Document wkag7JO1EQQGa2xqa7rOb66
/H U & -0 3I
OCTANOUWATER PARTITION COEFFICIENT
TEST SUBSTANCE
Identity: N[r[e(2fhe-e(rNpret-admdteeotchaaysfllpuNeo-rrMfoluoeocFrtOoyolS)csEtuaAlfn,oenosyrulM]lmfoeenFtahOmySliadEmoA)i.-neo(th]2ey-tPlhrayoclpreyeslnateoteri,c; CamcAaiSdy,#a2l-so be 25268-77-3)
crReoemmmapaionruiknnsgd: 5a%sM"oTaf-tt5eh8rei6ap9l r"iso(dBaunactcat hmwa2bs4e0rn,8ow)t.acxShyaarsmaocplitldee.ripzTuehrdiet.yteissgt ilvaebnoraasto"r9y5i%de"n. tiTfihees the
METHOD
AYMGenLeaaPthrlyo(cYtdoic/:mNa)lp:: letYOHeePEdsLC: CD
117 & EEC-Directive 92/69 EEC, w19ith94Spectrophotometric detector
Annex
V,
Part
A.8
RrloegefemPreoawnr-ckess: . TTheessteinsguubssetadnsciexsswubesretanusceeds wtoitchaklinborawten tlhoeg ePlouwtiovanlutiemseaisn units of
Reference substance Benzylalcohol Toluene 1,4-dichlorobenzene 2FD,li4ub-oeDrnaDznyTtlh((e99n99e%%())98%)
log1.1Pow 2.7 4463....8274
A stock solution of the test substance was prepared by dissolving 287 mg in 50.0 tmiml_eascwetiothnimtriolebialendphsaosneic,arteinsgultfionrg2inmthineutteess.t sTohluetisotnoc(k57s4olmutgio/Ln).was diluted 10
Mreospbeilectpivhealys.e is a 75/25 (v/v) solution of acetonitrile / Millipore-Q water
RESULTS
Substance Reference chemicals
TB1o,e4lnu-dzeyinclaehllcoorohboel nzene
log Pow
o| ^ **
23..74****
000095
2FD,li4ub-oeDrnaDznyTtlhene Test substance
SPPPeeeevaaaekkkr23a1l(smmaajollrpceoamkps.)
446...782****** 5.6* -0.557../435**.2*
Interpolated from the regression line:y = 0.288x - 0.730 (r=0.972, n=6)
** log Pow values according tothe OE C D guidelines.
CONCLUSIONS
This study estimates the log Powfor MeFOSEA.
Submitter: 3M Company, Environmental Laboratory, P.O. Box 33331, St. Paul, Minnesota, 55133
DATA QUALITY
Reliability: Klimisch ranking 2
-per OECD. HPLC methods enables partition coefficients to be estimated in the log Pow range between 0 and 6.
-Notox states impurities are of minor importance in HPLC method, however OECD states, HPLC method is less sensitive to the presence of impurities in the test substance than the Shake Flask method. Nevertheless, in some cases pimepaukraitisessigcnamnemntask.eMthixetuinretesrpwrheitcahtiornesoufltthine arensuunltrsedsoiflfviceudltbdaunedt,ouupnpceerratanidnty in lower limits of log Powshould be stated. Duplicate analysis should be reported in orardngeer toofs+h/-ow0.1colongfiduennitcse) -indthaetampreoavsidueredmmeenat n(lovgalPueowthvaelrueefosreshcoaunld't fsahllowwithin a confidence. pH of eluant is critical of ionizable compounds. The study did not include any fluorochemicals among the reference substances. Without fluorochemical reference compounds, it is not known whether the same relationship between the capacity factor k' and Kow exists for compounds with perfluoroalkyl portions as exits for nonfluorinated reference materials such as those used in this study. The reference compounds should have been also irnejfeecrteendceincdoempepnoduenndt.lyRtoefeinrseunrceeaspupbrsotparniacteesrweteernetioinnjetcimteed atwssicigen, ehdowtoeveearchonly the mean retention time data is given for the reference substances thus repeatability of the retention times is not clear. It was not demonstrated that the test material was soluble in the acetonitrile stock solution or in the mobile phase. No demonstration that the major components of the test substance are dmeatienctcaobmlepoant eUnVtsXo=f t2h1e0tensmt.sUubVstaadnscoer.pAtiosnsusmpepctitoransisanreeemdaeddeotfhtahtethpeurmifieadjor
000096 -
wpeaaskmfraodme tthoecHonPfLirCm othfitshe(et.egs. tMsausbsstsapnecce aisnathlyesmisatojosrhcoowmpthoenemnat.teNrioaladtteetmecptted choams teheofaf tphperocporliuamtenm. aSsosm).eItkis' vnaoluteksnorwepnowrtehdetahreer cthaelcmulaatjeodr cionmcoprroencetlnyt.s even REFERENCES SEtnuvdiryoncomnednutcatleRdeastetahrechreBq.uVe.,stTohfe3NMeCthoemrlapnadnsy, bAyprNilO7T, O19X9S4.afety & OTHER Last changed: 5/17/00
000097
REPORT
DETERMINATION OF THE PARTITION COEFFICIENT (N-OCTANOL/WATER) OF T-5869 i L\
BY HIGH PERFORMANCE LIQUID CHROMATOGRAPHY (HPLC)
NOTOX Project 121286 NOTOX substance 38196
- Page 1 of 14 -
000038
RECEIVED
NOV - 9 1994
TOXIC3OM LOGY
T-5869
NOTOX Project 121286
STATEMENT OF GLP COMPLIANCE
NOTOX B.V., 's-Hertogenbosch, The Netherlands
The study described in this report was conducted in compliance with the most recent edition of: The OECD Principles of Good Laboratory Practice which are essentially in conformity with: The United States Food and Drug Administration. Title 21 Code of Federal Regulations Part 58. The United States Environmental Protection Agency (FIFRA). Title 40 Code of Federal Regulations Part 160. The United States Environmental Protection Agency (TSCA). Title 40 Code of Federal Regulations Part 792.
Study Director:
Drs. R. de Vries
Date:
-SV
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NOTOX Project 121286
QUALITY ASSURANCE STATEMENT NOTOX B.V., 's-Hertogenbosch, The Netherlands.
Study procedures were subject to periodic inspections and general non study specific processes were also inspected at periodic intervals.
This report was audited by the NOTOX Quality Assurance Unit and the methods and results accurately reflect the raw data.
DATES OF QAU INSPECTIONS/ AUDITS
REPORTING DATES
25 March 1994 28 March 1994 21 April 1994
25 March 1994 28 March 1994 21 April 1994
Quality Assurance Manager
C.J. Mitchell B.Sc.
Date: 0 e!- Ob'St, .
- Page 3 -
000100
T-5869 REPORT APPROVAL STUOY DIRECTOR: <* MANAGEMENT:
O
NOTOX Project 121286
Ors. R. de Vries
O.A.M.W. van Helvoirt Section head Physico Chemistry
,r\
PPJ
Dr.^lona C. Enninga [Technical Director
Date: O 0 / o b i * 0 0 4 1
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L
T-5869
NOTOX Project 121286
PREFACE Sponsor
Study Monitor Testing Facility
Study Director Study plan
TEST SUBSTANCE
Identification Description Batch Purity Storage conditions Stability under storage conditions Expiry date
3M Belgium - Chemical EBC Canadastraat 11 B-2070 ZWIJNDRECHT Belgium Mr. R.H. Cox NOTOX B.V. Hambakenwetering 3 5231 DD 's-Hertogenbosch The Netherlands Ors. R. de Vries Start: 30 March 1994 Completed: 07 April 1994
T-5869 Cream solid 2408 95% At room temperature in the dark
Stable January 01, 1996
l.
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T-5869
NOTOX Project 121286
PURPOSE AND PRINCIPLE
The purpose of the study was to determine the partition coefficient of the test substance between n-octanol and water.
The partition coefficient (n-octanol/water) (Pow) is defined as the ratio of the equilibrium concentrations in a two phase system consisting of n-octanol and water.
The retention time of a substance in a reversed-phase High Performance liquid Chromatography (HPLC) chromatographic system is related to its partition coefficient (n-octanol/water). The HPLC method is useful for log Pow values between 0 and 6. Using the HPLC method, impurities are of minor importance. The HPLC method must also be applied if the test substance is a mixture of compounds but is not applicable to strong acids and bases, metal complexes, substances which react with the eluent or surface-active agents.
GUIDELINES
The study procedure described in this report was based on the following guidelines:
Organization for Economic Co-operation and Development (OECD), OECD Guideline for Testing of Chemicals, guideline no. 117: "Partition coefficient (n-octanol/water) High Performance Liquid Chromatography (HPLC) method" (adopted March 30, 1989).
European Economic Community (EEC), EEC-Directive 92/69 EEC, Annex V, Part A, Methods for the determination of physico-chemical properties, A.8: "Partition coefficient". EEC Publication no. L383, December 1992.
ARCHIVING
NOTOX B.V. will archive the following data for at least 10 years: protocol, report, test substance reference sample and raw data.
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TEST SYSTEM AND RATIONALE
A High Performance Liquid Chromatograph with a spectrophotometric detector. The stationary phase was bonded silica C^g. The mobile phase was 75/25 (v/v) acetonitrile/water. Both HPLC instrumentation and conditions are described in the section "method of chemical analysis".
The test system was recognized by the international guidelines (OECD, EEC).
REFERENCE SUBSTANCES
Six chemicals for which log Pow has been reported were used to calibrate the elution time in units of log Pow. The mixture of reference substances
consisted of:
Reference substance
- benzylalcohol (p.a., Merck) - toluene (p.a., Merck) - 1,4-dichlorobenzene (z.s., Merck) - fluoranthene (98%, GC, Aldrich Chemie) - dibenzyl (99%, GC, Aldrich Chemie) - 2,4-DDT (99%, HPLC, Riedel de Haen, Seelze, FRG)
1 log PQW values according to the OECD guideline.
log Pqv/
1.1 2.7 3.4 4.7 4.8 6.2
PERFORMANCE OF THE TEST
Preparation of the solutions
Solutions of the reference substances were prepared in methanol (HPLC-grade, Labscan Limited Co., Dublin, Ireland) at a concentration of 1.5-2 g/1. A mixture of the reference substances was prepared by adding 125 pi of each solution to a volumetric flask. Subsequently, this flask was made up with mobile phase to a final volume of 25.0 ml. For the determination of t0 (retention time of the unretarded component), a solution of formamide (p.a., Merck, Darmstadt, FRG) in methanol was prepared at a concentration of 1.7 g/1 and thereafter diluted 10 times with mobile phase. A stock solution of T-5869 was prepared by dissolving 287 mg test substance in 50.0 ml acetonitrile (2 minutes sonication). The stock solution was diluted 10 times with mobile phase, resulting in the test solution.
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NOTOX Project 121286
Performance of the test
The solutions were injected in the following sequence: the mixture of reference substances, mobile phase, the formamide solution, the test solution (in duplicate), mobile phase, the mixture of reference substances and the formamide solution.
Temperature of measurement
The temperature of the mobile phase was recorded several times during the measurements.
METHOD OF CHEMICAL ANALYSIS
The conditions used for the High Performance Liquid Chromatographic method are described below:
Analysis
Column
Mobile phase
Flow Detection Injection volume
LiChrospher 100 RP-18; 125 x 4 (I.D.) mm;
dg= 5 ym (Merck, Darmstadt, FRG) 75/25 (v/v) acetonitrile (HPLC-grade, Labscan Limited Co., Dublin, Ireland)/Milli-Q water (Mi 11 ipore Corp., Bedford, MA, USA) 1 ml/min UV at X= 210 nm 10 yl
Instrumentation
HPLC Pump Autosampler Detector
Integrator
Series 410 LC (Perkin Elmer, Norwalk, CT, USA) ISS-200 (Perkin Elmer) SpectroMonitor 3100 (LDC Analytical, Riviera Beach, FL, USA) SP 4290 (Spectra Physics, San Jose, CA, USA)
DATA HANDLING
Using High Performance Liquid Chromatography, especially large log Pow values can be accurately determined^rom the chromatographic retention data To this end, the capacity factor (k') was used, since it is proportional to the partition coefficient. The capacity factor was calculated from the retention of the substance concerned (tr) and the unretarded component (t0)
k = (tp - tQ)/t0
From the results of the reference substances, a plot of log Pow (x-value)
versus log k ' (y-value) was constructed, using linear regression analysis. The log k^ value of each component of the test substance was compared with the log k ' values of the reference substances with known log Pow values.
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RESULTS
Representative HPLC chromatograms of the test solution, the blank (mobile phase), the mixture of reference substances and the formamide solution are shown in Figures 1 to 4.
In the HPLC chromatograms of T-5869, using detection at 210 nm, one large and 2 small test substance peaks were observed. In addition, many small test substance peaks were observed between approximately 1 and 6 minutes. It was assumed that the large peak derived from the major component of test substance whereas the small peaks derived from impurities.
The t0 was determined to be 0.90 minutes as a mean value of both measurements (i.e 0.89 and 0.90 minutes).
The mean values of the retention times, k' values, log k ' values, log PgW and Pow values^are summarized in Table 1. The plot of log Pow (x-value) versus log k ' (y-value) from the reference substances is shown in Figure 5.
Table 1 Results of the test.
Substance
trx k '
log k' log Pow
pow
benzyl alcohol toluene 1,4-di chlorobenzene fluoranthene dibenzyl 2,4-DDT
test substance peak 1 (major comp.) peak 2 peak 3 several small peaks
1.14 2.24 2.89 5.67 4.07 9.12
7.75 6.96 6.60 1-6
0.274 1.503 2.229 5.330 3.547
9.190
-0.563 0.177 0.348 0.727 0.550 0.963
1.1 2.7 3.4 4.7 4.8
6.2
7.659
0.884
5.6 2
3.98x10s
6.777
0.831
5.4 2
2.51x10s
6.374
0.804
5.3 2
2.00x10s
0.12/ 5.67 -0.93/0.75 -0.7/5.2 2 0.2/1.58x10s
1 Mean value of the retention times of both chromatograms.
1 Interpolated from the regression line: y = 0.288x - 0.730 (r=0.972, n=6) Note : The calculations were performed using not rounded values.
The temperature of the mobile phase was 20.2-20.6C during the measurements.
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NOTOX Project 121286
HATA SAVED TO BIN # 50
38196/1212 86
07-04-94 08:03:!
,f*>
ILE 1.
METHOD 0.
RUN 7
INDEX
PERK#
RREfiX
RT AREA ec
1 2 3
0. 041 0. 209 0. 098
0. 15 0. 25 0. 34
1321 82 6658 02 3122 02
4
5
"617
8
9
10
11
12
13
14
15
IS
17
18
19
20
21
22
2 2
4/
3. 425 0. 121 0. 251 0. 498 0.189 8. 25 9. 428 8. 839 0. 899 3. 097 0. 092 1. 652 1. 125 4. 167 8. 248 0. 472 1. 217 0. 89 13. 076 11. 364 55. 39
0- 54
8. 7-9
1. 02
1. 1
1. 29
1. 47
1. 58
1. 72
2. 03
2. 48
2. 93
3. 15
7. 9
4.'13
4. 83
5, 23
5. 59
5. 88
66.. 7
9671 77
189239 08
3874 05
8011 06
15893 06
6033 06
7978 06
1.3648 06
26774 06
28669 07
98772 88
2936 06
52708 03
35886 02
132925 03
7895 02
13795 02
38811 02
28393 02
43 6127409843 17667 96
02 02 07
7
PS= 1. BIN 50
Figure 1 HPLC chromatogram of the test solution of 574 mg/1 T-5869 in mobile phase.
i.
Column
LiChrospher 100 RP-18; 125x4 (I.D.) mm; dp=5 pm
Mobile phase
75/25 (v/v) acetonitrile/Milli-Q Water
Flow
1 ml/min
Detection
UV, at X = 210 nm
Injection volume 10 yl
Note: All the peaks between 0 and approximately 1 minute and some of the peaks between approximately 1 and 2 minutes were also observed in the chromatograms of the blank (see Figure 2). Therefore it was assumed that these peaks did not derive from the test substance.
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CHANNEL A
INJECT 07-04-94 9:74:04 STORED TO BIN # 56
DATA SfiVED TO BIN # 56
38196/121286
FILE 1.
METHOD
ANALYST : KRH
AK#
AREA/.
1 1. 543
cL 10.515
3. 853
4 55. 335
cr J
7. 76s
f. 6. 901
2. 509
8 11.576
OTAL
100.
7-04-94 9 : <4:04 0. RUN 13 INDEX 1
RT
0. 15 0. 25 0. 34 0. 52 0.67 0. 99 1. 18 1-61
AREA ec
877 02 5976 02 21 90 02 31449 02 4415 03 3922 02 1426 03 6579 01
56834
CH= "A" PS= 1. BIN 56
Figure 2
HPLC chromatogram of a blank (mobile phase).
Column
Mobile phase Flow Detection
Injection volume
LiChrospher 100 RP-18; 125x4 (I.D.) mm; dp=5 ym
75/25 (v/v) acetonitrile/Milli-Q water 1 ml/min UV, at X = 210 nm 10 yl
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CHANNEL fi
INJECT 07-04-94 09:50:17 ST OR EU TO BIN # 57
ER 0 DATS SAVED TO BIN # 57
38136/12 1286
FILE 1.
METHOD
ANALYST KRH
PERK#
RRERX
1 0 823
Cl 0. 145
s 0. 06
4
c
8. 873 0. 873
6 0. 133
7 8. 977
8 0. 112
3 11.317
10 0. 013
11 10. 567
12 15.524
13 38. 584
14 0. 316
15 13. 272
TOTAL
100.
07-04- 34 83:50:17 0. RUN 14 INDEX 14
RT fiRFfi BC
0. 14 8. 25 0. 34 0. 53 0. 73 0, 96
I. 14
1'". 6".1*A+ 2. 61 2. 83 4. 08 5. 68 6. 98 9. 13
-1131 R2 7039 02 29-1 7 02 42751 88 3542 @5 6771 06 4 <6774 03 5460 01 558640 01
611 02 514173 03 755334 01 1877339 01
15361 01 645770 01
4865613
CH= "fi" PS= 1. BIN 57
Figure 3 HPLC chromatogram of the mixture of reference substances in mobile phase.
Column Mobile phase Flow Detection Injection volume
LiChrospher 100 RP-18; 125x4 (I.D.) mm; dD=5 pm
75/25 (v/v) acetoni tri le/Mi 11i-Q water H 1 ml/min
UV, at X = 210 nm 10 pi
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CHANNEL ft
INJECT 0 7 - 0 4 - 9 4 7 :77:44 STORED TO BIN # 49
DfiTfi
38196/121286
FILE 1.
METHOD
fiNfiLVS:Ts KRH
PEftK#
ftREfti
1 9. 35
2 1. 189
3 0. 79
4 10. 44
5 1. 884
6 54.548
1* 6.246
y 5
0.12 3, 094
10 0. 424
1 1 20. 915
TOT fL
100.
07-04- 94 0 7 :7 7 : 4 4 0. RUN 6 INDEX 6
RT
0.16 0.27
J
.55 0. 78 0.89 1 -1 1 1. 42 1 .87 2. 97
S,
BRER BC
2624 02
8926 82
5930 02
78363 08
14143 06
40Q4>3779
88 05
899 05
23224 92
3185 03
1.56989 01
750599
CH= "fiM PS= 1. BIN 49
Figure 4 HPLC chromatogram of a formamide solution of 0.2 g/1 in mobile phase.
Column
Mobile phase Flow
Detection Injection volume
LiChrospher 100 RP-18; 125x4 (I.D.) mm; dD=5 ym 75/25 (v/v) acetonitrile/Milli-Q water H 1 ml/min
UV, at X = 210 nm 10 yl
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Figure 5 Plot of log Pow (x-value) versus log k ' (y-value) from the reference substances.
