.40 2.0 0.10 1.31 2.0 0.10 1.38 2.0 0.10 1.44 2.0 0.10 1.18 2.0 0.10 l@71 2.0 0.10 1.48 2.0 0.10 1.16 2.0 0.10 0.94 2.0 0.10 1.33 2.Q 0.10 0.90 2.0 0.10 1.68 0.004 62.00 0.15 2.0 0.10 2.66 0.004 62.00 0.15 2.0 0.10 6.72 0.004 250.0 0.60 2.0 0.10 7.28 0.004 250,0 0.60 Page 3 152S-B.XLS 1995-1)8-2311:10 OutPut of: 950822CI Operator :DDW Date ofthe Analysis: 1995-08-2213:41 Ana[3,siFsileName :C:\SKALAR\DATA\IIWIDATA\SERUM\950822CI I Traccr t.5() l@48 98% 2 Drift 1.50 1.49 99% 3 Wash ND 4 Standard 1 0.015 0.015 103% 5 Standard 2 0.03 0.03 94% 6 Standard 3 0.06 0.06 106% 7 Standard 4 o@09 0.09 98% 8 Standard 5 0.12 0.12 99% 9 Standard 6 o@ 15 0.15 101% 10 Standard 7 0.30 0.28 94% 11 Standard 8 0.60 0.61 102% 12 Standard 9 1,20 1.24 103% 13 Standard 10 1.50 1.46 97% 14 Drift 1.50 1.49 99% Is Wash ND 16 SERUM BLK 1 0.05 17 SERUM BLK 2 0.05 18 SPK 62-1 0.12 19 SPK 62-2 0.09 20 SPK 62-3 0.11 21 SPK 250-1 0.19 22 SPK 250-2 0.30 23 SPK 250-3 0.28 24 BLK 0,08 25 BLK 0.06 26 Drift 1.50 1.45 97% 27 Wasli ND 28 F5271 1-1 0.04 29 F52719-1 0.03 30 F52725-1 0.03 2.0 0.10 1.06 2.0 0.10 0.99 2.0 0.10 2@48 0,004 62 @00 0.15 2,0 0.10 1.77 0.004 62.00 0.15 2.0 0.10 2.24 0.004 62.00 0.15 2.0 0.10 3.74 0,004 250.0 0.60 2.0 o@ 10 5@98 0.004 250.0 0.60 2.0 0.10 5.50 0.004 250.0 0.60 2.0 0.10 1.65 2.0 0,10 1.11 2.0 0.10 0.84 2.0 0.10 0.68 2.0 0.10 0.66 Page I 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 5() 51 52 53 54 55 56 57 58 59 60 61 62 63 64 65 66 ............ F5272 1-1 0.03 F52727-1 0.02 F52806-1 0.03 F52711-2 0.02 F52719-2 0.02 F52725-2 0.02 F52721-2 ND Drift 1.50 1.49 99% Wash ND SPK 62-1 0.11 SPK 250-1 0.31 BLK 0.06 BLK 0.05 SPK 62-1 0.07 SPK 62-2 0.09 SPK 62-3 0.09 SPK 62-4 0.10 SPK 250-1 O@ 18 SPK 250-2 0.29 Drift 1.50 1.45 97% Wash ND SPK 250-3 0.27 BLK 0,06 BLK 0.04 F52727-2 O@03 F52806-2 0.03 F52713-2 0.02 F52718-2 0.02 F52723-2 0.03 F52715-2 0.03 F52738-2 0.02 Drift 1.50 1.53 102% Wash ND F52809-2 0.03 F52724-2 0.03 F52737-2 O@02 152S-B.XLS 2.0 0.10 0.51 2.0 0.10 0.48 2.0 0.10 0.53 2.0 0.10 0.42 2.0 0,10 0.34 2.0 0.10 0.31 2.0 o@ 10 ND 2.0 0.10 2.22 0.004 62.00 0.15 2.0 0.10 6,26 0.004 250.0 0.60 2.0 0.10 1.16 2,0 0.10 0.92 2.0 o@ 10 1.34 0.004 62.00 0.15 2.0 0.10 1.88 0.004 62.00 0.15 2.0 0.10 1.76 0.004 62.00 0.15 2.0 0.10 2.09 0.004 62.00 0.15 2.0 o@ 10 3.58 0.004 250.0 0.60 2.0 0.10 5.88 0.004 250.0 0.60 2.0 0.10 5.46 0.004 250.0 0.60 2.0 0.10 1.11 2.0 0.10 0.73 2.0 0.10 0.60 2.0 0.10 0.62 2.0 0.10 0.49 2.0 0.10 0.42 2.0 0.10 0.52 2.0 0.10 0.54 2.0 0.10 0.43 2.0 0.10 0.61 2.0 0.10 0.52 2.0 0,10 0,48 Page 2 .. .... 152S-B.XLS ........................... . ...... 67 SPK 62-1 0.05 2.0 0.10 1.03 0.004 62.00 0.15 68 SPK 62-2 0.07 2.0 0.10 1.39 0.004 62.00 0.15 69 SPK 250-1 0.14 2.0 0.10 2.88 0.004 250.0 0.60 70 SPK 250-2 0.24 2.0 0.10 4.70 0.004 250.0 0.60 71 Drift 1.50 1.51 101% 72 Wash ND Page 3 152S-C.XLS 1995-08-23 11:00 OutPut of: 950823AI Operator :DDW Date of theAnalysis: 1995-08-2306:55 Anal)-siFsileNanic C:\SKAI,AR\DATA\HWIDATA\SERUNB950823AI .... . ....... ..................... I Tracer 1.50 1.45 97% 2 Drift 1.50 1.47 98% 3 Wash ND 4 Standard 1 0.015 0.018 117% 5 Standard 2 0.03 0.03 89% 6 Standard 3 0.06 0.06 101% 7 Standard 4 0.09 0.09 102% 8 Standard 5 0.12 0.12 99% 9 Standard 6 0.15 1() Standard 7 O@30 0.15 1000/0 0.28 93% 11 Standard 8 0,60 0.62 103% 12 Standard 9 1.20 1.23 103% 13 Standard 10 1.5() 1.47 980/o 14 Drift 1.50 1.51 101% 15 Wash ND 16 SERUM BLK 0.06 2.0 17 SERUM BLK 0.05 2.0 18 SPK 62-1 0.08 2.0 19 SPK 62-2 0.10 2.0 20 SPK 62-3 0.10 2.0 21 SPK 250-1 0.14 2.0 22 SPK 250-2 0.21 2.0 23 SPK 250-3 O@23 2.0 24 SPK 250-4 0.23 2.0 25 SPK 250-5 0.26 2.0 26 Drift 1.50 1.51 100% 27 Wasli ND 28 SPK 250-6 0.39 2.0 29 BLK 0.07 2.0 30 BLK 0.03 2.0 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0,10 0,10 0.10 0.10 0.10 1.15 0.94 1.51 0.004 62.00 0.15 2.01 O@004 62.00 0.15 2.02 0.004 62.00 0.15 2.74 0.004 250.0 0.60 4.16 0.004 250.0 0.60 4.50 0.004 250.0 0.60 4.54 0.004 250.0 0.60 5.18 0.004 250,0 0.60 7.74 0.004 250.0 0.60 1.30 0.64 Page I 152S-C.XLS 31 F52801-2 0.05 2.0 0.10 0.92 32 F52720-2 0.04 2.0 0.10 0.73 33 F52733-2 0.03 2.0 0.10 0.64 34 F52788-2 0.04 2.0 0.10 0.74 35 F52730-2 0.03 2.0 0.10 0.59 36 1-52731-2 0.00 2.0 0.10 0.03 37 F52802-2 0.03 2.0 0.10 0.68 38 Drift 1.50 1.51 100% 39 Wash ND 40 F52714-2 0.04 2@O 0.10 0.85 41 F52734-2 0.03 2.0 0.10 0.69 42 F52774-2 0.04 2.0 0.10 0.72 43 SPK 62-1 o@io 2.0 0.10 1.92 0.004 62.00 0.15 44 SPK 250-1 45 SPK 250-2 0.15 0.17 2.0 0.10 2.92 0.004 250.0 0.60 2.0 0,10 3.46 0.004 250.0 0.60 46 BLK 0.05 2.0 0.10 1.03 47 BLK 0.05 2.0 0.10 1.03 48 SPK 62-1 0.05 2.0 0.10 1.03 0.004 62.00 0.15 49 SPK 62-2 0.08 2.0 0.10 1.64 0.004 62@00 0.15 50 Drift 1.50 1.53 102% 51 Wash ND 52 SPK 62-3 0.12 2.0 0.10 2.46 0.004 62,00 0,15 53 SPK 250-1 0.13 2.0 0.10 2.55 0.004 250.0 0.60 54 SPK 250-2 0.22 2.0 0.10 4.36 0.004 250.0 0,60 55 SPK 250-3 0.29 2.0 0.10 5.82 0.004 250.0 0.60 56 SPK 250-4 0.25 2.0 0.10 5.06 0.004 250.0 0.60 57 BLK 0.07 2.0 0.10 l@30 58 BLK 0.06 2.0 0.10 1.13 59 F52712-2 0.05 2.0 0.10 0.95 60 F52729-2 0.03 2.0 0.10 0.62 61 F52803-2 0.03 2.0 0.10 0.63 62 Drift 1.50 1.52 101% 63 Wash ND 64 F52716-2 0.03 2.0 o@io 0.65 ;A 65 F52728-2 0.04 2.0 (),10 0.77 66 F52787-2 0.03 2.0 0.10 0.68 Page 2 152S-C.XLS 67 F52717-2 0.03 2.0 0.10 0.62 68 SPK 62-1 0.05 2.0 0.10 0.97 0.004 62.00 0.15 69 SPK 62-2 0.09 2.0 0.10 1.83 0.004 62.00 0.15 70 SPK 250-1 o@ 10 2.0 0.10 2.06 0.004 250.0 0.60 71 SPK 250-2 0.24 2.0 0.10 4.76 0.004 250.0 0.60 72 SPK 250-3 0.18 2.0 0.10 3.64 0.004 250.0 0.60 73 Drift 1.5() 1.54 102% 74 Wash ND Page 3 152S-D.XLS 1995-08-23 15:22 OutPut of :950823BI Operator DDW Date of (heAnalysis: 1995-08-2310:50 AnalysisFileName :C:XSKALAR@DATA\IIWIDATA\SERUM\950823BI I Tracer l@50 1.48 98% 2 Drift 1.50 1.48 99% 3 Wash ND 4 Standard 1 0.015 0,016 106% 5 Standard 2 0.03 0.03 95% 6 Standard 3 0.06 0.06 101% 7 Standard 4 ().09 0.09 102% 8 Standard5 0.12 0.12 98% 9 Standard6 0.15 0.15 100% 1() Standard7 0.30 0.28 92% 11 Standard 8 0.60 0.62 104% 12 Standard9 1.20 1.23 102% 13 Standard 10 1.50 1.47 98% 14 Drift 1.50 1.49 99% 15 Wash ND 16 SERUM BLK 0.05 17 SERUM BLK 0.05 18 SERUM BLK 0.03 19 SPK 62-1 0.05 20 SPK 62-2 0.10 21 SPK 62-3 0.11 22 SPK 250-1 0.10 23 SPK 250-2 0.23 24 SPK 250-3 0.21 25 SPK 124-1 0.20 26 Drift l@50 1.47 98% 27 Wasli ND 28 SPK 124-2 0.21 29 BLK 0.13 30 BLK 0.06 2.0 0.10 1.04 2.0 0.10 0.93 2.0 0.10 0.67 2.0 0.10 1 @03 O@004 62.00 0.15 2.0 0.10 2.08 0.004 62.00 0.15 2.0 0.10 2.13 0.004 62.00 0.15 2.0 0.10 1,94 0.004 250.0 0.60 2.0 0.10 4.66 0.004 250.0 0.60 2@O 0.10 4.16 0.004 250.0 0.60 2.0 0.10 4,08 0,004 124.0 0.30 2.0 0.10 4.12 0.004 124.0 0.30 2.0 0.10 2.53 2,0 0.10 1.17 Page 1 152S-D.XLS ... old ............... Ik IVA 31 F52726-2 0.04 2.0 0.10 0.83 32 F52735-2 0.04 2.0 0.10 0.74 33 F52732-2 0.03 2.0 0.10 0.69 34 F52736-2 0.03 2.0 0.10 0.57 35 F52739-2 0,03 2.0 0.10 0.58 36 F52711-4 0.03 2.0 0.10 0.53 37 F52721-4 0.02 2.0 0.10 0.47 38 Drift 1.50 1.50 100% 39 Wash ND 40 F52719-4 0.26 2.0 0.10 5.10 41 F52727-4 0.04 2.0 0.10 0.70 42 F52725-4 0.03 2.0 0.10 0.60 43 F52723-4 44 SPK 124-1 45 BLK 46 BLK 47 SPK 62-1 48 SPK 62-2 49 SPK 62-3 0.06 0.17 0.04 0.04 0,06 0.08 0.10 2.0 0.10 1.25 2.0 0.10 3.30 0.004 124.0 0.30 2.0 0.10 0.89 2.0 0.10 0.84 2.0 0.10 1.14 0,004 62.00 0.15 2.0 0.10 1.67 0.004 62.00 0.15 2.0 0.10 2.06 O@004 62@00 0.15 50 Drift 1.50 1.48 99% 51 Wash ND 52 SPK 250-1 0.17 2.0 0.10 3.42 0.004 250.0 0.60 53 SPK 250-2 0.25 2@O o@io 4.96 0.004 250.0 0.60 54 SPK 250-3 0.29 2.0 0.10 5.88 0.004 250.0 0.60 55 BLK 56 BLK 0.11 2.0 0.10 2.23 0.08 2.0 0.10 1.57 57 BLK 0.02 2.0 0.10 0.48 58 F52726-2 0.04 2.0 0.10 0.75 59 F52732-2 ND 2.0 0.10 ND 60 F52739-2 ND 2.0 o@ 10 ND 61 F52735-2 0.02 2.0 0.10 0.43 62 Drift 1.50 1.45 96% %wo 63 Wash ND 64 F52736-2 0.03 2.0 0.10 0.57 65 F52711-4 ND 2.0 0.10 ND 66 BLK 0.11 2.0 0.10 2.25 Page 2 .......... 152S-D.XLS 67 BLK 0.18 2.0 0.10 3.64 68 BLK 0.04 2.0 0.10 0.79 69 BLK 0.02 2.0 0.10 0.42 70 SPK 62-1 0.08 2.0 0.10 1.58 0.004 62.00 0.15 71 SPK 62-2 0.08 2.0 0.10 1.65 0.004 62.00 0.15 72 SPK 250-1 0.17 2.0 0.10 3@48 0.004 250.0 0.60 73 SPK 250-2 0.18 2.0 0.10 3.56 0.004 250.0 0.60 74 Drift 1.50 1.49 100% 75 Wash ND 76 SPK 250-3 0.21 2.0 0.10 4.10 0.004 250.0 0.60 77 SPK 124-1 0.17 2.0 0.10 3.32 0,004 124.0 0.30 78 SPK 124-2 0.16 2.0 0.10 3.20 0.004 124.0 0.30 79 BLK 0.08 2.0 0,10 1.68 80 BLK 0.07 2.0 0.10 1.38 81 F52711-4 0.04 2.0 o@ 10 0.90 82 F527194 0.04 2.0 0.10 0.73 83 SPK 124-1 0.13 2.0 0.10 2.62 0.004 124.0 0.30 84 Drift 1.50 1.47 98% 85 Wash ND Lr, Page 3 152S-E.XLS 1995-08-28 15:03 OutPut of :951@828AI Operator :DDW Date of theAnalysis 1995-08-28 07:32 AnalysisFileNanic C:\SKALAR\DATA\IIWIDATA\SERUM\95082SAI I Traccr 1.50 1.45 97% 2 Drift l@50 1.47 98% 3 Wash ND 4 Standard 1 0.015 0.014 91% 5 Standard 2 0.03 0.03 105% 6 Standard 3 0.06 0.06 102% 7 Standard 4 0.09 0.09 99% 8 Standard 5 0.12 0.12 99% 9 Standard 6 0.15 0.15 100% 1() Standard 7 0.30 0.28 93% 11 Standard 8 0.60 0.62 103% 12 Standard 9 l@20 1.23 103% 13 Standard 10 1.50 1.47 98% 14 Drift 1.50 1.49 99% 15 Wash ND 16 SERUM BLANK O@06 17 SERUM BLANK 0.06 18 SPK 62-1 0.10 19 SPK 62-2 0.09 20 SPK 250-1 0.20 21 SPK 250-2 0.21 22 SPK 124-1 0.19 23 SPK 124-2 0.18 24 BLANK 0.06 25 F527254 0.07 26 Drift l@50 1.46 97% 27 Wasli ND 28 F52721-4 0.06 29 F52727-4 0.05 30 F52806-4 0.05 2.0 0.10 1.15 2.0 0,10 1.15 2.0 0.10 1.92 0.004 62.00 0.15 2.0 0.10 l@73 0,004 62.00 0.15 2.0 0.10 3.90 0.004 250.0 0,60 2.0 0.10 4.28 0.004 250.0 0.60 2.0 0.10 3.84 0.004 124.0 0.30 2.0 0.10 3.52 0.004 124.0 0.30 2.0 0.10 1.20 2.0 0.10 1.32 2.0 0.10 1.21 2.0 0.10 0.93 2.o 0.10 0,92 Page 1 152S-E.XLS .......................... ........... 31 F52713-4 0.03 2.0 0.10 0.64 32 F52718-4 0.05 2.0 0.10 0.99 33 F52723-4 0.04 2.0 0.10 0.73 34 F52715-4 0.03 2.0 0.10 0.69 35 F52738-4 0.07 2.0 0.10 1.40 36 SPK 62-1 0.05 2@O o@io 0.93 0,004 62.00 0.15 37 SPK 62-2 0.12 2.0 0.10 2.30 0.004 62.00 0.15 38 Drift 1.50 1.48 99% 39 Wash ND 40 SPK 124-1 0.15 41 BLANK-1 0.05 2.0 0.10 2.93 0.004 124.0 0.30 2.0 0.10 1.04 42 BLANK-2 0.04 43 SPK 62-1 0.08 44 SPK 62-2' 0.08 45 SPK 250-1 0.30 2.0 0.10 0.75 2@O 0.10 l@58 0.004 62.00 0.t5 2.0 0.10 1.69 0.004 62.00 o@ 15 2.0 0.10 6.04 0.004 250.0 0.60 46 SPK 250-2 0.26 2.0 0.10 5.14 0.004 250.0 0.60 47 BLK 0.06 2.0 0.10 1,21 48 F52809-4 0.04 2.0 0.10 0.72 49 F52724-4 0.03 2.0 0.10 0.64 50 Drift 1.50 1.47 98% 51 Wash ND 52 F52737-4 0,03 2.0 0.10 0.64 53 F52801-4 0.06 2.0 0.10 1.20 54 F52720-4 0.05 2.0 o@ 10 0.92 55 F52733-4 0.04 2.0 0.10 0.70 56 F52788-4 0.02 2.0 0.10 0.50 57 F52730-4 0.02 2.0 0.10 0.39 58 F52731-4 0.02 2.0 0.10 0.39 59 F52802-4 0.03 2.0 0.10 0.59 60 SPK 62-1 0.05 2.0 0.10 0.95 0.004 62.00 0.15 61 SPK 62-2 o@io 2.0 0.10 2.05 0.004 62.00 0.15 62 Drift 1.50 1.49 99% 63 Wash ND 64 SPK 250-1 0.18 65 SPK 250-2 0.24 2.0 o@ 10 3.52 0.004 250.0 0.60 2.0 o@ 1() 4,70 0.004 250.0 0.60 66 SPK 124-1 0.16 2.0 0.10 3.20 0.004 124.0 0.30 Page 2 152S-E.XLS 67 68 69 70 71 72 73 74 75 76 77 78 79 8() 81 82 83 84 85 BLK BLK F52714-4 F52734-4 F52774-4 F52712-4 F52729-4 Drift Wash F52803-4 F52716-4 F52728-4 F52787-4 SPK 62-1 SP K 62-2 SPK' 124-1 SPK 124-2 Drift Wash 0.08 0.06 0.05 0.07 0.05 0.03 O@03 1.50 1.49 99% ND 0.05 0.04 0.03 0.04 0.07 0.11 0.09 0.15 1.50 1.51 100% ND 2.0 0.10 1.62 2.0 0.10 1.25 2.0 0.10 0.93 2.0 0.10 1.35 2.0 0,10 0.92 2.0 0,10 0.69 2.0 0.10 0.67 2.0 0.10 0.92 2.0 0.10 0.77 2.0 0.10 0.68 2.0 0.10 0.76 2.0 0.10 1.31 0.004 62.00 0,15 2.0 0.10 2@20 0.004 62.00 0.15 2.0 0.10 1.81 0.004 124.0 O@30 2.0 0.10 2.97 0.004 124.0 0.30 Page 3 152S-F.XLS 1995-1)8-2906:38 OutPut of: 950828B I Operator DDW Date ofthe Analysis:1995-08-28 11:47 Anal)-siFsileNante C:\SKALAR\DATA\IIWIDATA\SERUM\950828BI I Tracer l@50 l@46 98% 2 Drift 1.50 1.47 98% 3 Wash ND 4 Standard 1 0.015 0.011 74% 5 Standard 2 0.03 0.04 131% 6 Standard 3 0.06 0.06 97% 7 Standard 4 0.09 0.09 96% 8 Standard 5 0.12 0.12 99% 9 Standard 6 0.15 0.15 101% Standard 7 0.30 0.28 93% 11 Standard 8 0.60 0.62 103% 12 Standard 9 1.20 1.23 102% 13 Standard 10 1,50 1.47 98% 14 Drift 1.50 1.50 100% Is Wash ND 16 SERUM BLK 1 0.04 17 SERUM BLK 2 0.03 18 SPK 62-1 19 SPK 62-2 0.11 0.09 20 SPK 124-1 0.29 21 SPK 124-2 0.13 22 BLK 0.06 23 BLK 0.04 24 F52717-4 0.05 25 F52735-4 0.03 26 Drift 1.50 1.46 97% 27 Wash ND 28 F52736-4 0.03 29 F527264 0.03 30 F52732-4 0.02 2.0 0.10 0.84 2.0 0.10 0.69 2.0 0.10 2.16 0.004 62.00 0.15 2.0 0.10 1.89 0.004 62.00 0.15 2.0 0.10 5.86 0,004 124.0 0.30 2.0 0.10 2.55 0.004 124.0 0.30 2.0 0.10 1.20 2.0 0.10 0.71 2.0 0.10 1.03 2.0 0.10 0.67 2.0 0,10 0.64 2.0 0.10 0.54 2@O 0.10 0.42 Page I 152S-F.XLS ........m.i... A.WW 31 F52739-4 0.02 2.0 0.10 0.33 32 F52711-8 0.02 2@O 0.10 0.45 33 F52719-8 ND 2.0 0,10 ND 34 BLK 0.03 2.0 0.10 0.60 35 SPK 62-1 0.05 2.0 0.10 1.09 0.004 62.00 0,15 36 SPK 62-2 0.06 2.0 o@io 1.17 0.004 62.00 0.15 37 SPK 124-1 0.08 2.0 0.10 1.58 0.004 124.0 0.30 38 Drift 1.50 1.46 97% 39 Wash ND 40 SPK 124-2 0.17 41 BLK o@ll 42 BLK 0.05 43 F52725-8 0.04 2,0 0.10 3.48 0.004 124,0 0.30 2.0 0.10 2.15 2.0 0.10 0.93 2.0 0.10 O@78 44 F5272 1-8 0.03 2.0 0.10 0.63 45 F52727- 0.03 2.0 0.10 0.53 46 F52806-8 O@02 2.0 0.10 0.49 47 F52713-8 0,02 2.0 0.10 0.31 48 F52718-8 0.02 2.0 0.10 0.33 49 F52723-8 0.02 2.0 0.10 0.32 50 Drift 1.50 1.46 98% 51 Wash ND 52 F52715-8 O@02 2.0 0.10 0,49 53 F52738-8 0.02 2.0 0.10 0.38 54 F52809-8 55 SPK 62-1 56 SPK 62-2 57 SPK 124-1 58 SPK 124-2 59 BLK 0.02 0.05 0.06 0.10 0.12 0.04 2.0 0.10 0.33 2.0 0.10 0.90 0.004 62.00 0.15 2.0 0.10 1.30 0.004 62.00 0.15 2.0 0.to 1.98 0.004 124.0 0.30 2.0 0.10 2.44 0.004 124.0 0.30 2.0 0,10 0.72 60 BLK 0.02 2.0 0.10 0.39 61 SPK 62-1 O@08 2.0 0.10 1.50 0.004 62.00 0.15 62 Drift 1.50 1.46 97% 63 Wash ND 64 SPK 62-2 0.08 65 SPK 124-1 0.14 66 SPK 124-2 0.14 2.0 0.10 1.60 0.004 62.00 0.15 2.0 0.10 2@79 0,004 124.0 0.30 2.0 0.10 2.85 0.004 124.0 0.30 Page 2 152S-F.XLS .. ....... 67 BLK 0.05 2.0 0.10 0.95 68 BLK 0.04 2.0 0.10 0.76 69 F52724-8 0.04 2.0 0.10 0.74 70 F52737-8 0.04 2.0 0.10 0.72 71 F52801-9 0.06 2.0 0,10 1.29 72 F52720-8 0.05 2.0 0.10 0.92 73 F52733-8 0.03 2,0 0.10 0.54 74 Drift 1.50 1.47 98% 75 Wash ND 76 F52788-8 ND 2.0 0.10 ND 77 SPK 62-1 0.06 2.0 0.10 1.12 0.004 62.00 0.15 78 SPK 62-2 0.07 2.0 0.10 1.31 0.004 62.00 0.15 79 SPK 124-1 O.It 2.0 0.10 2.15 0,004 124.0 0.30 80 Drift 1.5() 1.47 98% 81 Wash ND Page 3 1995-Oik.---22,3:41 Software : version 6.1 OutPut of : 950822Bl cl990,93 Operator : DDW Date of the Analysis : 1995-08-22 09:04 Analysis File Name : C:\SKALAR\DATA\HWIDATA\SERUM\950822Bl Fluoride 1.5 Calibration order = Inverse Logarithm Slope : 8 = #.I#### lo[ x - cl Result 8 a2 = al = aO = -0.00000 0.00072 -1.20078 x = corrected value of the sample cl = corrected value of the concentration 1 s = Slope of the electrode Fluoride L Calibration order = 2 Coz,.relation r = 0.99917 Result = a2 x2 + al * x + aO a2 -0.00000 al 0.00025 aO 0.00551 Sampler Diluter Type Number Sample Time Wash Time Air Time Take up special needle Height : SA1000 :1 : 50 sec. : 120 sec. : 1 Sec. : Single : None : 70 mm. needle Height : 80 mm dilution Factor : 10 dilution Volume : 2.5 ml. Resample :1 Dilution runs : 1 User file : TXT Reproces : No 000125 1995-08.-22 13:41 OutPUt Of Fluoride 1.5 Path number Signal type Decolor 3 : Debubbled : yes system Number o dilute No Resample No dil Threshold 4095 diG output 0 Window event : off al standard : s2 standard : s3 standard : s4 standard : s5 standard : s6 standard : s7 standard : s8 standard : 89 standard : slO standard : Order : Inverse Dimension : Ppm Ignore Ignore Ignore Ignore Ignore 0.150 0.300 0.600 1.200 1.500 Logarithm start Value trigger Limit Peak shape StArt ignore eNd ignore Measure window Filter Regeneration formula : 500 DU : 1800 Sgc : Pointed : -.60 Sec : 120 Sec : 75 % : No : No output Fluoride L Path number Signal type Decolor system Number dilute Resample dil Threshold diG output Window event 0 Debubbled No 0 No No 4095 0 Off 950822BI 0001IZ6 1995-08@-22 13:41 OutPut of : 950822Bl sl standard : s2 standard : s3 standard : 0.015 0.030 0.060 s4 standard : 0.090 s5 standard : 0.120 s6 standard : 0.150 s7 standard : Ignore ss standard : Ignore 89 standard : Ignore sio standard : Ignore Order : 2 Dimension : PPM start Value : 500 DU trigger Limit : 1800 Sec Peak shape : Pointed start ignore : 60 Sec eNd ignore : 120 Soc measure window : 75 t Filter : No Regeneration : No formula c4:=c3 output ooolz7 1995-OS.-22 13:41 OutPut Of : 950822131 Fluoride 1.5 Fluoride L PPM ppm Pos Typ Ident Ch Result F Time Ch Result F Time wt iw 1t 2d 3w 4 sl 5 s2 6 s3 7 s4 8 s5 9 s6 10 s7 11 s8 12 s9 13 slO 14 d 15 w 16 u 17 u 18 u 19 u 20 u 21 u 22 u 23 u 24 u 25 u 26 d 27 w 28 u 29 u 30 u 31 u 32 u 33 u 34 u 35 u 36 u 37 u 38 d 39 w 40 u 41 u 42 u 43 u 44 u 45 u 46 u 47. u 48 u 49 u 50 - d 51 w 52 u 53 u Initial Wash 3 0.063 Tracer 3 1.464 Drift 3 1.468 Wash 3 0.063 Standard 1 3 0.067 Standard 2 3 0.073 Standard 3 3 0.091 Standard 4 3 0.108 Standard 5 3 0.127 Standard 6 3 0.157 Standard 7 3 0.278 Standard 8 3 0.618 Standard 9 3 1.232 Standard 10 3 1.466 Drift 3 1.480 Wash 3 0.063 SERUM BLK 1 3 0.098 SERUM BLK 2 3 0.079 SPK 62-1 3 0.097 SPK 62-2 3 0.121 SPK 62-3 3 0.133 SPK 250-1 3 0.133 SPK 250-2 3 0.272 SPK 250-3 3 0.267 BLK 3 0.105 BLK 3 0.094 Drift 3 1.457 Wash 3 0.063 F52713-1 F52718-29 P52723-1 F52715-1 F52738-1 3 0.094 3 0.088 3 0.082 3 0.084 3 0.089 F52809-1 3 0.098 F52724-1 3 O.Ogo F52737-1 P52801-1 P52729-1 3 0.089 3 0.085 3 0.079 Drift 3 Wash 3 SPK 62-1 3 SPK 62-2 3 SPK 250-1 3 SPK.250-2 3 1.464 0.063 0.096 0.131 0.1660.257 BLK 3 0.088 BLK C 3 0.081 SPX 62-1 3 0.101 SPK 62-2 3 0.103 SPK 250-1 3 0.203 SPK 250-2 3 0.219 Drift 3 1.479 Wash 3 0.063 SPK 250-3 3 0.243 SPK 250-4 3 0.238 65 4 0.0055 210 4 0.6754 384 4 0.6770 626 4 0.0055 731 4 0.0154 909 4 0.0282 1085 4 0.0629 1257 4 0.0902 1435 4 0.1171 1611 4 0.1512 1786 4 0.2529 1960 2136 2310 2486 4 0.4152 4 0.6046 4 0.6763 4 0.6810 2727 2837 4 0.0055 4 0.0738 3007 4 0.0410 3185 4 0.0730 - 3363 4 0.1088 3539 4 0.1246 3713 4 0.1246 3888 4 0.2488 4063 4237 4 0.2453 4 0.0856 4416 , 4 0.0673 4588 4 0.6732 4823 4 0.0055 4938 4 0.0681 5113 4 0.0582 5289 5461 5635 4 0.0467 4 0.0502 4 0.0584 5813 4 0.0743 5991 4 0.0604 6163 4 0.0587 6340 4 0.0527 6515 4 0.0405 6690 4 0.6754 6929 4 0.0055 7042 7229 4 0.0715 4 0.1212 7392 7566 7741 4 0.1612 4 0.2380 4 0.0579 7917 4 0.0440 8092 8267 8441 8617 8792 9031 9142 9318 4 0.0794 4 0.0829 4 0.1957 4 0.2097 4 0.6807 4 0.0055 4 0.2278 4 0.2241 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Page 1 of 2 oooilzs 1995-08:--22 13:41 OutPut of : 950822BI Fluoride 1.5 Fluoride L ppm ppm Pos Typ Ident Ch Result P Time Ch Result F Time 54 u BLK 3 0.085 9490 4 0.0517 0 55 u ELK 3 0.077 9668 4 0.0362 0 56 u P52733-1 3 0.087 9842 4 0.0554 0 57 u F52788-1 3 0.068 10017 4 0.0161 0 58 u P52730-1 3 0.085 10193 4 0.0520 0 59 u F52731-1 3 0.083 10367 4 0.0477 0 60 u P52802-1 3 0.087 10545 4 0.0557 0 61 u F52714-1 3 0.081 10719 4 0.0435 0 62 d Drift 3 1.462 10893 4 0.6748 0 63 w Wash 3 0.063 11126 4 0.0055 0 64 u P52734-1 3 0.086 11240 4 0.0537 0 65 u F52774-1 3 0.090 11417 4 0.0617 0 66 u F52712-1 3 0.092 11596 4 0.0641 0 67 u P52729-1 3 0.094 11785 4 0.0676 0 68 u SPK 62-1 3 0.100 11945 4 0.0785 0 69 u SPR 62-2 3 0.114 12120 4 0.0981 0 70 u SPK 62-3 3 0.143 12294 4 0.1356 0 71 u SPK 250-1 3 0.255 12469 4 0.2371 0 72 u ELK 3 0.170 12645 4 0.1650 0 73 u F52803-1 3 0.093 -.12820 4 0.0654 0 74 d Drift 3 1.475 12995 4 0.6793 0 75 -w Wash 3 0.063 13230 4 0.0055 0 76 u F52716-1 3 0.095 13345 4 0.0691 0 77 u F52728-1 3 0.097 13521 4 0.0720 0 78 u P52787-1 3 0.089 13695 4 0.0592 0 79 u F52717-1 3 0.105 13871, 4 0.0853 0 80 u F52735-1 3 0.098 14045 4 0.0738 0 81 u F52736-1 3 0.088 14221 4 0.0579 0 82 u F52726-1 3 0.083 14396 4 0.0472 0 83 u F52732-1 3 0.093 14571 4 0.0666 0 84 u F52739-1 3 0.081 14745 4 0.0450 0 85 u SPK 62-1 3 0.104 14920 4 0.0839 0 86 d Drift 3 1.503 15097 4 0.6888 0 87 w Wash 3 0.063 15338 4 0.0055 0 88 u SPK 62-2 3 0.140 15450 4 0.1328 0 89 u SPK 250-1 3 0.336 15621 4 0.2886 0 90 u SPK 250-2 3 0.364 15800 4 0.3039 0 91 d Drift 3 1.508 15972 4 0.6906 0 92 w Wash 3 0.063 16213 4 0.0055 0 wt rw RunOut Wash 3 0.063 16447 4 0.0055 0 Page 2 of 2 0.2236693 Calibration curve of 95082281 Fluoride L 1+ @0-0055094 0 Order 2 Measured r 0.99917 900 1 000130 1.7087549- Calibration curve of 95082281 Fluoride 1.5 06361170 IM-IF.N64 M c 0. 0629813 0 Order Measured Inverse Logarithm 409S s 6378 oooizi:t Raw data of 950822BI : Fluoride I.S ttG 409S 0 Time 0 Esc=Exit 1 FI=Help 1 Crtl-P=Edit peaks 1 'til I I Z*i 5000 00013L@, Raw data of 950822BI : Fluoride I.S ELT-IIMMEEMNUE3L4560 E"-TM-30241o Fi?-JTM-6 4095 1 2 0 4560 Esc=Exit : FI=Help Time Crtl-P=Edit peaks Iz 9560 0()0133 Raw data of 9SO822BI : Fluoride I.S NNW -M 240 IF$?-3YM 94 409S Li P4 '7 0 to 4 \Aj 0 OS3S Time Esc=Exit : FI=Help 1 Crtl-P=Edit peaks 1393S 000134 Raw data of 9SO822BI : Fluoride I.S i331OWnll-M 272 F#7-31PU-s 4095 JU o@\ 90 0 13310 Time Esc=Exit 1 FI=Help 1 Crtl-P=Edit peaks i 18310 000135 1995-@08-22 17;22 Sof'tware version 6.1 OutPut of cl990,93 950822Cl Operator : DDW Date of the Analysis : 1995-08-22 13:41 Analysis File Name : C:\SKALAR\DATA\HWIDATA\SBRUM\950822Cl t0 elS OLJ-= (.o3Z9 -IS &r V --% (-AA.L404 cai-@W- Fluoride 1.5 Calibration order = Inverse Logarithm Slope Result lo[ x cl s a2 = al = ao = -0.00000 0.00071 -1-19998 x = corrected value of the sample cl = corrected value of the concentration 1 s = Slope of the electrode Fluoride L Calibration order 2 Coz,rolation r = o.99_910 Result a2 al aO = a2 Xa + al * x + aO 0.00000 0.00023 0.00817 Sampler Diluter Type Number Sample Time Wash Time Air Time Take up special needle Height SA1000 1 50 sec. 120 sec. 1 sec. Single None 70 mm. needle Height : 80 mm dilution Factor : 10 dilution volume : 2.5 ml. Resample :1 Dilution runs User file : TXT Reproces : No 000136 1995-08-22 17:22 OutPut of Fluoride 1.5 Path number 3 Signal type : Debubbled Decolor : yes system Number : 0 diluts : No Resample : No -dil Threshold : 4095 diG output :0 Window event : Off ol standard : s2 standard : s3 standard : s4 standard : 85 standard : s6 standard : s7 standard : S8 standard : 89 standard : slO standard : Order : Inverse Dimension : PPM start Value trigger Limit Peak shape start ignore eNd ignore Measure window Filter Regeneration formula output Ignore Ignore Ignore Ignore Ignore 0.150 0.300 0.600 1.200 1.500 Logarithm 500 DU 1800 Soc Pointed 60 Sec 120 Sec 75 t No No Fluoride L Path number Signal type Decolor system Number diluto Resample dil Threshold diG output Window event 0 : Debubbled : No :0 : No : No : 4095 :0 : Off 950822Cl 1995-@-08-22 17:22 OutPut of el standard : 0.015 s2 standard : 0.030 o3 standard : 0.060 s4 standard : 0.090 85 standard : s6 standard : 0.120 0.150 s7 standard : Ignore s8 standard : Ignore S9 standard : Ignore slO standard : Ignore Order : 2 Dimension : PPM start Value : 500 DU trigger Limit : 1800 Sec Peak shape start ignore eNd ignore : Pointed : 60 Sec : 120 Sec Measure window : 75 t Filter : No Regeneration : No formula c4:=c3 output 950822CI 0 (10 1 a 1995-08-22 17:22 OutPut of : 950822Cl Fluoride 1.5 Fluoride L PPM ppm Pos Typ Ident Ch Result F Time Ch Result F Time wt iw Initial Wash 3 0.063 65 4 0.0082 0 1t Tracer 3 1.476 210 4 0.9301 0 2d Drift 3 1.488 386 4 0.9386 0 3w Wash 3 0.063 619 4 0.0082 0 4 al Standard 1 3 0.066 734 4 0.0154 0 5 s2 Standard 2 3 0.073 911 4 0.0282 0 6 s3 Standard 3 3 0.092 1086 4 0.0636 0 7 s4 Standard 4 3 0.108 1261 4 0.0885 0 8 s5 Standard 5 3 0.129 1435 4 0.1183 0 9 s6 Standard 6 3 0.156 1611 4 0.1510 0 10 s7 Standard 7 3 0.281 1785 4 0.2692 0 11 s8 Standard 8 3 0.614 1959 4 0.4788 0 12 s9 Standard 9 3 1.238 2134 4 0.7912 0 13 slO Standard 10 3 1.462 2308 4 0.9213 0 14 d Drift 3 1.488 2483 4 0.9386 0 15 w Wash 3 0.063 2723 4 0.0082 0 16 u SERUM BLK 1 3 0.086 2831 4 0.0528 0 17 u SERUM BLK 2 3 0.084 3006 4 0.0493 0 18 u SPK 62-1 3 0.134 3184 4 0.1240 0 19 u SPK 62-2 3 0.108 3360 4 0.0885 0 20 u SPK 62-3 3 0.124 3533 4 0.1120 0 21 u SPK 250-1 3 0.187 3710 4 0.1854 0 22 u SPK 250-2 3 0.299 3884 4 0.2835 0 23 u SPK 250-3 3 0.275 4060 4 0.2644 0 24 u BLK 3 0.104 4231 4 0.0823 0 25 u BLK 3 0.087 4408 4 0.0556 0 26 d Drift 3 1.449 4584 ' 4 0.9125 0 27 w Wash 3 0.063 4789 4 0.0082 0 28 u F52711-1 3 0.080 4935 4 0.0418 0 29 u F52719-1 3 0.076 5111 4 0.0340 0 30 u F52725-1 3 0.075 5283 4 0.0328 0 31 u F52721-1 3 0.071 5459 4 0.0257 0 32 u F52727-1 3 0.071 5633 4 0.0239 0 33 u F52806-1 3 0.072 5811 4 0.0264 0 34 u F52711-2 3 0.069 5977 4 0.0209 0 35 u F52719-2 3 0.067 6155 4 0.0172 0 36 u F52725-2 3 0.066 6335 4 0.0154 0 37 u F52721-2 3 0.063 6509 4 0.0088 0 38 d Drift 3 1.489 6685 4 0.9390 0 39 w Wash 3 0.063 6926 4 0.0082 0 40 u SPK 62-1 3 0.124 7033 4 0.1112 0 41 u SPK 250-1 3 0.313 7213 4 0.2938 0 42 u BLK 3 0.089. 7384 4 0.0581 0 43 u BLK 3 0.082 7558 4 0.0460 0 44 u SPK 62-1 3 0.094 7736 4 0.0671 0 45 u SPK 62-2 3 0.112 -7911 4 0.0940 0 46 u SPK 62-3 3 0.108 8084 4 0.0880 0 47 u SPK 62-4 3 0.119 8261 4 0.1047 0 48 u SPK 250-1 3 0.179 8434 4 0.1767 0 49 u SPK 250-2 3 0.294 8611 4 0.2791 0 50 d Drift 3 1.454 8785 4 0.9157 0 51 w Wash 3 0.063 9025 4 0.0082 0 52 u SPK 250-3 3 0.273 9136 4 0.2630 0 53 u BLK 3 0.087 9309 4 0.0556 0 Page 1 of 2 OOOJL39 1995--:08-22 17:22 OutPut of : 950822Cl Fluoride 1.5 Fluoride L ppm PPM Pos Typ Ident Ch Result F Time Ch Result F Time 54 u BLK 3 0.077 9482 4 0.0367 0 55 u F52727-2 3 0.074 9658 4 0.0300 0 56 u F52806-2 3 0.074 9838 4 0.0312 0 57 u F52713-2 3 0.071 10004 4 0.0245 0 58 u F52718-2 3 0.069 10184 4 0.0209 0 59 u F52723-2 3 0.072 10351 4 0.0261 0 60 u F52715-2 3 0.072 10536 4 0.0271 0 61 u F52738-2 3 0.069 10711 4 0.0216 0 62 d Drift 3 1.526 10886 4 0.9653 0 63 w Wash 3 0.063 11127 4 0.0082 0 64 u F52809-2 3 0.074 11234 4 0.0305 0 65 u F52724-2 3 0.072 11413 4 0.0261 0 66 u F52737-2 3 0.071 11587 4 0.0241 0 67 u SPK 62-1 3 0.085 11761 4 0.0514 0 68 u SPK 62-2 3 0.096 11936 4 0.0697 0 69 u SPK 250-1 3 0.150 12112 4 0.1442 0 70 u SPK 250-2 3 0.235 12286 4 0.2310 0 71 d Drift 3 1.512 12462 4 0.9553 0 72 w Wash 3 0.063 12703 4 0.0082 0 wt rw RunOut Wash 3 0.063 .12937 4 0.0082 0 Page 2 of 2 000140 0.2278068@ Calibration curve of 950822CI Fluoride L c .008 16!530 Order 2 Measured r 0.99910 900 00@ 000141 lot 1.7318146 Calibration curve of 950822CI 6 Fluoride 1.5 0.0637 c 0.0630991 0 Order Measlured 1 Inverse Logarithm 4095 s 6271 000142 qo Raw data of 950822CI : Fluoride 1.5 0 136 64 4095 0 0 Esc=Exit FI=Help T ime Crtl-P=Edit peaks 1 500*0 oool43 1995-08-23 11:00 OutPut of : 950823Al Software : version 6.1 cl990,93 Operator : DDW Date of the Analysis : 1995-08-23 06:55 Analysis File Name : C:\.SKALAR\DATA\KWIDATA\SERUM\950823Al 4m b,-r CA'3 7.9 M -c- Fluoride 1.5 Calibration order = Inverse Logarithm Slope 8 = #.##### x Result = 101 a2 = al = aO = -0.00000 0.00076 -1.19531 cl 1 x = corrected value of the sample cl = corrected value of the concentration 1 s = Slope of the electrode Fluoride L Calibration order = 2 Correlation r = 0.99897 Result = a2 X2 a2 -0.00000 al 0.00028 aO 0.00600 + al * x + aO Sampler Diluter Type Number Sample Time Wash Time Air Time Take up special needle Height : SA1000 :1 : 50 sec. : 120 sec. : 1 sec. : Single : None : 70 mm. needle Height : 80 mm dilution Factor : 10 dilution Volume : 2.5 ml. Resample :I Dilution runs : 1 User file : TXT Reproces : No 000144 1995-08-23 11:00 output of Fluoride 1.5 Path number Signal type Decolor system Number diluto Resample dil Threshold diG output Window event 3 : Debubbled : Yes :0 : No : No : 4095 :0 : Off al standard : s2 standard : s3 standard : s4 standard : s5 standard : s6 standard : s7 standard : s8 standard : 89 standard : slO standard : Order : Inverse Dimension : PPM start Value trigger Limit Peak shape start ignore eNd ignore Measure window Filter Regeneration formula output Ignore Ignore Ignore Ignore Ignore 0.150 0.300 0.600 1.200 1.500 Logarithm 500 DU 1800 Sec Pointed 60 Soc 120' Sec 75 0 No No Fluoride L Path number Signal type Decolor system Number diluto Rezample dil Threshold diG output Window event :0 : Debubbled : No :0 : No : No : 4095 :0 : Off 950823Al oool45 43 1995-08-23 11:00 OutPut of sl standard : s2 standard : 0.015 0.030 s3 standard : 0.060 s4 standard : 0.090 85 standard : 0.120 s6 standard : 0.150 s7 standard : Ignore S8 standard : Ignore 89 standard : Ignore 810 standard : Ignore Order : 2 Dimension : PPM start Value : 500 DU trigger Limit : 1800 sac Peak shape start ignore eNd ignore : Pointed : 60 Sec : 120 Sec Measure window : 75 Filter : No Regeneration : No formula C4:=c3 output 950823Al 000146 qq 1995@-08-23 11:00 OutPut of : 950823Al Page 1 of 2 Fluoride 1.5 Fluoride L ppm PPM Pos Typ Ident Ch Result F Time Ch Result F Time wt iw 1t 2d 3w 4 el 5 s2 6 s3 7 s4 8 S5 9 s6 10 s7 11 S8 12 s9 13 siO 14 d 15 w 16 u 17 u 18 u 19 u 20 u 21- u 22 u 23 u 24- u 25 u 26 d 27 w 28 u 29 u 30 u 31 u 32 u 33 u 34 u 35 u 36 u 37 u 38 d 39 w 40 u 41 u 42 u 43 u 44 u 45 u 46 u 47 u 48 u 49 u 50 d 51 w 52 u 53 u Initial Wash 3 Tracer 3 Drift 3 Wash 3 Standard 1 3 Standard 2 3 Standard 3 3 Standard 4 3 Standard 5 3 Standard 6 3 Standard 7 3 Standard 8 3 Standard 9 3 Standard 10 3 Drift 3 Wash 3 SERUM BLK 3 SERUM BLK 3 SPK 62-1 3 SPK 62-2 3 SPK 62-3 3 SPK 250-1 3 SPK 250-2 3 SPK 250-3 3 SPK 250-4 3 SPK 250-5 3 Drift 3 Wash 3 SPK 250-6 3 BLK 3 BLK 3 P52801-2 3 F52720-2 3 F52733-2 3 F52788-2 3 F52730-2 3 F52731-2 3 F52802-2 3 Drift 3 Wash 3 P52714-2 3 F52734-2 3 P52774-2 3 SPK 62-1 3 SPK 250-1 3 SPK 250-2 3 BLK 3 BLK 3 SPK 62-1 3 SPK 62-2 3 Drift 3 Wash 3 SPK 62-3 3 SPK 250-1 3 0.064 1.452 1.469 0.064 0.069 0.073 0.090 0.109 0.129 0.157 0.278 0.618 1.233 1.466 1.510 0.064 0.088 0.082 0.098 0.115 0.115 0.144 0.208 0.225 0.227 0.259 1.505 0.064 0.387 0.092 0.075 0.082 0.077 0.075 0.077 0.074 0.062 0.076 1.506 0.064 0.080 0.076 0.0770.112 0.153 0.173 0.085 0.085 0.085 0.102 1.529 0.064 0.132 0.136 65 209 384 615 732 910 1086 1260 1436 1610 1786 1960 2134 2310 2485 2724 2833 3010 3186 3361 3533 3711 3887 4063 4237 4412 4587 4819 4937 5112 5284 5462 5636 5812 5986 6162 6338 6513 6688 6930 7037 7215 7391 7565 7740 '7915 8089 8264 8438 8616 8790 9025 9140 9314 4 0.0060 4 0.5049 4 0.5067 4 0.0060 4 0.0175 4 0.0266 4 0.0605 4 0.0914 4 0.1186 4 0.1503 4 0.2419 4 0.3698 4 0.4794 4 0.5064 4 0.5109 4 0.0060 4 0.0576 4 0.0468 4 0.0753 4 0.1004 4 0.1012 4 0.1371 4 0.1954 4 0.2081 4 0.2094 4 0.2308 4 0.5104 4 0.0060 4 0.2952 4 0.0651 4 0.0321 4 0.0458 4 0.0365 4 0.0321 4 0.0371 4 0.0294 4 0.0015 4 0.0341 4 0.5106 4 0.0060 4 0.0425 4 0.0346 4 0.0362 4 0.0961 4 0.1460 4 0.1657 4 0.0517 4 0.0517 4 0.0517 4 0.0821 4 0.5129 4 0.0060 4 0.1230 4 0.1277 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 low UOA 0 0 0 0 0 0 0 0 000147 1995-.08-23 11:00 output of : 950823Al Fluoride 1.5 Fluoride L PPM ppm Poo Typ Ident Ch Result F Time Ch Result F Time 54 u SPK 250-2 3 0.218 9490 4 0.2034 0 55 u SPK 250-3 3 0.291 9666 4 0.2496 0 56 u SPK 250-4 3 0.253 9839 4 0.2269 0 57 u BLK 3 0.092 10014 4 0.0651 0 58 u BLK 3 0.087 10189 4 0.0563 0 59 u F52712-2 3 0.083 10365 4 0.0477 0 60 u F52729-2 3 0.075 10535 4 0.0310 0 61 u F52803-2 3 0.075 10715 4 0.0316 0 62 d Drift 3 1.522 10891 4 0.5121 0 63 w Wash 3 0.064 11131 4 0.0060 0 64 u F52716-2 3 0.075 11238 4 0.0324 0 65 u F52728-2 3 0.078 11413 4 0.0384 0 66 u F52787-2 3 0.076 11592 4 0.0341 0 67 u F52717-2 3 0.075 11764 4 0.0310 0 68 u SPK 62-1 3 0.083 11941 4 0.0487 0 69 u SPK 62-2 3 0.109 12117 4 0.0914 0 70 u SPK 250-1 3 0.117 12292 4 0.1030 0 71 u SPK 250-2 3 0.238 12468 4 0.2170 0 72 u SPK 250-3 3 0.182 12645 4 0.1746 0 73 d Drift 3 1.537 @12817 4 0.5137 0 74 w Wash 3 0.064 13048 4 0.0060 0 wt rw RunOut Wash 3 0.064 13292 4 0.0060 0 Page 2 of 2 000148 Calibration curve of SS0823AI : Fluoride L 0.2286473- I-TB 6 nw.TaTa-ziiaw-iUmTo0g 007984S 64 0. DOSS961 0 Order 2 Measured 900 r 0.99897 000149 1.63733461 Calibration curve of 950823AI Fluoride 1.5 0.06446440MMM6-4-= 0.0637903 0 Order Measured Inverse Logarithm 409i s 6330 000150 4095 ,7,7. Raw data of 950823AI : Fluoride I.S 4 -7 0 0 Esc=Exit FI=Help T i'me Crtl-P=Edit peaks 1 soo'o 000151 409S F4ogFs zy Raw data of 950823Al : Fluoride i.S 4560 64M z-9 f@ 12--53 If yy3r -57 ;1qoql 0 4!;60 Time Esc=Exit : FI=Help 1 Crtl-P=Edit peaks 4 qf kki @ @j 9560 000152 ,so Raw data of 950823AI : Fluoride t.5 294 64 4OSS 55 S, 0 89,35 Esc=Exit FI=Help le A Q Time Crtl-P=Edit peaks 1 4;470 13935 0001!i,3 1995-08-23 15:23 output of : 950823BI Software : version 6.1 cl990,93 operator DDW Date of the Analysis : 1995-08-23 10:50 Analysis File Name : C:\SKALAR\DATA\HWIDATA\SERUM\950823BI II0-k Fluoride 1.5 Calibration order = Inverse Logarithm Slope 5 = #.#off# x - cl Result = 101 1 s I x = corrected value of the sample cl = corrected value of the concentration 1 s = Slope of the electrode a2 = al = aO = -0.00000 0.00070 -1.18413 Fluoride L Calibration order = 2 Correlation r = 0.99924 Result = a2 x2 + al x + aO a2 = al-= aO -0.00000 0.00027 0.00378 Sampler Type Number Sample Time Wash Time Air Time Take up special needle Height : SA1000 :1 : 50 sec. : 120 sec. : I sec. : Single : None : 70 mm. Diluter needle Height dilution Factor dilution Volume Resample Dilution runs 80 mm 10 2.5 ml. 1 1 User file : TXT Reproces : No 000154. 1995-08-23 15:23 output of 950823Bl Flu6ride 1.5 Path number Signal type Decolor system Number diluto Resample dil Threshold diG output Window event 3 : Debubbled : yes :0 : No : No : 4095 :0 : Off 81 standard : Ignore 92 standard : Ignore s3 standard : Ignore s4 standard : Ignore ani@ar : Ign S% gs@lan arri : .150 s7 standard : 0.300 S8 standard : 0.600 89 standard : 1.200 slO standard : 1.500 Order : Inverse Logarithm Dimension : PPM start Value : 500 DU trigger Limit : 1800 Sec Peak shape : Pointed start ignore : .60 Sec aNd ignore : i20 Sec Measure window : 75 0 Filter : No Regeneration : No formula output ##.$so Fluoride L Path number Signal type Decolor system Number diluto Resample dil Threshold diG output Window event :0 : Debubbled : No :0 : No : No : 4095 :0 : Off 000155 1995-08-23 15:23 OutPut of sl standard : 0.015 s2 standard : 0.030 s3 standard : 0.060 s4 standard : s5 standard : 0.090 0.120 86 standard : 0.150 s7 standard : Ignore S8 standard : Ignore 89 standard : Ignore slO standard : Ignore Order : 2 Dimension : PPM start Value : 500 DU trigger Limit : 1800 Sec Peak shape : Pointed start ignore : 60 Sec eNd ignore : 120 Sec Measure window : 75 Filter : No Regeneration : No formula : c4:=c3 output : 0.8### 950823BI OOOJLSG 1995-08-23 15:23 C)UtPut of : 950823BI Fluoride 1.5 Fluoride L PPM PPM Pos Typ ident Ch Result F Time Ch Result F Time wt iw 1t 2d 3w 4 sl 5 s2 6 s3 7 s4 8 s5 9 s6 10 s7 11 88 12 s9 13 810 14 d 15 w 16 u 17 u 18 u 19 u 20 u 21, u 22 u 23 u 24 u 25 u 26 d 27 w 28 u 29 u 30 u 31 u 32 u 33 u 34 u 35 u 36 u 37 u 38 d 39 w 40 u 41 u 42 u 43 u 44 u 45 u 46 u 47 u 48 u 49 u 50 d 51 w 52 u 53 u Initial Wash 3 0.065 Tracer 3 1.476 Drift 3 1.484 Wash 3 0.065 Standard 1 3 0.070 Standard 2 3 0.076 Standard 3 3 0.092 Standard 4 3 0.111 Standard 5 3 0.129 Standard 6 3 0.157 Standard 7 3 0.277 Standard 8 3 0.621 Standard 9 3 1.225 Standard 10 3 1.470 Drift 3 1.489 Wash SERUM BLK SERUM BLK SERUM BLK 3 0.065 3 0.087 3 0.084 3 0.078 SPK 62-1 3 0.087 SPK 62-2 3 0.119 SPK 62-3 3 0.121 SPK 250-1 3 0.114 SPK 250-2 3 0.233 SPK 250-3 3 0.208 SPK 124-1 3 0.204 Drift 3 1.468 Wash 3 0.065 SPK 124-2 3 0.206 BLK 3 0.136 BLK 3 0.090 P52726-2 F52735-2 3 0.082 3 0.080 F52732-2 F52736-2 F52739-2 F52711-4 3 0.079 3 0.076 3 0.076 3 0.075 F52721-4 3 0.074 Drift 3 1.504 Wash F52719-4 F52727-4 F52725-4 F52723-4 SPK 124-1 3 0.065 3 0.255 3 0.079 3 0.077. 3 0.093 3 0.165 BLK 3 0.083 BLK 3 0.082 SPK 62-1 3 0.090 SPK 62-2 3 0.105 SPK 62-3 3 0.118 Drift 3 Wash 3 SPK 250-1 3 1.483 0.065 0.171 SPK 250-2 3 0.248 65 212 386 627 730 911 1087 1263 1439 1614 1786 1960 2136 2312 2487 2727 2840 3012 3189 . 3363 3538 3714 3888 4065 4239 4415 4589' 4825 4940 5116 5288 5464 5638 5817 5994 6163 6336 6503 6691 6933 7055 7217 7393 7566 7742 17917 8092 8267 8438 8618 8792 9029 9142 9317 4 0.0038 4 0.5182 4 0.5190 4 0.0038 4 0.0159 4 0.0284 4 0.0604 4 0.0921 4 0.1175 4 0.1507 4 0.2461 4 0.3798 4 0.4895 4 0.5177 4 0.5196 4 0.0038 4 0.0519 4 0.0467 4 0.0334 4 0.0514 4 0.1042 4 0.1066 4 0.0971 4 0.2171 4 0.1981 4 0.1951 4 0.5174 4 0.0038 4 0.1966 4 0.1266 4 0.0583 4 0.0413 4 0.0368 4 0.0347 4 0.0287 4 0.0289 4 0.0263 4 0.0236 4 0.5211 4 0.0038 4 0.2321 4 0.0350 4 0.0302 4 0.0624 4 0.1594 4 0.0444 4 0.0418 4 0.0568 4 0.0837 4 0.1032 4 0.5190 4 0.0038 4 0.1649 4 0.2273 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 o 0 0 0 0 0 0 0 0 0 0 0 0 0 Pags 1 of 2 000157 1995-08-23 15:23 OutPut of : 950823Bl Fluoride 1.5 Fluoride L ppm PPM Pos Typ Ident Ch Result F Time Ch Result F Time 54 u SPK 250-3 3 0.294 9494 4 0.2558 0 55 u BLK 3 0.124 9669 4 0.1115 0 56 u BLK 3 0.102 9841 4 0.0787 0 57 u BLK 3 0.074 10017 4 0.0239 0 58 u F52726-2 3 0.080 10193 4 0.0373 0 59 u F52732-2 3 0.070 10367 4 0.0140 0 60 u F52739-2 3 0.068 10543 4 0.0100 0 61 u F52735-2 3 0.073 10719 4 0.0215 0 62 d Drift 3 1.447 10891 4 0.5152 0 63 w Wash 3 0.065 11082 4 0.0038 0 64 u F52736-2 3 0.076 11245 4 0.0287 0 65 u P52711-4 3 0.063 11433 4 #.##$# 0 66 u BLK 3 0.125 11594 4 0.1127 0 67 u BLK 3 0.182 11769 4 0.1761 0 68 u BLK 3 0.081 11942 4 0.0397 0 69 u BLK 3 0.073 12123 4 0.0212 0 70 u SPK 62-1 3 0.102 12289 4 0.0792 0 71 u SPK 62-2 3 0.105 12469 4 0.0827 0 72 u SPK 250-1 3 0.174 12645 4 0.1685 0 73 u SPK 250-2 3 0.178 12819 4 0.1717 0 74 d Drift 3 1.494 12993 4 0.5200 0 75, w Wash 3 0.065 13222 4 0.0038 0 76 u SPK 250-3 3 0.205 13343 4 0.1953 0 77 u SPX 124-1 3 0.166 13519 4 0.1599 0 78- u SPK 124-2 3 0.160 13693 4 0.1539 0 79 u BLK 3 0.105 13869 4 0.0839 0 80 u BLK 3 0.096 14044' 4 0.0688 0 81 u P52711-4 3 0.084 14214 4 0.0449 0 82 u P52719-4 3 0.079 14396 4 0.0366 0 83 u SPK 124-1 3 0.139 14567 4 0.1311 0 84 d Drift 3 1.471 14743 4 0.5177 0 85 w Wash 3 0.065 14966 4 0.0038 0 wt rw RunOut Wash 3 0.065 15218 4 0.0038 0 Page 2 of 2 000158 Calibration curve of 950823Bl EiioFriT#T-M-3INiM-!Mrr&"Xkg 6 0.21198330, Fluoride L 0-0054010OFiT.31MG@4@M 0.0037783.4( 0 Order 2 Measured 900 r 0.99924 000159 ::@7 t.7792010, Calibration curve of 95082381 Fluoride 1.5 0.06S4SIt Order Measured Inverse Logarithm 409S S ssss OOOIC-0 Raw data of 95082381 : Fluoride 1.5 409S MIFIEM 180 atz-3-tmr,4 @c) 0 0 Esc=Exit FI=Help Ilk, Time Crtl-P=Edit peaks scoo 000161 Raw data of sso823B, : Fluoride I.,r 09. 5 4D P4 "P 0 4560 Esc,@Exit 1 FI=Help Cr lip, % Time 9560 00016Z Raw data of 95082381 : Fluoride 1.5 MOREEM 8935 296 4095 A,4 45 0 89@,r Esc=Exit FI=Help Time Crti-P=Edit Peaks i 4) 139351 oool(;.3 4095 MZFtis mis@= Raw data of 95082381 : Fluoride 1.5 13310 OW-IM-M @7-6 FtT-31*.V6 4 4qi 1 2 PO ru 0 13310 Esc=Exit ; FI=Help Time Crtl-P=Edit peaks 18310 000164 feZ, Software version 6.1 cl990,93 Operator : DDW Dati of the Analysis : 1995-08-28 07:32 Analysis File Name : C:\SKALAR\DATA\HWIDATA\SERUM\95082SAl Fluoride 1.5 Calibration order = Inverse Logarithm Slope s 1.01100 x cl Result = 101 a- 1 x = corrected value of the sample cl = corrected value of the concentration 1 s = Slope of the electrode a2 = al = ao = -0.00000 0.00075 -1.16846 Fluoride L Calibration order = 2 Coz,rolation r = 0.99897 Result = a2 x2 + al * x + aO a2 = al = aO*= -0.00000 0.00032 0.00531 Sampler Type Number Sample Time Wash Time Air Time Take up special needle Height : SA1000 :1 : 50 sec. : 120 sec. : 1 sec. : Single : None : 70 mm. Diluter needle Height : 80 mm dilution Factor : 10 dilution Volume : 2.5 ml. Resample :I Dilution runs : 1 User file : TXT Reproces : No ()OOIGS 1995-08-28 15:03 OutPut Of Fluoride 1.5 Path number Signal type Decolor system Number dilute Resample dil Threshold diG output Window event :3 : Debubbled : Yes :0 : No : No : 4095 :0 : Off sl standard : Ignore s2 standard : Ignore s3 standard : Ignore s4 standard : Ignore s5 standard : Ignore s6 standard : 0.150 s7 standard : 0.300 s8 standard : 0.600 89 standard : 1.200 slO standard : 1.500 Order : Inverse Logarithm Dimension : PPM start Value : 500 DU trigger Limit : 1800 Sac Peak shape start ignore eNd ignore : Pointed : 60 Sac : i20 Sac Measure window : 75 t Filter : No Regeneration : No formula output Fluoride L Path number Signal type Decolor system Number dilute Resample dil Threshold diG output Window event 0 Debubbled No 0 No No 4095 0 Off 95082SAl 000166 1995-08-28 15:03 output of 81 standard : 0.015 s2 standard : 0.030 s3 standard : 0.060 s4 standard : 0.090 s5 standard : 0.120 96 standard : 0.150 s7 standard : Ignore ss standard : Ignore S9 standard : Ignore slO standard : Ignore Order : 2 Dimension : PPM start Value : 500 DU trigger Limit : 1800 Sec Peak shape : Pointed start ignore : 60 Sec eNd ignore : 120 Sec Measure window : 75 t Filter : No Regeneration : No formula c4:=c3 output 95082SAl 000167 4rl' 1995-08-28 15:03 output of : 950828AI Fluoride 1.5 Fluoride L PPM PPM Pos*Typ Ident Ch Result F Time wt iw Initial Wash 3 0.068 1t Tracer 3 1.453 2d Drift 3 1.471 3w Wash 3 0.068 4 sl Standard 1 3 0.071 5 s2 6 s3 Standard 2 3 0.078 Standard 3 3 0.092 7 s4 Standard 4 3 0.109 8 s5 Standard 5 3 0.129 9 s6 Standard 6 3 0.156 10 s7 Standard 7 3 0.279 11 Be Standard 8 3 0.616 12 s9 Standard 9 3 1.233 13 slO Standard 10 3 1.467 14 d Drift 3 1.485 15 w Wash 3 0.068 16 u SERUM BLANK 3 0.091 17 u SERUM BLANK 3 0.091 18 u SPK 62-1 3 0.113 19 u SPK 62-2 3 0.107 20 u SPR 250-1 3 0.195 21 u SPK 250-2 3 0.214 22 u SPK 124-1 3 0.192 23 u SPK 124-2 3 0.176 24 u BLANK 3 0.092 25 u F52725-4 3 0.095 26 d Drift 3 1.462 27 w Wash 3 0.068 28 u F52721-4 3 0.092 29 u F52727-4 3 0.085 30 u F52806-4 3 0.08@ 31 u F52713-4 3 0.079 32 u F52718-4 3 0.087 33 u F52723-4 3 0.081 34 u P52715-4 3 0.080 35 u F52738-4 3 0.097 36 u SPK 62-1 3 0.085 37 u SPK 62-2 3 0.126 38 d Drift 3 1.482 39 w Wash 3 0.068 40 u SPK 124-1 3 0.152 41 u BLANK-1 3 0.088 42 u BLANK-2 3 0.081. 43 u SPK 62-1 3 0.102 44 u SPK 62-2 3 0.106 45 u SPK 250-1 3 0.302 46 u SPK 250-2 3 0.257 47 u BLK 3 0.092 48 u F52809-4 3 0.080 49 u F52724-4 3 0.079 50 d Drift 3 1.469 51 w 52 u Wash F52737-4 3 0.068 3 0.079 53 u P52801-4 3 0.092 65 209 385 587 733 901 1085 1260 1436 1611 1786 1960 2136 2312 2488 2687 2836 3012 3188 3362 3538 3713 3887 4063 4237 4413 4589 4830 4940 5113 5285 5464 5640 5812 5986 6164 6334 6514 6690 6932 7039 7213 7391 7565 7740 -7915 8091 8263 8435 8614 8791 9031 9138 9315 Ch Result F Time 4 0.0053 0 4 0.2787 0 4 0.2740 0 4 0.0053 0 4 0.0136 0 4 0.0315 0 4 0.0609 0 4 0.0895 0 4 0.1188 0 4 0.1507 0 4 0.2370 0 4 0.3222 0 4 0.3177 0 4 0.2751 0 4 0.2699 0 4 0.0053 0 4 0.0577 0 4 0.0577 0 4 0.0961 0 4 0.0867 0 4 0.1857 0 4 0.1995 0 4 0.1833 0 4 0.1697 0 4 0.0598 0 4 0.0659 0 4 0.2764 0 4 0.0053 0 4 0.0606 0 4 0.0467 0 4 0.0458 0 4 0.0321 0 4 0.0497 0 4 0.0367 0 4 0.0345 0 4 0.0700 0 4 0.0467 0 4 0.1152 0 4 0.2708 0 4 0.0053 0 4 0.1466 0 4 0.0518 0 4 0.0376 0 4 0.0791 0 4 0.0847 0 4 0.2478 0 4 0.2256 0 4 0.0604 0 4 0.0361 0 4 0.0321 0 4 0.2746 0 4 0.0053 0 4 0.0321 0 4 0.0601 0 Page 1 of 2 000168 1995-08-28 15:03 output of : 95082SAl Fluoride 1.5 Fluoride L rpm Pos Typ Ident Ch Result F Time 54 u 55 u 56 u 57 u 58 u 59 u 60 u 61 u 62 d 63 w 64 u 65 u 66 u 67 u 68 u 69 u 70 u 71 u 72 u 73 u 74 d 75 w 76 u 77 u 78 u 79 u 80 u 81 u 82 u 83 u 84 d 85 w wt rw P52720-4 3 P52733-4 3 F52788-4 3 F52730-4 3 F52731-4 3 F52802-4 3 SPK 62-1 3 SPK 62-2 3 Drift 3 Wash 3 SPK 250-1 3 SPK 250-2 3 SPK 124-1 3 BLK 3 BLK 3 F52714-4 3 F52734-4 3 F52774-4 3 F52712-4 3 F52729-4 3 Drift 3 Wash 3 F52803-4 3 F52716-4 3 F52728-4 3 F52787-4 3 SPK 62-1 3 SPK 62-2 3 SPK 124-1 3 SPK 124-2 3 Drift 3 Wash 3 RunOut Wash 3 0.085 0.080 0.075 0.073 0.073 0.077 0.086 0.117 1.492 0.068 0.176 0.235 0.160 0.103 0.093 0.085 0.096 0.085 0.080 0.079 1.488 0.068 0.085 0.081 0.079 0.081 0.095 0.122 0.109 0.154 1.506 0.068 0.068 9484 9666 9840 10016 10192 10366 10536 10715 10892 11096 11241 11415 11591 11769 11941 12113 12291 12461 12641 .12817 12993 13224 13336 13516 13688 13866 14042 14218 14391 14570 14744 14981 15219 PPM Ch Result F Time 4 0.0458 0 4 0.0351 0 4 0.0249 0 4 0.0196 0 4 0.0193 0 4 0.0293 0 4 0.0476 0 4 0.1024 0 4 0.2679 0 4 0.0053 0 4 0.1694 0 4 0.2128 0 4 0.1543 0 4 0.0811 0 4 0.0624 0 4 0.0467 0 4 0.0674 0 4 0.0461 0 4 0.0345 0 4 0.0336 0 4 0.2690 0 4 0.0053 0 4 0.0461 0 4 0.0385 0 4 0.0342 0 4 0.0382 0 4 0.0653 0 4 0.1101 0 4 0.0903 0 4 0.1483 0 4 0.2632 0 4 0.0053 0 4 0.0053 0 Page 2 of 2 00016.19 O.Z314781- Calibration curve of SS082SAI Fluoride L 0.0072A790rl?-JM 94 1+ 0 Order 2 Measured god r 0.99897 000170 1.6003117- Calibration curve of 95082SAI Fluoride 1.5 D. 06784!53 0 Order Measured Inverse Logarithm 4095 S 6651 0001'0"1 40915 Raw data Of 950828AI Fluoride 1.5 0 0 Esc.,=Exit FI=Help Time Crtl-P=Edit Peaks scoo 000172 '76 Raw data of 95082BAl : Fluoride 1.5 "VVM 4560 mopvM-V-2476 4095 P. A)A r' Ap "3 Ali 45160 1 Esc=Exit FI=Help Time Crtl-P=Edit Peaks 9560 000173 Raw data of 95082SAI : Fluoride 1.5 474 IT-E3FU 64 4095 47 A 0 893S Time Esc=Exit : FI=Help Crtl-P=Edit peaks A 13935 000174 Raw data of 9SO81@8AI : Fluoride 1.5 "TVM 13310 426 101PWS-4@-- 4095 PO 1 13310 1 Esc=Exit FI=Help Time Crtl-P=Edit peaks Printo of Sample Table N2946- Inserted in System Table of 0 iw I , ial Wash 1 1.0000 1t racer 1 1.@000 2d 'ft 1 1.000 3w Wa 1 1.000 4 sl Standard 1 1.000 5 s2 Sta@n da rd 2 1.000 6 s3 Standard 3 1.000 7 s4 Standard 4 1 .000 8 S5 Standard 5 1. 00 9 s6 Standard 6 1 1.0 10 s7 Standard 7 1 1.000 11 s8 Standard 8 1 1.000 18310 group 1 000175 1995-08-29 06:38 OutPut of : 95082BB1 Sofiware version 6.1 cl990,93 Operator : DDW Date of the Analysis : 1995-08-28 11:47 Analysis File Name : C:\SKALAR\DATA\HWIDATA\SERUM\95082SB1 Fluoride 1.5 Calibration order = Inverse Logarithm Slope : 5 = 0.010#0 t = lo[ x - cl Reaul s a2 = al = aO = -0.00000 0.00071 -1.18575 x = corrected value of the sample cl = corrected value of the concentration 1 s = Slope of the electrode Fluoride L Calibration order = 2 Co.rrelation z-= 0.99313 Result a2 a1 aO a2 x2 + al 0.00000 0.00020 0.00438 x + aO Sampler Diluter Type Number Sample Time Wash Time Air Time Take up special needle Height : SA1000 :1 : 50 sec. : 120 sec. : 1 sec. : Single : None : 70 mm. needle Height : 80 mm dilution Factor : 10 dilution Volume : 2.5 ml. Resample :1 Dilution runs : 1 User file : TXT Reproces : No Sri 9 s 000176 71 1995-08-29 06:38 OutPut of Fluoride 1.5 Path number Signal type Decolor system Number dilute Resample dil Threshold diG output Window event 3 Debubbled Yes 0 No No 4095 0 Off sl standard s2 standard s3 standard s4 standard S5 standard S6 standard s7 standard s8 standard S9 standard S10 standard Order : Inverse Dimension : PPM start Value trigger Limit Peak shape start ignore eNd ignore Measure window Filter Regeneration formula output Ignore Ignore Ignore Ignore Ignore 0.150 0.300 0.600 1.200 1.500 Logarithm : 500 DU : 1800 Sec : Pointed : 60 Sec : 120 Sec : 75 t : No : No Fluoride L Path number Signal type Decolor system Number dilute Resample dil Threshold diG output Window event :0 : Debubbled : No :0 : No : No : 4095 :0 : Off 950828BI 000177 1995-08-29 06:38 output of : 95082SB1 1 standard : 0.015 :2 st&ndard : 0.030 s3 standard : 0.060 4 standard : 0.090 :5 standard : 0.120 s6 standard : 0.150 s7 standard : Ignore S8 standard : Ignore 89 standard : Ignore slO standard : Ignore Order : 2 Dimension : PPM start Value : 500 DU trigger Limit : 1800 Sec Peak shape : Pointed start ignore : 60 Sec eNd ignore : 120 Sec Measure window : 75 s Filter : No Regeneration : No formula c4:=c3 Output 000178 1995-08-29 06:38 OutPut of : 95082SB1 Fluoride 1.5 Fluoride L PPM ppm Poe Typ Ident Ch Result F Time wt iw Initial Wash 3 1t Tracer 3 2d Drift 3 3w Wash 3 4 91 Standard 1 3 5 s2 Standard 2 3 6 93 Standard 3 3 7 s4 Standard 4 3 8 s5 standard 5 3 9 s6 Standard 6 3 10 s7 Standard 7 3 11 ss Standard 8 3 12 s9 Standard 9 3 13 siO Standard 10 3 14 d Drift 3 15 w Wash 3 16 u SERUM BLK 1 3 17 u SERUM BLK 2 3 Is u SPK 62-1 3 19 u SPK 62-2 3 20 u SPK 124-1 3 21 'u SPK 124-2 3 22 u BLK 3 23 u BLK 3 24 u P52717-4 3 25 u P52735-4 3 26 d Drift 3 27 w Wash 3 28 u P52736-4 3 29 u F52726-4 3 30 u F52732-4 3 31 u P52739-4 3 32 u F52711-8 3 33 u P52719-8 3 34 u BLK 3 35 u SPK 62-1 3 36 u SPK 62-2 3 37 u SPK 124-1 3 38 d Drift 3 39 w Wash 3 40 u SPK 124-2 3 41 u BLK 3 42 u BLK 3 43 u F52725-8 3 44 u F52721-8 3 45 u P52727- 3 46 u F52806-8 3 47 u F52713-8 3 48 u F52718-8 3 49 u P52723-8 3 50 d Drift 3 51 w Wash 3 52 u F52715-8 3 53 u F52738-8 3 0.065 1.464 1.474 0.065 0.069 0.085 0.095 0.113 0.133 0.156 0.280 0.618 1.228 1.470 1.498 0.065 0.086 0.082 0.127 0.118 0.293 0.139 0.097 0.082 0.092 0.081 1.455 0.065 0.080 0.078 0.074 0.072 0.075 0.070 0.079 0.093 0.096 0.108 1.460 0.065 0.174 0.126 0.089. 0.084 0.080 0.077 0.076 0.071 0.072 0.072 1.463 0.065 0.076 0.073 65 212 387 616 727 909 1087 1263 1439 1615 1787 1963 2137 2313 2487 2686 2832 3011 3189 3363 3540 3714 3892 4064 4242 4414 4588 4815 4936 5116 5288 5463 5635 5812 5992 6165 6341 6515 6689 6904 7041 7215 7389 7566 7739 -7914 8092 8266 8439 8613 8789 9028 9133 9313 Ch Result F Time 4 0.0044 0 4 1.5923 0 4 1.6075 0 4 0.0044 0 4 0.0111 0 4 0.0394 0 4 0.0580 0 4 0.0861 0 4 0.1182 0 4 0.1522 0 4 0.3134 0 4 0.6720 0 4 1.2898 0 4 1.6007 0 4 1.6438 0 4 0.0044 0 4 0.0420 0 4 0.0343 0 4 0.1082 0 4 0.0946 0 4 0.3291 0 4 0.1277 0 4 0.0602 0 4 0.0353 0 4 0 0516 0 4 0:0334 0 4 1.5790 0 4 0.0044 0 4 0.0318 0 4 0.0269 0 4 0.0210 0 4 0.0163 0 4 0.0227 0 4 0.0123 0 4 0.0298 0 4 0 0545 0 4 0:0585 0 4 0.0791 0 4 1.5857 0 4 0.0044 0 4 0.1784 0 4 0.1074 0 4 0.0467 0 4 0.0390 0 4 0.0316 0 4 0.0267 0 4 0.0247 0 4 0.0157 0 4 0.0163 0 4 0.0161 0 4 1.5898 0 4 0.0044 0 4 0.0243 0 4 0.0191 0 Page 1 of 2 oool"f4!) 1995-08-29 06:38 OutPut of : 950828Bl Fluoride 1.5 Fluoride L ppm ppm Pos Typ Ident Ch Result F Time Ch Result F Time 54 u P52809-8 3 0.072 9488 4 0.0165 0 55 u SPK 62-1 3 0.088 9662 4 0.0451 0 56 u SPK 62-2 3 0.100 9838 4 0.0648 0 57 u SPK 124-1 3 0.121 10014 4 0.0991 0 58 u SPK 124-2 3 0.136 10190 4 0.1220 0 59 u BLK 3 0.083 10364 4 0.0362 0 60 u BLK 3 0.074 10538 4 0.0197 0 61 u SPK 62-1 3 0.106 10714 4 0.0752 0 62 d Drift 3 1.460 10888 4 1.5857 0 63 w Wash 3 0.065 11129 4 0.0044 0 64 u SPK 62-2 3 0.109 11240 4 0.0799 0 65 u SPK 124-1 3 0.147 11414 4 0.1393 0 66 u SPK 124-2 3 0.149 11592 4 0.1425 0 67 u 68 u BLK 3. 0.089 11765 4 0.0477 0 BLK 3 0.084 11941 4 0.0378 0 69 u F52724-8 3 0.083 12114 4 0.0369 0 70 u P52737-8 3 0.083 12290 4 0.0362 0 71 u F52801-8 3 0.099 12466 4 0.0643 0 72 u F52720-8 3 0.088 12640 4 0.0460 0 73 u F52733-8 3 0.078 .12814 4 0.0269 0 74 d Drift 3 1.465 12988 4 1.5940 0 75 'w Wash 3 0.065 13219 4 0.0044 0 76 u F52788-8 3 0.071 13336 4 0.0154 0 77 u SPK 62-1 3 0.094 13514 4 0.0558 0 78 u SPK 62-2 3 0.100 13692 4 0.0653 0 79 u SPK 124-1 3 0.126 13866 4 0.1074 0 80 d Drift 3 1.468 14039 4 1.5982 0 81 w Wash 3 0.065 14280 4 0.0044 0 wt rw RunOut Wash 3 0.065 14514 4 0.0044 0 Page 2 of 2 ooolso Calibration curve of 95082881 : Fluoride L EN swr%T%7W-I3N if7.rnT7--Ttg6 0.2082849- Sk-orir4-TilgMr.mrmg0 DOSS7780FPT-JIM 64 M 0.0043776. 0 Order Measured 900 r 0.39313 0()Olsl 1.7548071- Calibration curve of 9SO828BI Fluoride I.S -M o.%@EFS m" r,4 0. 06s,-ngo 0 Order Measured Inverse Logarithm 409S s 6008 000182 4095 Raw data of 95082881 : Fluoride 1.5 o E[mrr -m 2RR I- 64 A7 JA 0 0 [Esc=Exit FI=Help Time Crtl-P=Edit peaks scoo ()00183 Raw data of 95082881 : Fluoride l.S 40aS 4F;gn MOM= 2485::@@gl= 0 4@90 E.sc=Exit Fi=Help Time Crtl-P=Edit peaks 1 9590 0()0184 40$S E:FIIE@@@ Raw data of 9SO82881 : Fluoride 1.5 R92r, 448 64 Pd e7i 0 8935 Time Esc=Exit FI=Help 1 Crtl-P=Edit peaks 1 13935 oools5 409S Raw data of 95082881 : Fluoride I.S f!M 12210 SM-IM-M 3B4 94 0 13310 Esc=Exit ; FI=Help T ime Crtl-P=Edit peaks 18310 ()0018G 9.11.4Summary and raw data;ppm F-in serum as determinedby thennalextractiofnollowedby analysis usingOrion ion analyzer. 0()01IS7 HWI 6329-152 AMDT 011095.1 Dohr-mann Serum Analysis AnalysisDates:08/02/95-08/18/95 Allserum sampleswere thermallyextractebdy a modifiedDohrmann DX2000 OrganicHalideAnalyzerand collecteidna 1:1milliQ waterand TISAB solution.The sampleswere measured on an OrionEA940 expandableionanalyzer.'Me Dohrmann was calibrateudsing34ppm, 40ppm, 62ppm, 100ppm, 124ppm, 250ppm, and 500ppm FC-95 standards. T'heOrion was calibratebdy directmeasurement withno blankcorrection using0.05ppm,O.Ippm, 0.5ppm, I.Oppm and 1.5ppm F'standards.The slope,intercepta,nd correlatiownere recordedin theappropriatleogbook. A summary tableisincludeds,howing theppm F- ineach sample (seepage 2).An initiaclalibratiocnurvewithstandarddeviation%,RSD, R 2 value and equationofthelineison paomes 3 - 5. Pages 6 - 19 show theexcelspreadsheethatwas generatedwhen the sampleswere beinganalyzed.The Dohrmann FC-95 initiaclalibration curvewas notused to generatethedatainthespreadsheet.Any Orion readingbelow 0.05 isbelow theOrion calibratioand shouldbe considered an estimate. The FC-95 initiaclalibratiocnurvewas spikedintobovine serum. However, due toproblemswithblankshavinghigherreadingsthan the samples,theblanksamplesand QC check samplesafter08/11/95were takenfrom study# HWI 6329-123rabbit# F52423,F52346, and F52330. This would affectalltheday 4,day 8 samples,and F52726, F52732, F52739, F52736 of theday 2 samples. This studywas discontinueadfterday 8 becausenothingwas found inthe firs3t days of samplinc, Page I of 19 LU92 Group I Dosage: 0 mg/kg HWI 6329-152 Fluoridceoncentratiinornabbisterum(ppmF-) Sample F52711 F5271 9 F52725 F52721 F52727 F52806 Day I 0.593 0.505 0.531 0.443 0.408 0.472 Day 2 0.381 0.337 0.296 0.257 0.526 0.594 Day 4 0.594 0.518 0.691 0.606 0.496 0.459 Day 8 0.484 0.328 0.600 0.556 0.461 0.462 Group 2 Dosage: 4 mg/kg Sample F52713 F5271 8 F52723 F52715 F52738 F52809 Day I 0.994 0.810 0.730 0.749 0.942 1.12 Day 2 0.400 0.402 0.460 0.410 0.382 0.384 Day 4 0.371 0.590 0.414 0.377 0.827 0.391 Day 8 0.341 0.349 0.346 0.344 0.331 0.303 Group 3 Dosage: 16 mg/kg Sample F52724 F52737 F52801 F52720 F52733 F52788 Day I 0.866 0.870 0.761 0.644 0.775 0.337 Day 2 0.369 0.371 0.609 0.464 0.451 0.511 Day 4 0.373 0.827 0.718 0.531 0.436 0.383 Day 8 0.578 0.563 0.856 0.788 0.491 0.259 Group 4 Dosage: 40 mg/kg Sample F52730 F52731 F52802 F52714 F52734 F52774 Day I 0.785 0.772 0.854 0.71 0.772 0.924 Day 2 0.472 0.372 0.482 0.515 0.434 0.434 Day 4 0.348 0.298 0.464 0.580 0.892 0.570 Group 5 Sample Dosage: 10 X 10 fabric F52712 F52729 F52803 F52716 F52728 F52787 Day I 0.934 0.686 0.948 0.964 1.06 0.890 Day 2 0.649 0.457 0.448 0.419 0.549 0.536 Day 4 0.436 0.4S2 0.474 0.411 0.452 0.443 Group 6 Sample Dosage: 10 X 10 fabric F52717 F52735 F52736 F52726 F52732 F52739 Day I 1.27 1.09 0.894 0.725 1.08 0.675 Day 2 0.510 0.472 0.493 0.653 0.337 0.352 Day 4 0.987 0.638 0.503 0.511 0.425 0.328 0()01,S!) STU Sample ID blank-I blank-2 blank-3 blank-4 blank-5 blank-6 blank-7 blank-8 blank-9 blank-10 blank-li blank-12 blank-13 34ppm-1 34ppm-2 34ppm-3 40ppm-1 40ppm-2 40ppm-3 62ppm-1 62ppm-2 62ppm-3 1 ooppm- 1 I00ppm-2 I00ppm-3 124ppm-1 124ppm-2 124ppm-3 Sample Date 7/26/95 7/26195 7/26/95 7/26/95 7/26/95 7/26/95 7/26/95 7/26/95 7/26/95 7/26/95 7/26/95 7/26/95 7/26/95 Actual reading (ppm F-) 0.04392 0.03695 0.03742 0.04193 0.03201 0.03174 0.02625 0.03425 0.04265 0.03228 0.03017 0.03311 0.02693 Sample TISAB mL FC95 Oty finalvol spiked ImL or 9) (mL) 0.1 2.0 0.1 2.0 0.1 2.0 0.1 2.0 0.1 2.0 0.1 2.0 0.1 2.0 0.1 2.0 0.1 2.0 0.1 2.0 0.1 2.0 0.1 2.0 0.1 2.0 onc.FC9 % Actual solution recovery ppm F- fppm) (ug/uo) in sample 0.8784 0.7390 0.7484 0.8386 0.6402 0.6348 0.5250 0.6850 0.8530 0.6456 0.6034 0.6622 0.5386 Mass spiked (ugF-) Mass recovered (ugF-) 0.0878 0.0739 0.0748 0.0839 0.0640 0.0635 0.0525 0.0685 0.0853 0.0646 0.0603 0.0662 0.0539 7/26/95 0.05378 0.1 2.0 7/26/95 0.04430 0.1 2.0 7/26/95 0,04601 0.1 2.0 0.004 0.004 0.004 34 132% 34 109% 34 113% 1.0756 0.8860 0.9202 0.0817 0.0817 0.0817 0.1076 STD 0.0886 %RS 0.0920 AVE 7/26/95 O@04739 0.1 2.0 7/26/95 0.04881 0.1 2.0 7/26/95 0.05178 0.1 2.0 0.004 0.004 0.004 40 99% 40 102% 40 108% 0.9478 0.9762 1.0356 0.0961 0.0961 0.0961 0.0948 STD 0.0976 %RS 0.1036 AVER 7/26/95 0.06585 0.1 2.0 7/26/95 0.06287 0.1 2.0 7/26195 0.06969 0.1 2.0 0.004 0.004 0.004 62 88% 62 84% 62 94% 1.3170 1.2574 1.3938 0.1489 0.1489 0.1489 0.1317 STD 0.1257 %RS 0.1394 AVER 7/26/95 0.08098 0.1 2.0 7/26/95 0.08998 0.1 2,0 7/26/95 0.1047 0.1 2.0 0.004 0.004 0.004 100 67% 100 75% 100 87% 1.6196 1.7996 2.0940 0.2402 0.2402 0.2402 0.1620 STD 0.1800 %RS 0.2094 AVER 7/26/95 0.09999 0.1 2.0 7/26/95 0.1159 0.1 2.0 7/26/95 0.1219 0.1 2.0 0.004 0.004 0.004 124 67% 124 78% 124 82% 1.9998 2.3180 2.4380 0.2978 0.2978 0.2978 0.2000 STD 0.2318 %RS 0.2438 AVER Sample ID 250ppm-1 250ppm-2 250ppm-3 250ppm-4 500ppm-1 500ppm-2 500ppm-3 500ppm-4 Sample Date Actual Sample TISAB mL FC95 onc. FC9 % Actual reading GtY finalvol spiked solution recovery ppm F. (ppm F-I (mL or g)_ (mL) lppm) lug/ug) insample Mass Mass spiked recovered (ugF-) lugF-) 7/26/95 0.1947 0.1 2.0 7/26/95 0,302 0.1 2.0 7/26/95 0.1981 0.1 2.0 7/26/95 0.1972 0.1 2.0 7/26/95 0.3486 0.1 2.0 7/26/95 0.4286 0.1 2.0 7/26/95 0.3745 0.1 2.0 7/26/95 0.3566 0.1 2.0 0.004 0.004 0.004 0.004 0.004 0.004 0.004 0.004 250 65% 250 101% 250 66% 250 66% 500 58% 500 71% 500 62% 500 59% 3.8940 6.0400 3.9620 3.9440 0.6004 0.6004 0.6004 0.6004 0.3894 0.6040 0.3962 0.3944 6.9720 8.5720 7.4900 7.1320 1.2008 1.2008 1.2008 1.2008 0.6972 0.8572 0.7490 0.7132 0.8 0.7 0.6 0.5 0 0.4 0.3 0.2 0.1 0 0 ul SERUM CURVE 2 CARLTON y 0.6013x + 0.0478 R'- 0.9% 0.2 0.4 0.6 0.8 Mass Spiked(ug) Sample ID BLANK BLANK 62PPM-1 62PPM-2 62PPM-3 250PPM-1 250PPM-2 25OPPM-3 BLANK BLANK BLANK BLANK F52713DAY1 F52718DAYI F52723DAYI F52715DAYI F52738DAYI F52509DAYI F52724DAY1 F52737DAY1 F52801DAYI F52720DAYI 62ppm-1 62ppm-2 250ppm-1 250ppm-2 BLANK BLANK SERUM BLANK SERUM BLANK SERUM BLANK SERUM BLANK SERUM BLANK SERUM BLANK Actual Sample TISAB mL FC95 reading Oty finavlol spiked (ppm F-) (mL org) (mL) 0.0512 0.1 2.0 0.0314 0.1 0.0557 0.1 0.0867 0.1 0.0995 0.1 0.0982 0.1 0.240 0.1 0.235 O'l 0.164 0.1 2.0 2.0 0.004 2.0 0.004 2.0 0.004 2.0 0.004 2.0 0.004 2.0 0.004 2.0 0.0878 0.1 2.0 0.0631 0.1 2.0 0.0404 0.1 2.0 0.0497 0.1 2.0 0.0405 0.1 2.0 0.0365 0.1 2.0 0.0374 0.1 2.0 0.0471 0.1 2.0 0.0561 0.1 2.0 0.0433 0.1 2.0 0.0435 0.1 2.0 0.0381 0.1 2.0 0.0322 0.1 2.0 0.0501 0.1 2.0 0.004 0.0838 0.1 2.0 0.004 0.134 0.1 2.0 0.004 0.233 0.1 0.123 0.1 2.0 0.004 2.0 0.0323 0.1 2,0 0.0478 0.1 2.0 0.0524 0,1 2.0 0.0709 0.1 2.0 0.0612 0.1 2.0 0.0542 0.1 2.0 0.0542 0.1 2.0 Conc.-FC95 solution (ppm) 62 62 62 250 250 250 62 62 250 250 % Actual recovery ppm F- (ug/ug) Insample 75% 116% 134% 33% 80% 78% 67% 113% 45% 78% 1.024 0.628 1.11 1.73 1.99 1.96 4.80 4.70 3.27 1.76 1.26 0.807 0.994 0.810 0.730 0.749 0.942 1.12 0.866 0.870 0.761 0.644 1.00 1.68 2.68 4.67 2.46 0.646 0.956 1.05 1.42 1.22 1.08 1.08 Mass spiked (ugF-) 0.149 0.149 0.149 0.600 0.600 0.600 0.149 0.149 0.600 0.600 Mass recovered (ugF-) 0.102 0.0628 0.111 0.173 0.199 0.196 0.480 0.470 0.327 0.176 0.126 0.0807 0.0994 0.0810 0.0730 0.0749 0.0942 0.112 0.0866 0.0870 0.0761 0.0644 0.100 0.168 0.268 0.467 0.246 0.0646 0,0956 0.105 0.142 0.122 0.108 0.108 !ST,(A Sample ID SERUM BLANK SERUM BLANK SERUM BLANK SERUM BLANK SERUM BLANK SERUM BLANK SERUM BLANK SPIKE 62-1 SPIKE 62-2 SPIKE 250-1 SPIKE 250-2 SPIKE 250-3 SPIKE 250-4 BLANK BLANK BLANK BLANK BLANK BLANK F52733-DAYI F52788-DAYI F52730-DAYi F52731-DAYI F52802-DAY1 F52714-DAYI F52734-DAY1 F52774-DAYI F52712-DAYI F52729-DAY1 62-PPM-1 62-PPM-2 62-PPM-3 250-PPM-1 BLANK Actual Sample TISAB mL FC95 reading Oty finavlol spiked (ppm F-Y (mL org) (m 0.0636 0.1 0.0540 0.1 0.0526 0.1 0.0585 0.1 0.0464 0.1 0.0369 0.1 0.0318 0.1 0.0604 0.1 0.0610 0.1 0.177 0.1 0.184 0.1 0.211 0.1 0.204 0.1 0.170 0.1 0.0644 0.1 0.0476 0.1 0.0413 0.1 0.0346 0.1 0.0274 0.1 0.0388 0.1 0,0168 0.1 0.0392 0.1 0.0386 0.1 0.0427 0.1 0.0355 0.1 0.0386 0.1 0.0462 0.1 0.0467 0.1 0.0343 0.1 0.0604 0.1 0.0768 0.1 0.104 0.1 0.226 0.1 0.0983 0-1 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 0.004 2.0 0.004 2.0 0.004 2.0 0.004 2.0 0.004 2.0 0.004 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 0.004 2.0 0.004 2.0 0.004 2.0 0.004 2.0 Conc. FC95 solution ippm) 62 62 250 250 250 250 62 62 62 250 % Actual recovery ppm F- (ug/ug) insample 1.27 1.08 1.05 1.17 0.928 0.737 0.636 81% 1.21 82% 1.22 59% 3.54 61% 3.67 70% 4.22 68% 4.08 3.41 1.29 0.952 0.825 0.692 0.547 0.775 0,337 0.785 0.772 0.854 0.710 81% 103% 139% 75% 0.772 0.924 0.934 0.686 1.21 1.54 2.07 4.53 1.97 Mass spiked (ugF-) 0.149 0.149 0.600 0.600 0.600 0.600 0.149 0.149 0.149 0.600 Mass recovere (ugF 0.127 0.108 0.105 0.117 0.0928 0.0737 0.0636 0.121 0.122 0.354 0.367 0.422 0.408 0.341 0.129 0.0952 0.0825 0.0692 0.0547 0.0775 0.0337 0.0785 0.0772 0.0854 0.0710 0.0772 0.0924 0.0934 0.0686 0.121 0.154 0.207 0.453 0.197 Sample ID F52803-DAY1 F52716-DAYI F52728-DAYI F52787-DAYI F52717-DAYI F52735-DAY1' F52736-DAY1 F52726-DAYI F52732-DAYI F52739-DAYI 62-PPM-1 62-PPM-2 250-PPM-1 250-PPM-2 SERUM BLANK SERUM BLANK SERUM BLANK SERUM BLANK SERUM BLANK SERUM BLANK SERUM BLANK SERUM BLANK SERUM BLANK SERUM BLANK SERUM BLANK SERUM BLANK SERUM BLANK SPIKE 62-1 SPIKE 62-2 SPIKE 62-3 SPIKE 250-1 SPIKE 250-2 SPIKE 250-3 BLANK Actual Sample TISAB mL FC95 reading Qty finavlol spiked (epm F-) (mL org) (mL) 0.0474 0.1 2.0 0.0482 0.1 2.0 0.0529 0.1 2.0 0.0445 0.1 2.0 0.0636 0.1 0.0547 0.1 0.0447 0.1 0.0362 0.1 0.0538 0.1 0.0338 0.1 0.0636 0.1 0.105 0.1 0.286 0.1 0.272 0.1 0.0766 0.1 2.0 2.0 2.0 2.0 2.0 2.0 2.0 0,004 2.0 0.004 2.0 0.004 2.0 0.004 2.0 0.0844 0.1 2.0 0.0673 0.1 2.0 0.0606 0.1 2.0 0.0588 0.1 2.0 0,0621 0.1 0.0616 0.1 0.0571 0.1 0.0469 0.1 0.0586 0.1 0.0422 0.1 0.0358 0.1 0.0363 0.1 0.104 0.1 0.0708 0.1 0.0881 0.1 0.162 0.1 0.267 0.1 0.245 0.1 0.140 0.1 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 0.004 2.0 0.004 2.0 0.004 2.0 0.004 2.0 0.004 2.0 0.004 2.0 Conc.FC95 solution (PPM) 62 62 250 250 62 62 62 250 250 250 % Actual recovery ppm F- (ug/ug) insample 0.948 0.964 1.06 0.890 1.27 1.09 0.894 0.725 1.08 0.675 85% 1.27 141% 2.09 95% 5.73 91% 5.44 1.53 1.69 1.35 1.21 1.18 1.24 1.23 1.14 0,937 1.17 0.844 0.716 0.725 139% 2.07 95% 1.42 118% 1.76 54% 3.24 89% 5.34 81% 4.89 2.81 Mass spiked (ugF-) 0.149 0.149 0.600 0.600 0.149 0.149 0.149 0.600 0.600 0.600 Mass recovere( (ugF-) 0.0948 0.0964 0.106 0.0890 0.127 0.109 0.0894 0.0725 0.108 0.0675 0.127 0.209 0.573 0.544 0.153 0.169 0.135 0.121 0.118 0.124 0.123 0.114 0.0937 0.117 0.0844 0.0716 0,0725 0.207 0.142 0.176 0.324 0.534 0.489 0,281 5TtAt Sample ID BLANK BLANK F5271 I-DAYI F52719-DAY1 F52725-DAYI F52721-DAY1 F52727-DAY1 F52806-DAYI F5271 I-DAY2 F52719-DAY2 F52725-DAY2 F52721-DAY2 SPIKE 62-1 SPIKE 250-1 SERUM BLANK SERUM BLANK SERUM BLANK SERUM BLANK SERUM BLANK SERUM BLANK SERUM BLANK SERUM BLANK SERUM BLANK SPIKE 62-1 SPIKE 62-2 SPIKE 62-3 SPIKE 62-4 SPIKE 250-1 SPIKE 250-2 SPIKE 250-3 BLANK BLANK BLANK BLANK Actual Sample reading Gly (ppm F-)_(mLor 0.0564 0.1 0,0406 0.1 0.0296 0.1 0.0253 0.1 0.0266 0.1 0.0222 0.1 0.0204 0.1 0.0236 0.1 0.0191 0.1 0.0169 0.1 0.0148 0.1 0.0128 0.1 0.0922 0.1 0.281 0.1 0,0668 0.1 0.0515 0.1 0,0787 0.1 0.0452 0.1 0.0484 0.1 0.0490 0.1 0.0526 0.1 0.0381 0.1 0.0407 0.1 0.052a 0.1 0.0742 0.1 0.0688 0.1 0.0842 0.1 0.149 0.1 0.258 0.1 0.241 0.1 0,137 0.1 0.0685 0.1 0.0492 0.1 0.0394 0.1 TISAB mL FC95 finavlol spiked (mL) 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 0.004 2.0 0.004 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 0.004 2.0 0.004 2.0 0.004 2.0 0.004 2.0 0.004 2.0 0.004 2.0 0.004 2.0 2.0 2.0 2.0 Cone.FC95 solution (ppm) 62 250 62 62 62 62 250 250 250 % Actual recovery ppm F- (ug/ug) insample 1.13 0.812 0.593 0.505 0.531 0.443 0.408 0.472 0.381 0.337 0.296 0.257 124% 1.84 94% 5.62 1.34 1.03 1.57 0.904 0.967 0.979 1.05 0.761 0.814 71% 1.06 100% 1.48 92% 1.38 113% 1.68 49% 2.97 86% 5.15 80% 4.82 2.73 1.37 0.984 0.787 Mass spiked (ugF-) 0.149 0.600 0.149 0.149 0.149 0.149 0.600 0.600 0.600 Mass recovered (ugF-) 0.113 0.0812 0.0593 0.0505 0.0531 0.0443 0.0408 0.0472 0.0381 0.0337 0.0296 0.0257 0.184 0.562 0.134 0.103 0.157 0.0904 0.0967 0.0979 0.105 0.0761 0.0814 0.106 0.148 0.138 0.168 0.297 0.515 0.482 0.273 0.137 0.0984 0.0787 Sample GENNON ID BLANK BLANK F52727 DAY2 F52606 DAY2 F52713 DAY2 F52718 DAY2 F52723 DAY2 F52715 DAY2 F52738 DAY2 F52809 DAY2 F52724 DAY2 F52737 DAY2 SPIKE 62-1 SPIKE 62-2 SPIKE 250-1 SPIKE 250-2 BLANK SERUM BLANK SERUM BLANK SERUM BLANK SERUM BLANK SERUM BLANK SPIKE 62-1 SPIKE 62-2 SPIKE 62-3 SPIKE 250-1 SPIKE 250-2 SPIKE 250-3 SPIKE 250-4 SPIKE 250-5 SPIKE 250-6 BLANK BLANK BLANK Actual @Sample TISAB mL FC95 reading Oty finavlol spiked (ppm F-) (mL org) (mL) 0.0407 0.1 0.0310 0.1 0.0263 0.1 0.0297 0.1 0.0200 0.1 0.0201 0.1 0.0230 0.1 0.0205 0.1 0.0191 0.1 0.0192 0.1 0.0185 0.1 0.0186 0.1 0.0389 0.1 0.0546 0.1 0.118 0.1 0.223 0.1 0.103 0.1 0.0522 0.1 0.0474 0.1 0.0420 0.1 0.0330 0.1 0.0299 0.1 0.0517 0.1 0.0747 0.1 0.0763 0.1 0.112 0.1 0.179 0.1 0.197 0.1 0.194 0.1 0.223 0.1 0.330 0.1 0.102 0.1 0.0411 0.1 0.0185 0.1 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 0.004 2.0 0.004 2.0 0.004 2.0 0.004 2.0 2.0 2.0 2.0 2.0 2.0 2.0 0.004 2.0 0.004 2.0 0.004 2.0 0.004 2.0 0.004 2.0 0.004 2.0 0.004 2.0 0.004 2.0 0.004 2.0 2.0 2.0 Conc. FC95 solution (ppm) 62 62 250 250 62 62 62 250 250 250 250 250 250 % Actual recovery ppm F- (ug/ug) Insample 0.814 0.619 0.526 0.594 0.400 0.402 0.460 0.410 0.382 0.384 0.369 0.371 52% 0.777 73% 1.09 39% 2.37 74% 4.45 2.06 1.04 0.948 0.840 0.659 0.599 69% 1.03 100% 1.49 102% 1.53 37% 2.23 60% 3.57 66% 3.94 65% 3.89 74% 4.46 110% 6.60 2.04 0.822 0.370 Mass Mass spiked recover (ug_F-) lug L-1 0.0814 0,0619 0,0526 0.0594 0.0400 0.0402 0.0460 0.0410 0.0382 0.0384 0.0369 0.0371 0.149 0.0777 0.149 0.109 0.600 0.237 0.600 0.445 0.206 0.104 0.0948 0.0840 0.0659 0.0599 0.149 0.103 0.149 0.149 0.149 0.153 0.600 0.223 0.600 0.357 0.600 0.394 0.600 0.389 0.600 0,446 0.600 0.660 0.204 0.0822 0.0370 CP 5CLAby Sample ID F52801 -DAY2 F52720-DAY2 F52733-DAY2 F52788-DAY2 F52730-DAY2 F52731-DAY2 F52802-DAY2 F52714-DAY2 F52734-DAY2 F52774-DAY2 SPIKE 62-1 SPIKE 250-1 SPIKE 250-2 SERUM BLANK SERUM BLANK SERUM BLANK SERUM BLANK SERUM BLANK SERUM BLANK SERUM BLANK SERUM BLANK SERUM BLANK SPIKE 62-1 SPIKE 62-2 SPIKE 62-3 SPIKE 250-1 SPIKE 250-2 SPIKE 250-3 SPIKE 250-4 BLANK BLANK BLANK BLANK BLANK Actual Sample TISAB mLFC95 reading Qty finalvol spiked (ppm F-)_(mL or g) (mL) 0.0304 0.1 2.0 0.0232 0.1 2.0 0.0225 0.1 2.0 0.0255 0.1 2.0 0.0236 0.1 2.0 0.0186 0.1 2.0 0.0241 0.1 2.0 0.0258 0.1 2.0 0.0217 0.1 2.0 0.0217 0.1 2.0 0.0673 0.1 2.0 0.004 0.115 0.1 2.0 0.004 0.215 0.1 0.0910 0.1 2.0 0.004 2.0 0.0568 0.1 2.0 0.0451 0.1 2.0 0.0438 0.1 2.0 0.0467 0.1 2.0 0.0435 0.1 2.0 0.0436 0.1 2.0 0.0335 0.1 2.0 0.0352 0.1 2.0 0.0395 0.1 0.0631 0.1 2.0 0.004 2.0 0.004 0.0926 0.1 2.0 0.004 0.0998 0.1 2.0 0.004 0.198 0.1 2.0 0.004 0.256 0.1 2.0 0.004 0.217 0.1 2.0 0.004 0.180 0.1 2.0 0.0994 0.1 2.0 0.0783 0.1 2.0 0.0525 0.1 2.0 0,0465 0.1 2.0 Cone.FC95 % Actual solution recovery ppm F- (ppm) (ug/ug) insample 0.609 0.464 0.451 0.511 0.472 0.372 0.482 0.515 0.434 0.434 62 90% 1.35 250 38% 2.30 250 72% 4.31 1.82 1.14 0.903 0.877 0.934 0.870 0.872 0.670 0.704 62 53% 0.790 62 85% 1.26 62 124% 1.85 250 33% 2.00 250 66% 3.96 250 85% 5.13 250 72% 4.34 3.60 1.99 1.57 1.05 0.930 Mass spiked (ugF-) 0.149 0.600 0.600 0.149 0.149 0.149 0.600 0.600 0.600 0.600 Mass recovere( (ugF-) 0.0609 0.0464 0.0451 0.0511 0.0472 0.0372 0.0482 0.0515 0.0434 0.0434 0.135 0.230 0.431 0.182 0.114 0.0903 0.0877 0.0934 0.0870 0.0872 0.0670 0.0704 0.0790 0.126 0.185 0.200 0.396 0.513 0.434 0.360 0.199 0.157 0.105 0.0930 Sample ID BLANK@ BLANK F52712 DAY2 F52729 DAY2 F52803 DAY2 F52716 DAY2 F52728 DAY2 F52787 DAY2 F52717 DAY2 SPIKE 62-1 SPIKE 62-2 SPIKE250-1 SPIKE250-2 SPIKE250-3 SERUM BLANK SERUM BLANK SERUM BLANK SERUM BLANK SERUM BLANK SERUM BLANK SERUM BLANK SPIKE 62-1 SPIKE 62-2 SPIKE 62-3 SPIKE 62-4 SPIKE 62-5 SPIKE 250-1 SPIKE 250-2 SPIKE 250-3 SPIKE 250-4 SPIKE 250-5 SPIKE 250-6 SPIKE 124-1 SPIKE 124-2 Actual Sample reading Oty (PPM F-) (mL org) 0.0417 0.1 0.0385 0.1 0.0325 0.1 0.0229 0.1 0.0224 0.1 0.0209 0.1 0.0275 0.1 0.0268 0.1 0.0255 0.1 0.0356 0.1 0.0697 0.1 0.0820 0.1 0.221 0.1 0.182 0.1 0.114 0.1 0.0836 0.1 0.0620 0.1 0.0521 0.1 0.0426 0.1 0.0375 0.1 0.0341 0.1 0.0471 0.1 0.0592 0.1 0.0481 0.1 0.0574 0.1 0.0469 0.1 0.0871 0.1 0.167 0.1 0.184 0.1 0.187 0.1 0.162 0.1 0.196 0.1 0.171 0.1 0.0473 0.1 TISAB finavlol (mL) 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 mL FC95 spiked 0.004 0.004 0.004 0.004 0.004 0.004 0.004 0.004 0.004 0.004 0.004 0.004 0.004 0.004 0.004 0.004 0.004 0.004 Conc. FC95 solution (ppm)-_ 62 62 250 250 250 62 62 62 62 62 250 250 250 250 250 250 124 124 % Actual recovery ppm F- (ug/ug)-insample 48% 94% 27% 74% 60% 63% 79% 65% 77% 63% 29% 56% 61% 62% 54% 65% 115% 32% 0.834 0.770 0.649 0.457 0.448 0.419 0.549 0.536 0.510 0.712 1.39 1.64 4.41 3.63 2.28 1.67 1.24 1.04 0.852 0.751 0.682 0.943 1.18 0.963 1.15 0.939 1.74 3.33 3.69 3.73 3.24 3.91 3.42 0.946 Mass spiked (ugF-) 0.149 0.149 0.600 0.600 0.600 0.149 0.149 0.149 0.149 0.149 0.600 0.600 0.600 0.600 0.600 0.600 0.298 0.298 Mass recovered (ugF-) 0.0834 0.0770 0.0649 0.0457 0.0448 0.0419 0.0549 0.0536 0.0510 0.0712 0.139 0.164 0.441 0.363 0.228 0.167 0.124 0.104 0.0852 0.0751 0.0682 0.0943 0.118 0.0963 0.115 0.0939 0.174 0.333 0.369 0.373 0.324 0.391 0.342 0.0946 rj Sample ID SPIKE 124-3 SPIKE 124-5 SPIKE 124-6 SERUM BLANK SERUM BLANK SERUM BLANK SERUM BLANK SERUM BLANK SERUM BLANK SPIKE 62-1 SPIKE 62-2 SPIKE 62-3 SPIKE 250-1 SPIKE 250-2 SPIKE 250-3 SPIKE 124-1 SPIKE 124-2 BLANK BLANK F52726 DAY 2@ F52735 DAY 2*** F52732 DAY 2@* F52736 DAY 2"* F52739 DAY 2*** F52711 DAY 4*** F52721 DAY 4*** F52719 DAY 4*** F52727 DAY 4*** F52725 DAY 4*** SPIKE 62-1 SPIKE 124-1 BLANK BLANK 62PPM-1 Actual Sample TISAB mL FC95 reading oty finavlol spiked (ppm F-) (mL org) (mL) 0.0133 0.1 0.161 0.1 0.155 0.1 2.0 0.004 2.0 0.004 2.0 0.004 0.143 0.1 2.0 0.0822 0.1 2.0 0,0677 0.1 2.0 0.0383 0.1 2.0 0.0366 0.1 0.0272 0.1 0.0431 0.1 0.0855 0.1 0.0877 0.1 0.0769 0.1 0.217 0.1 0.186 0.1 0.179 0.1 0.182 0.1 0,0966 0.1 2.0 2.0 2.0 0.004 2.0 0.004 2.0 0.004 2.0 0.004 2.0 0.004 2.0 0.004 2.0 0.004 2.0 0,004 2.0 0.0395 0.1 2.0 0.0264 0.1 2.0 0.0225 0.1 2.0 0.0208 0.1 2.0 0.0179 0.1 2.0 0.0177 0.1 2.0 0.0164 0.1 2.0 0.0146 0.1 2.0 0.0171 0.1 2.0 0.0168 0.1 0.0170 0.1 0.0407 0.1 0.130 0.1 2.0 2.0 2.0 0.004 2.0 0.004 0.0311 0.1 2.0 0.0291 0.1 0.0407 0.1 2.0 2.0 0.004 Conc. FC95 solution (ppm) 124 124 124 62 62 62 250 250 250 124 124 62 124 62 .% Actual recovery ppm F- (uglug) insamele 9% 108% 104% 58% 115% 118% 26% 72% 62% 120% 122% 0.265 3.21 3.09 2.85 1.64 1.35 0.767 0.732 0.545 0.863 1.71 1.75 1.54 4.35 3.72 3.58 3.64 1.93 0.790 0.529 0.450 0.416 0.357 0.354 0.328 0.293 0.342 0.335 0.341 55% 0.813 88% 2.61 0.623 0.582 55% 0.814 Mass spiked lug F-) 0.298 0.298 0.298 0.149 0.149 0.149 0.600 0.600 0.600 0.298 0.298 0.149 0.298 0.149 Mass recovere( (ugF-) 0.0265 0.321 0.309 0.285 0.164 0.135 0.0767 0.0732 0.0545 0.0863 0.171 0.175 0.154 0.435 0.372 0.358 0.364 0.193 0.07,90 0.0529 0.0450 0.0416 0.0357 0.0354 0.0328 0,0293 0,0342 0.0335 0.0341 0.0813 0.261 0.0623 0.0582 0.0814 Sample ID 62PPM-2 62PPM-3 25OPPM -1 25OPPM -2 25OPPM -3 BLANK BLANK BLANK F52726 DAY2 F52732 DAY2 F52739 DAY2 F52735 DAY2 F52736 DAY2 F52711 DAY2 BLANK BLANK BLANK BLANK SPIKE 62-1 SPIKE 62-2 SPIKE 250-1 SPIKE 250-2 SPIKE 250-3 SPIKE 124-1 SPIKE 124-2 BLANK BLANK F52711 DAY4 F52719 DAY4 SPIKE 62-1 SPIKE 124-1 BLANK BLANK BLANK Actual Sample reading Oty (ppm F-) (mL org) 0.0613 0.1 0.0798 0.1 0.141 0.1 0.223 0.1 0.267 0.1 0.0865 0.1 0.0582 0.1 0.0213 0.1 0.0326 0.1 0.0169 0.1 0.0176 0.1 0.0236 0.1 0.0246 0.1 0.0064 0.1 0.0888 0.1 0.150 0.1 0.0251 0.1 0.0175 0.1 0.0556 0.1 0.0570 0.1 0.133 0.1 0.138 0.1 0.159 0.1 0.122 0.1 0.121 O'l 0.0566 0.1 0.0457 0.1 0.0297 0.1 0.0259 0.1 0.0563 0.1 0.125 0.1 0.0673 0.1 0.0302 0.1 0.0334 0.1 TISAB finavlol (mL) 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 mL FC95 spiked 0.004 0.004 0.004 0.004 0.004 0.004 0.004 0.004 0.004 0.004 0.004 0.004 0.004 0.004 Conc.FC95 solution (ppm) 62 62 250 250 250 62 62 250 250 250 124 124 62 124 % Actual recovery ppm F- (ug/ug) Insample 82% 1.23 107% 1.60 47% 2.81 74% 4.45 89% 5.35 1.73 1.16 0.426 0.653 0.337 0.352 0.472 0.493 0.128 1.78 3.00 0.501 0.350 75% 1.11 77% 1.14 44% 2.66 46% 2.76 53% 3.18 82% 2.43 81% 2.42 1.14 0.913 0.594 0.518 76% 1.13 84% 2.51 1.35 0.603 0,668 Mass spiked _(ugF-) 0.149 0.149 0.600 0.600 0.600 0.149 0.149 0.600 0.600 0.600 0.298 0.298 0.149 0.298 Mass recovere( (ugL-L 0.123 0.160 0.281 0.445 0.535 0.173 0.116 0.0426 0.0653 0.0337 0.0352 0,0472 0.0493 0.0128 0.178 0.300 0.0501 0.0350 0.111 0.114 0.266 0.276 0.318 0.243 0.242 0.114 0.0913 0.0594 0.0518 0.113 0.251 0.135 0.0603 0.0668 !5rLAbl Sample ID SPIKE 62-1 SPIKE 62-2 SPIKE 62-3 SPIKE 250-1 SPIKE 250-2 SPIKE 250-3 SPIKE 250-4 SPIKE 250-5 SPIKE 124-1 SPIKE 124-2 BLANK BLANK F52725 DAY 4 F52721 DAY 4 F52727 DAY 4 F52806 DAY 4 F52713 DAY 4 F52718 DAY 4 F52723 DAY 4 F52715 DAY 4 F52738 DAY 4 SPIKE 62-1 SPIKE 62-2 SPIKE 124-1 SERUM BLANK SERUM BLANK SERUM BLANK SPIKE 62-1 SPIKE 62-2 SPIKE 62-3 SPIKE 250-1 SPIKE 250-2 SPIKE 250-3 SPIKE 250-4 Actual Sample TISAB mL FC95 reading Oty finalvol spiked (ppm _F-)(mL org) (mL) 0.0317 0.1 0.0656 0.1 0.0567 0.1 0.0942 0.1 0.171 0.1 0.180 0.1 0.158 0.1 0.172 0.1 0.153 0.1 0.129 0.1 0.0706 0.1 2.0 0.004 2.0 0.004 2.0 0.004 2.0 0.004 2.0 0.004 2.0 0.004 2.0 0.004 2.0 0.004 2.0 0.004 2.0 0.004 2.0 0.0293 0.1 2.0 0.0346 0.1 2.0 0.0303 0.1 2.0 0.0248 0.1 2.0 0.0230 0.1 2.0 0.0185 0.1 2.0 0.0295 0.1 2.0 0.0207 0.1 2.0 0.0189 0.1 2.0 0.0414 0.1 0.0244 0.1 0.0784 0.1 0.126 0.1 2.0 2.0 0.004 2.0 0.004 2.0 0.004 0.0334 0.1 2.0 0.0290 0.1 2.0 0.0239 0.1 0.0414 0.1 0.0534 0.1 0.0564 0.1 0.127 0.1 0.223 0.1 0.172 0.1 0.264 0.1 2.0 2.0 0.004 2.0 0.004 2.0 0.004 2.0 0.004 2.0 0.004 2.0 0.004 2.0 0.004 Conc.FC95 solution (ppm) 62 62 62 250 250 250 250 250 124 124 62 62 124 62 62 62 250 250 250 250 % Actual recovery ppm F(ug/ug) Insample 43% 0.635 88% 1.31 76% 1.13 31% 1.88 57% 3.42 60% 3.60 53% 3.16 57% 3.44 103% 3.06 87% 2.58 1.42 0.585 0.691 0.606 0.496 0.459 0.371 0.590 0.414 0.377 0.827 33% 0.487 105% 1.57 85% 2.52 0.668 0.580 0.478 56% 0.827 72% 1.07 76% 1.13 42% 2.54 74% 4.45 57% 3.45 88% 5.29 Mass spiked (ugF-) 0.149 0.149 0.149 0.600 0.600 0.600 0.600 0.600 0.298 0.298 0.149 0.149 0.298 0.149 0.149 0.149 0.600 0.600 0.600 0.600 Mass recovered (ugF-) 0,0635 0.131 0.113 0.188 0.342 0.360 0.316 0.344 0.306 0.258 0.142 0.0585 0.0691 0.0606 0.0496 0.0459 0.0371 0.0590 0.0414 0.0377 0.0827 0,0487 0.157 0.252 0.0668 0.0580 0.0478 0.0827 0.107 0.113 0.254 0.445 0.345 0.529 Sample ID SPIKE 250-5 blank blank F52809 DAY 4 F52724 DAY 4 F52737 DAY 4 F52801 DAY 4 F52720 DAY 4 F52733 DAY 4 F52788 DAY 4 F52730 DAY 4 F52731 DAY 4 F52802 DAY4 SPIKE 62-1 SPIKE 62-2 SPIKE 250-1 SPIKE 250-2 SPIKE 250-3 SPIKE 250-4 SPIKE 250 -5 SPIKE 250-6 SPIKE 250-7 SPIKE 250-8 SPIKE 124-1 BLANK BLANK BLANK BLANK BLANK BLANK F52714 DAY 4 F52734 DAY4 F52774 DAY 4 F52712 DAY 4 Actual Sample TISAB mL FC95 reading Qty rinavlol spiked (ppm F-) (mL org) (mL) 0.230 0.1 2.0 0.004 0.177 0.1 2.0 0.0331 0.1 2.0 0.0196 0.1 2.0 0.0187 0.1 2.0 0.0164 0.1 2.0 0.0359 0.1 2.0 0.0265 0.1 2.0 0.0218 0.1 2.0 0.0192 0.1 2.0 0.0174 0.1 0.0149 0.1 0.0232 0.1 0.0325 0.1 0.0722 0.1 0.130 0.1 0.123 0.1 0.199 0.1 0.0964 0.1 0.119 0.1 0.183 0.1 0.138 0.1 0.192 0.1 0.114 0.1 0.0825 0.1 0.0818 0.1 2.0 2.0 2.0 2.0 0.004 2.0 0.004 2.0 0.004 2.0 0.004 2.0 0.004 2.0 0.004 2.0 0.004 2.0 0.004 2.0 0.004 2.0 0.004 2.0 0.004 2.0 2.0 0.0578 0.1 2.0 0.0562 0.1 2.0 0.0495 0.1 2.0 0.0379 0.1 2.0 0.0290 0.1 2.0 0.0446 0.1 2.0 0.0285 0.1 2.0 0.0218 0.1 2.0 Cone..FC95 % Actual solution recovery ppm F- (ppm) (ug/ug) ir!Sample 250 77% 4.60 3.54 0.663 0.391 0.373 0.328 0.718 0.531 0.436 0.383 0.348 0.298 0.464 62 44% 0.649 62 97% 1.44 250 43% 2.59 250 41% 2.47 250 66% 3.98 250 32% 1.93 250 40% 2.38 250 61% 3.66 250 46% 2.76 250 64% 3.83 124 76% 2.27 1.65 1.64 1.16 1.12 0.990 0.758 0.580 0.892 0.570 0.436 Mass spiked (ugF-) 0.600 0.149 0.149 0.600 0.600 0.600 0.600 0.600 0.600 0.600 0.600 0.298 Mass recovered lugF-)_ 0.460 0.354 0.0663 0.0391 0.0373 0.0328 0.0718 0.0531 0.0436 0.0383 0.0348 0.0298 0.0464 0.0649 0.144 0.259 0.247 0.398 0.193 0.238 0.366 0.276 0.383 0.227 0.165 0.164 0.116 0.112 0.0990 0.0758 0.0580 0.0892 0.0570 0.0436 Sample ID F52729 DAY 4 F52803 DAY 4 F52716 DAY4 F52728 DAY 4 F52787 DAY 4 SPIKE 62-1 SPIKE 62-2 SPIKE 62-3 SPIKE 124-1 SPIKE 124-2 SERUMBLANK SERUM BLANK SERUM BLANK SERUM BLANK SERUM BLANK SPIKE 62-1 SPIKE 62-2 SPIKE 62-3 SPIKE 124-1 SPIKE 124-2* SPIKE 124-2* SPIKE 124-4 blank blank F52717 DAY4 F52735 DAY4 F52736 DAY4 F52726 DAY4 F52732 DAY4 F52739 DAY4 F52711 DAY 8 F5271 9 DAY 8 SPIKE 62-1 SPIKE 62-2 Actual Sample TISAB mL FC95 reading Oly finavlol spiked (ppm F-)(mL org) (mL) 0.0226 0.1 0.0237 0.1 0.0206 0.1 0.0191 0.1 0.0222 0.1 0.0307 0.1 0.0409 0.1 0.0710 0.1 0,0571 0.1 0.109 0.1 0.0397 0.1 0.0379 0.1 0,0379 0.1 0.0306 0.1 0.0277 0.1 0.0898 0.1 0.0913 0.1 0.0769 0.1 0.113 0.1 0.0326 0.1 0.270 0.1 0.121 0.1 0.0485 0.1 0.0299 0.1 0.0493 0.1 0.0319 0.1 0.0251 0.1 0.0256 0.1 0.0213 0.1 0.0164 0.1 0.0242 0.1 0.0164 0.1 0.0332 0.1 0.0500 0.1 2.0 2.0 2.0 2.0 2.0 2.0 0.004 2.0 0.004 2.0 0.004 2.0 0.004 2.0 0.004 2.0 2.0 2.0 2.0 2.0 2.0 0.004 2.0 0.004 2.0 0.004 2.0 0.004 2.0 2.0 2.0 0.004 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 0.004 2.0 0.004 Cone.FC95 solution (ppm) 62 62 62 124 124 62 62 62 124 124 62 62 % Actual recovery ppm F- _(ug/ug) insample 41% 55% 95% 38% 73% 121% 123% 103% 76% 81% 45% 67% 0.452 0.474 0.411 0.381 0.443 0.615 0.817 1.42 1.14 2.19 0.794 0.758 0.758 0.612 0.554 1.80 1.83 1.54 2.25 0.652 5.40 2.41 0.970 0.597 0.987 0.638 0.503 0.511 0.425 0.328 0.484 0.328 0.663 0.999 Mass spiked (ugF-) 0.149 0.149 0.149 0.298 0.298 0.149 0.149 0.149 0.298 0.298 0.149 0.149 Mass recovered (ugF-) 0.0457' 0.0474 0.0411 0.0381 0.0443 0.0615 0.0817 0.142 0.114 0.219 0.0794 0.0758 0.0758 0.0612 0.0554 0.180 0.183 0.154 0.225 0.0652 0.540 0.241 0.0970 0.0597 0.0987 0.0638 0.0503 0.0511 0.0425 0.0328 0.0484 0.0328 0.0663 0.100 STLAIR@ Sample ID SPIKE 62-3 SPIKE 62-4 SPIKE 62-5 SPIKE 62-6 SPIKE 62-7 SPIKE 62-8 SPIKE 62-9 SPIKE 124-1 SPIKE 124-2 BLANK BLANK F52725 DAY 8 F52721 DAY 8 F52727 DAY 8 F52806 DAY 8 F52713 DAY 8 F5271 8 DAY 8 F52723 DAY 8 F52715 DAY 8 F52738 DAY 8 F52809 DAY 8 SPIKE 62-1 SPIKE 62-2 SPIKE 62-3 SPIKE 62-4 SPIKE 124-1 SPIKE 124-2 SPIKE BLANK BLANK SERUM SERUM SERUM SPIKE 124-3 BLANK BLANK BLANK 62-1 Actual Sample reading Oty (ppm F-) (mL or 0.0498 0.1 0.0392 0.1 0.126 0.1 0.0354 0.1 0.0421 0.1 0.0508 0.1 0.0537 0.1 0.0710 0.1 0.144 0.1 0,0847 0.1 0.0392 0.1 0.0300 0.1 0.0278 0.1 0.0230 0.1 0.0231 0.1 0.0170 0.1 0.0175 0.1 0.0173 0.1 0.0172 0.1 0.0165 0.1 0.0152 0.1 0.0245 0.1 0.0353 0.1 0.0414 0.1 0.0579 0.1 0.0925 0.1 0.0865 0.1 0.1048 0.1 0.0868 0.1 0.0414 0.1 0.0310 0.1 0.0284 0.1 0.0195 0.1 0.0558 0.1 TISAB mL FC95 finalvol spiked (mL)__ __ 2.0 0.004 2.0 0.004 2.0 0.004 2.0 0.004 2.0 0.004 2.0 0.004 2.0 0.004 2.0 0.004 2.0 0.004 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 0.004 2.0 0.004 2.0 0.004 2.0 0.004 2.0 0.004 2.0 0.004 2.0 0,004 2.0 2.0 2.0 2.0 2.0 2.0 0.004 Conc.FC95 solution (ppm) 62 62 62 62 62 62 62 124 124 62 62 62 62 124 124 124 62 % Actual recovery ppm F- (ug/ug) Insample 67% 0.995 53% 0.784 169% 2.52 48% 0.709 57% 0.842 68% 1.02 72% 1.07 48% 1.42 96% 2.87 1.69 0.784 0.600 0.556 0.461 0.462 0.341 0.349 0.346 0.344 0.331 0.303 33% 0.490 47% 0.706 56% 0.828 78% 1.16 62% 1.85 58% 1.73 70% 2.10 1.74 0.828 0.620 0.568 0.390 75% 1.12 Mass spiked (ugF-) 0.149 0.149 0.149 0.149 0.149 0.149 0.149 0.298 0.298 0.149 0.149 0.149 0.149 0.298 0.298 0.298 0.149 Mass recoverei lugF-) 0.100 0.0784 0.252 0.0709 0.0842 0.102 0.107 0.142 0.287 0.169 0.0784 0.0600 0.0556 0.0461 0.0462 0.0341 0.0349 0.0346 0.0344 0.0331 0.0303 0.0490 0.0706 0.082E 0.116 0.185 0.173 0.210 0.174 0.082 0.062 0.056 0.039 0.112 Sample ID SPIKE 62-2 SPIKE 62-3 SPIKE 124-1 SPIKE 124-2 SPIKE 124-3 SPIKE 124-4 BLANK BLANK BLANK F52724 DAYS F52737 DAYS F52801 DAYS F52720 DAYS F52733 DAYS F52788 DAYS spike62-1 spike62-2 spike 124-1 Actual Sample TISAB mL FC95 reading aty finalvol spiked (ppm F-) (mL org) (mL) 0.06ig 0.1 0.0604 0.1 0.0933 0.1 0.0551 0.1 0.109 0.1 0.109 0.1 0.0837 0.1 0.0367 0.1 0.0305 0.1 0,0289 0.1 0.0282 0.1 0.0428 0.1 0.0394 0.1 0.0245 0.1 0.0129 0.1 0.0430 0.1 0.0518 0.1 0.104 0.1 2.0 0.004 2.0 0.004 2.0 0.004 2.0 0.004 2.0 0.004 2.0 0.004 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 0.004 2.0 0.004 2.0 0.004 Conc.FC95 solution (ppm) 62 62 124 124 124 124 62 62 124 % Actual recovery ppm F- (uglug) insample 83% 1.24 81% 1.21 63% 1.87 37% 1.10 73% 2.19 73% 2.17 1.67 0.734 0.610 0.578 0.563 0.856 0.788 0.491 0.259 58% 0.860 70% 1.04 70% 2.08 Mass spiked (ugF-) 0.149 0.149 0.298 0.298 0.298 0.298 0.149 0.149 0.298 Mass recovered (ugF-) 0.124 0.121 0.187 0.110 0.219 0.217 0.167 0.0734 0.0610 0.0578 0.0563 0.0856 0.0788 0.0491 0.0259 0.0860 0.104 0.208 Forgotto spikeblankserum withFC-95 thereforea,notherreplicatweas analyzed. DP2 (08/17/95) Instrumentmalfunctiont,he boatdid notgo allthe way Intothe oven, anotherreplicatweas analyzed. DP2 (08/21/95 Samples reanalyzedon followindgay, due to QC not meeting requirements.DP2 (08/21/95)