Document LJeBOk9pqGgadvR3pmwddYwRg

TECHNICAL REPORT SUMMARY TO: TECHNICAL COMMUNICATIONS CENTER - 201-2CN BACK TO MAIN IDiltv 8/25/81 1 - (1ttitwrt;iiit I f report is-prititcd on tmth sides of paper, send two copies to TCC.) Division -E..n-v i r o n m e n t a l L a b o r a t o r y (EE & PC) I'rojtn.~ IIDspt. Number 0535 1Proior.t Ntrmliur Commercial Chemicals Division Report Title B i o s o r p t i o n of CS-2151 (CS-2070) TO 3 I[Re9p9or7t N0um0b1e2r 600 D. R . l i i c k e r , Commercial C h e m i c a l s D i v i s i o n , 53-4N Aulhor(sJ E. A . R e i n e r , E n v i r o n m e n t a l L a b o r a t o r y / E E & PC, 21-2W-05 I E m plo ee Num 4$816 b e r ( s 1 NOIcljook Rvforortco No. of Pagos Including Coversheel None (see L a b Request 6457 S) SECUR'TY I3 Open (Company Confidential) 0 Closed (Special Authorization) 3M CHEMICAL RElGlSTRY New Chemicals Reported 0 yes E4 No KEYWORDS: (Select terms from 3M Thesaurus. Suggest other applicable terms. 1 CUR RENT OBJECTIVE:- To measure t h e b i o s o r p t i o n p a r t i t i o n c o e f f i c i e n t of CS-2070. EE & PC D I V (ENV. LAB) Bisorption Bioconcentration Microorganisms Fluorochemical Surfactants Waste T r e a t m e n t ~~ ~~ ~ REPORT ABSTRACT: (200-250words) This abstract information is distributed by the Technical Communications Canter to alert 3M'ers t o Company R&D. It i s Company confidential material. A 0.125 m g / l s o l u t i o n of CS-2070 ( t h g s o l i d s p o r t i o n of CS-2151) w a s s h a k e n f o r 1 h o u r a t 2!5 C i n t h e p r e s e n c e of 2,000 m g / l of v i a b l e o r hea.t k i l l e d a c t i v a t e d s l u d g e . A f t e r removing solids by c e n t r i f u g a - t i o n , t h e t e s t material c o n c e n t r a t i o n w a s determined as the! d i f f e r e n c e between t h e T O F * c o n c e n t r a t i o n of t h e t e s t and c o n t r o l c u l t u r e s . The results showed t h a t m o s t of t h e CS-2070 shaken i n t h e p r e s e n c e of v i a b l e a c t i v a t e d sludge! remained i n t h e s o l u t i o n . On t h e o t h e r h a n d , h e a t k i l l e d s l u d g e a p p a r e n t l y adsorbed CS-2070. *TOF is T o t a l O r g a n i c F l u o r i n e . Information Liaison Init i d s : BACK TO MAIN BIOSORPTION OF CS-2151 (CS-2070) I ntrod uct i o n : This report describes tests p e r f o r m e d o n CS-21511 t o e v a l u a t e i t s b i o s o r p t i o n to t h e v o l a t i l e p o r t i o n of a c t i v a t e d sludge mixed liquor. Since CS-2151 is not l i k e l y to biodegrade, this; t e s t i n q i n d i c a t e s w h e t h e r t h e s u b j e c t material is l i k e l y t o be removed i n waste treatment through sorption to sludge. This testing is also useful i n p r e d i c t i n g whether CS-2151 is l i k e l y t o accumulate t h r o u g h s o r p t i o n t o t h e o r g a n i c p o r t i o n of s o i l a n d aquatic s e d i m e n t s . Methods and Materials: ' I . L I ~t 2 1 ~ d . L L u i i : i t e r i c d lLaLJoriitxry received t h e t e s t sample o n 1 1 / 4 / 8 0 . The s a m p l e was a l i g h t y e l l o w l i q u i d . I t was r e p o r t e d t o c o n t a i n 258 s o l i d s i n water. The s o l i d p o r t i o n of t h i s product is d e s i g n a t e d CS-2070. Greg Vraspir p e r f o f i e d t h i s work i n accordance w i t h t h e a t t a c h e d E n v i r o n m e n t a l L a b o r a t o r y Protocol. L. D. W i n t e r of t h e Commercial Chemicals LabQratory performed t h e a n a l y t i c a l work. S i n c e a s u f f i c i e n t l y sensitive specific a n a l y t i c a l technique was not a v a i l a b l e , t h e f i n a l CS-2070 c o n c e n t r a t : i o n was d e t e r m i n e d by m e a s u r i n g t h e d i f f e r e n c e between t h e t o t a l o r g a n i c f l u o r i n e (TOF) c o n c e n t r a t i o n i n t h e CS-2070 c o n t a i n i n g sample and i d e n t i c a l l y t r e a t e d c o n t r o l samples n o t c o n t a i n i n g CS-2070. Results and Discussion: R e s u l t s of t h i s work are shown i n T a b l e 1 below: T a b l e I. n i o s o r p t i o n of 0.125 m g / l of CS-2070(1) o n 2 , 0 0 0 m g / l of heat k i l l e d and v i a b l e a c t i v a t e d sludge, Condition of Sludge .TOF(2) Expressed as T e mg/l st CS-2070 Control - F i n a l CS-2070 Concentration (mg/l) (by difference) H e at K i 1l e d 1.1 1.1 0 Viable 097 .84 013 Sludge Free So C rption ontrol ( 3 ) .15 (002 .15 .10 Footnotes: (1) CS-2151 i s a 25% a q u e o u s s o l u t i o n o f CS-20'70. ( 2 ) TOF is t o t a l o r g a n i c f l u o r i n e . ( 3 ) The sorption control is a CS-2070 s o l u t i o n t h a t was n e i t h e r centrifuged nor exposed to t h e plastic centrifuge tubes. BACK TO MAIN Biosorpt i o n of CS-2151/EAR Page 2 August 25, 1981 A f t e r e x p o s u r e t o h e a t k i l l e d s l u d g e t h e TOF c o n c e n t r a t i o n s o f t h e t e s t a n d c o n t r o l c e n t r i f u g a t e s were i d e n t i c a l . T h i s s u g g e s t s t h a t CS-2070 is a b s o r b e d by h e a t k i l l e d o r g a n i s m s . V i a b l e o r g a n i s m s o n t h e o t h e r hand, d i d n o t a b s o r b CS-2070. T h i s d i f f e r e n c e between t h e s o r p t i o n of t h e t e s t material by h e a t k i l l e d and v i a b l e c e l l s was n o t t o t a l l y unexpected. I n a l l t h e b i o a d s o r p t i o n s t u d i e s r e p o r t e d by S m i t h e t a l . (1, 2 ) i n which t h e y m e a s u r e d t h e b i o s o r p t i o n o f hydrocarbons by both h e a t k i l l e d and viable cell c u l t u r a l s they found h i g h e r s o r p t i o n c o e f f i c i e n t s w i t h h e a t k i l l e d c e l l s . The reason f o r t h i s d i f f e r e n c e is n o t known. Two a d d i t i o n a l p a r a l l e l c o n t r o l s were :run i n t h i s e x p e r i m e n t , a s l u d g e f r e e c o n t r o l and a c o n t r o l t o a i d i n deterrniniing w h e t h e r CS-2070 sorbed to t h e plastic c e n t r i f u g e tubes. These controls measured t h e CS-2070 c o n c e n t r a t i o n s i n t h e a b s e n c e of i n t e r E e r i n g o r g a n i c f l u o r i n e from t h e a c t i v a t e d s l u d g e , a n d t h u s showed t h e p r e c i s i o n of t h e a n a l y t i c a l t e c h n i q u e . The measured CS-2070 concentration i n t h e s l u d g e f r e e c o n t r o l w a s 0.025 m g / l greater thain t h a t i n i t i a l l y added, w h i l e t h e s o r p t i o n c o n t r o l c o n c e n t r a t i o n w a s 0.025 mg/l less. The p r e c i s i o n of t h e measurements is too l o w to allow t h e c a l c u l a t i o n o f b i o s o r p t i o n c o e f f i c i e n t s f o r CS-2070. However, t h e d a t a do i n d i c a t e t h a t CS-2070 is l i k e l y t o remain i n aqueous s o l u t i o n d u r i n g passage through a wastewater t r e a t m e n t system. They also s u g g e s t s t h a t CS-2070 i s n o t l i k e l y to a d s o r b to microorganisms or o t h e r o r g a n i c s i n the a q u a t i c or soil environments. References: 1 S m i t h e t a l , E n v i r o n m e n t a l Pathways; of S e l e c t e d C h e m i c a l s i n F r e s h w a t e r S y s t e m s , P a r t I , SRI I n t . e r n a t i o r r a 1 , EPA C o n t r a c t #68-3-2227, EPA-600/7-77-113, O c t o b e r , 197'7, NTIS tPB-274 548. 2 Smith e t al, E n v i r o n m e n t a l Pathwaysl of Selected C h e m i c a l s i n F r e s h w a t e r S y s t e m s , P a r t 11, EPA-600/7-78-074, May, 1978. BACK TO MAIN METHOD FOR MEASURING BIOSORPTION TO ACTIVATED SLUDlSE EAR 1/7/81 INTRODUCTION In t h e i r report on experimental procedures for studying environm e n t a l p a t h w a y s o f c h e m i c a l s i n f r e s h w a t e r s y s t e m s (5.1), S m i t h e t a1 s t a t e t h a t biosorption is an important parameter since it may a f f e c t b i o m a g n i f i c a t i o n up t h e f o o d c h a i n , and t h e a v a i l a b l e c o n c e n t r a t i o n f o r b i o d e g r a d a t i o n . The:y i n d i c a t e d t h a t t h i s phenomenon may a l s o a f f e c t t h e v i a b i l i t y of m i c r o o r g a n i s m s t h a t p a r t i c i p a t e i n t h e compounds biodegradation. S t i l l another reason f o r performing such tests is t h a t bios o r p t i o n may be a s i g n i f - i c a n t mode o f removal o f chemicals d u r i n g its passage through a wastewater treatinent system. The land d i s p o s a l o f t h e s l u d g e f r o m t h e s e treatment sylstems may b e a s i g n i f i c a n t mode of t r a n s p o r t i n g t h e chemical t o t e r r e s t r i a l e c o s y s t e m s . S u b s e q u e n t u p t a k e and a c c u m u l a t i o ~ nby p l a n t s may move t h e c h e m i c a l i n t o t h e human food c h a i n . T h i s p r o c e d u r e i s a simple method of m e a s u r i n g b i o s o r p t i o n . I t is modeled a f t e r t h a t o f Smith e t a1 (!5.1, 5.2) b u t d i f f e r s i n t h a t i t calculates' t h e b i o s o r p t i o n p a r t i t i o n c o e f f i c i e n t ( KP) based on t h e v o l a t i l e suspended s o l i d s (VSS) c o n c e n t r a t i o n i n a c t i v a t e d s l u d g e . The method o f Smith e t a1 u s e s a m i x t u r e o f l a b o r a t o r y - g r o w n p u r e c u l t u r e s . T h i s use o f a n a t u r a l l y d e v e l o p e d m i x e d p o p u l a t i o n may cause some v a r i a b i l i t y i n r e s u l t s due to v a r i a t i o n i n t h e population structure of d i f f e r e n t a c t i v a t e d s l u d g e samples, b u t t h i s v a r i a b i l i t y should be small. Smith e t a1 interpreted d a t a they generated (5.1) to indicate t h a t t h e v a l u e of Kp f o r a s p e c i f i c type of e n v i r o n m e n t a l s a m p l e s h o u l d n o t v a r y b y more t h a n a f a c t o r of 3 . Small v a r i a b i l i t y w i l l n o t a f f e c t t h e major p u r p o s e s o f t h e t e s t d e s c r i b e d above, t h u s t h e p r e c i s i o n requirements o f t h i s method d o n ' t j u s t i f y t h e m a i n t e n a n c e of p u r e c u l t u r e s i n t h e l a b o r a t o r y . The method assumes t h a t a l l s o r p t i o n is t o t h e v o l a t i l e p o r t i o n of t h e a c t i v a t e d sludge. The v a l i d i t y of t h i s assumption is s u b s t a n t i a t e d by t h e f a c t t h a t most s o r p t i o n t o s o i l c a n be a t t r i b u t e d t o i t s o r g a n i c c o n t e n t (5.3). I f t h e s m a l l o r g a n i c c o n t e n t o f s o i l ( t y p i c a l l y 2 % ) i s r e s p o n s i b l e lEor most s o r p t i o n t o s o i l , i t ' s r e a s o n a b l e t o assume t h a t t h e g r e a t m a j o r i t y o f s o r p t i o n t o a c t i v a t e d sludge w i l l be due t o its l a r g e organic c o n t e n t ( t y p i c a l l y 70%). T h i s method i s s u i t a b l e when a n a l y t i c a l . t e s t methods are available for the chemical. It is not appropriate for readily biodegradable or hydrolyzed m a t e r i a l s . However, t h e t e s t c a n be done on stable degradation products. BACK TO MAIN -2- The test measures biosorption at a mixed liquor suspended solids* (MLSS) concentration of 2,000 mg/l, and, if possible, at a test concentration of 1/2 mq/l or 1/2 saturation, whichever is smaller. If test concentrations larger or smaller than 1/2 mg/l must be used, MLSS concentrations should be adjusted up or down proportionately. 1. Materials 1.1 Apparatus 1.1.1 1.1.2 1.1.3 60-70 mesh steel sieve 6 - 50-0-ml Erlenmeyer flasks Boiling water bath 1.1.4 High-speed centrifuge 1.1.5 Spec 20 1.1.6 1.1.7 12-250-m1 plastic centrifuge tubes 7 - 250-ml glass jars with Teflon or aluminum-lined caps. Note: Do not use Teflon-lined caps for evaluations of fluorochemicals unless otherwise stated by requestor. 1.1.8 Pipettes 1.1.9 2-liter volumetric flask 1.1.10 Rotary shaker , 1.1.11 25OC incubator 1.1.12 ASTM thermometer 1.1.13 Graph of MLSS VS 8T at 600 nm (attached) 1.2 Chemicals and Solutions 1.2.1 Deionized (DI) or distilled water. 1.2.2 4 liters activated sludge from an aeration basin at the Metro Wastewater: Treatment System. Use within 48 hrs. of collection. Keep refrigerated ( < 5 O C ) until USE!. *MLSS is the concentration of suspended slids, which are mostly microorganisms, in activated sludge mixed liquor. BACK TO MAIN -3- 1 . 2 . 3 Reagent g r a d e KOH 1.2.4 R e a g e n t g r a d e KH2P04 1.2.5 P r e p a r e a 0.05% p o t a s s i u m p h o s p h a t e b u f f e r by a d d i n g 1 g o f KH2P04 t o 2 l i t e r s of water a n d a d j u s t i n g t o pI 7 w i t h KOH s c l l u t i o n . 1.2.6 2. Procedure Prepare a 1 mg/l s o l u t i o n ( o r a s a t u r a t e d s o l u t i o n , whichever c o n c e n t r a t i o n i s less) of t h e test chemical i n t h e 0.05% phosphate b u f f e r ( S t e p 1 . 2 . 5 ) . Know t h e c o n c e n t r a t i o n of t h i s solution to 3 significant figures. - 2.1 Pass t h e a c t i v a t e d s l u d g e through a 60-70 mesh stainless steel sieve. 2.2 Wash a n d a d j u s t s l u d g e c o n c e n t r a t i o n . 2.2.1 P l a c e s l u d g e i n 250-ml p l a s t i c c e n t r i f u g e t u b e s . (Remember t o b a l a n c e w e i g h t o f opposite centrifuge tubesl) Centrifuge a t 14,000 x G ( 1 0 , 0 0 0 rpm i n I E C B-20A c e n t r i f u g e w i t h 8 7 2 head) for 10 minutes. 2.2.2 Resuspend sludge i n 0.05% potassium phosphate buffer (1.2.5) and centrifuge again as above. 2.2.3 Again, resuspend t h e sludge by shaking with fresh phosphate buffer (1.2.5) and adjust the MLSS concentration t o approximately 4,000 mg/l by making d i l u t i o n s and using t h e attached g r a p h ( A p p e n d i x 1) which compares MLSS a b s o r b a n c e a t 600 nm (of a 2.5 e m l i g h t p a t h . 2.2.4 Determine t h e total suspended matter and total v o l a t i l e s o l i d s i n 2 samples of t h e washed homogeneously resuspended sludge i n accordance with Standard Methods (5.4). 2.3 Place 100.0 m l o f homogeneously suspended washed sludge ( 2 . 2 ) i n t o e a c h of f o u r ( 4 ) 500-ml Erlenmeyer f l a s k s . 2.4 Prepare k i l l e d c e l l s . 2.4.1 P l a c e 2 of t h e f l a s k s from S t e p 2.3 i n a b o i l i n g water b a t h for 15 min. t o k i l l cells. BACK TO MAIN -4- 2.4.2 Bring killed cells back to 25OC in cold water bath or refrigerator before u.sing. 2.5 Add 100.0 ml of the test chemical solution (1.2.6) to one (1) of the 2 flasks containing the killed cells (2.4) and to one (1) of the 2 remaining flasks containing the viable cells (2.3). 2.6 Add 100.0 ml of the phosphate buffer (1.2.5) to the remaining flask of viable cells and to the remaining flask of killed cells. 2.7 Add 100.0 ml of the test chemical solution 1.2.6) and 100 ml of the phosphate buffer (1.2.5) to a 5th 500-ml Erlenmeyer flask. 2.8 Add 200 ml of the phosphate buffer (1.2.5) to a 6th and final 500-ml flask. The 6 flasks have the following composition: (1) 0.5 mg/l (measured to 3 significant figures) test compound + approximately 2,000 mg/l killed activated sludge. (2) approximately 2,000 mg/l killed activated sludge in buffer. (3) 0.5 mg/l test compound + approximately 2,000 mg/l viable activated sludge. (4) Approximately 2,000 mg/l viable activated s l u d g e in buffer. (5) 0.5 mg/l test compound in buffer. (6) Buffer. 2.9 Shake the 6 flasks, prepared in Steps 2.3-2.8, for 1 hour at 25OC. 2.10 Transfer the contents of the Elasks to six (6) 250-ml plastic centrifuge tubes. Centrifuge for 10 min. at 14,000 x g (10,000 rpm in the IEC B-20A centrifuge with 872 head). 2.11 Immediately after the centrifuge stops decant or pipette off the clean supernatant without disturbing the cell pellet. Transfer to glass jars with aluminum foil or Teflon-lined caps. To a 7th jar add 100.0 ml of test chemical solution (1.2.6) and 100.0 ml of buffer (1.2.5) that has not bleen exposed to the plastic centrifuge tube. BACK TO MAIN -5- If immediate analysis is not possible, store sample at 4OC to minimize biodegradation. 2.12 Analyze the water phase of all test and control samples by appropriate methods to determine the concentration of the test compound. 3 . Calculations Calculate the biosorption partition coefficient (K ) between the volatile portions of the sludge and buffer as follows: - ug test chemical/g VSS KP - ug su5strate7ml-Supernatant 3.1 Total chemical in water. TW = Total mass of chemical left in water at sorption equilibrium, ug. WT = Total water used in adsorption study, ml (usually 200 ml). CW = Concentration of chemical in water at sorption equilibrium minus the concentration measured in the chemical-free control water (from flask 6), u9/ml. . 3.2 Total chemical in sludge at sorption equilbrium (assuming no breakdown). TS = Total quantity of chemical sorbed to solids, ug. CC = Final concentration of chemical in sludge free control (from flask 5 ) , ug/ml. WT = Total water used in adsorption s.tudy,ml (usually 200 ml). Tw = Total quantity of chemical left in water, ug (3.1). BACK TO MAIN -6- 3.3 Total Volatile Solids used in Adsorption Study. ST = Total volatile solids used in adsorption study, g. VSS = Volatile suspended solids concentration of sludge added to biosorption study, mg/l (2.2.4). VS = Volume of sludge added to biosorption study, liter (usually 0.-1 liter). 3.4 Biosorption Coefficient (Kp) TS/sT KP = --- CW KP = Biosorption partition coefficient TS = Total quantity of chemical sorbed to solids, ug (3.2). ST = Total volatile solids used in adsorption study, g (3.3). CW = Concentration of chemical in water at sorption equilibrium minus the concentration measured in the chemical-free control, ug/ml. 4 . Presentation of Data 4.1 Test substances 4.2 Initial concentration (concentration in Jar 17 Step 2.11) 4.3 Final concentrations of supernatants from flasks 1 through 6. 4.4 Temperature, OC during shaking (2.9) 4.5 Report K (biosorption coefficient (3.4)) for both the viable syudge and heat-killed sludge. BACK TO MAIN -7- 4.6 Source of sludge, MLSS used in test (usually approximately 2,000 mg/l), VSS used in test (usually 1/2 of that measured). 4.7 PH of all samples 4.8 Time of incubation of test chemical with viable or heat-killed c e l l s (usually 1 hr.). 5. References 5.1 Smith et al, Environmental Pathways of Selected Chemicals in Freshwater Systems, Part I, SRI International, EPA Contract t68-3-2227, EPA-600/7-77113, October 197-7, NTIS PPB-274 548. 5.2 Smith et al, Environmental Pathways of Selected Chemicals in Freshwater Systems, Part 11, EPA-600/7-78074, May 1978. 5.3 ASTM Draft 7, Standard Practicle for Determination of Sorption Constants in Soil and Sediments. 5.4 Standard Methods for the Examination of Water and Wastewater, 14th Edition, American Public Health Association, 1976. EAR/cel BACK TO MAIN 8-19-75 JAP .8 97 .6 I .5 .4 03 .2 MLSS COWNTRATI ON - mg/ 1