SECTION 302 Pesticide Analytical Manual Vol I ALTERNAT/VE E3 EXTRACTION WTTH ACETONE, REMOVAL OF WATER WITH 25 G HYDROMATRX Reference PAM 1 232.4(Luke procedure)Acetone Filtrate, "LIB 3613, FDA, Rockville, M03 Palmer, R.E., and Hopper, M L.(Nov. 1991)"Miniaturized Solid Phase Partitio Column for Determination of Organochlorine and Organophosphate Pesticides wit Principles Smaller size column of Hydromatrix reduces solvent use by 40% over E2, while still removing water from same amount of extract. However, solution eluting from 25 Hydromatrix column may be cloudy, probably from a small amount of water; this disappears during concentration. The 25 g column may also have a shorter lifetime than the 40 g column Results using the 25 g column may be somewhat less reliable for certain chemicals; e.g., p,p'-dicofol and dicloran are recovered less reproducibly, and >0. 4 ppm methamidophos may be only partially recovered; elution with 300 mL methylene chloride permits complete recovery of the latter. Directions Follow directions of E2, except Prepare Hydromatrix column from 25 g material instead of 40 g Prewash Hydromatrix column with 100 mL acetone followed by 100 mL methylene chloride immediately before each use After transferring 40 mL filtered acetone extract to top of column, elute with 25, 25, and 150 mL methylene chloride, instead of volumes used in Because amount of original sample and amount of filtered acetone extract transferred to Hydromatrix column are the same as in E2, mg sample equivalent is the same as E2. 30a-12 230
SECTION 302 Pesticide Analytical Manual Vol. I 302–12 Transmittal No. 2000-1 (10/1999) Form FDA 2905a (6/92) ALTERNATIVE: E3 EXTRACTION WITH ACETONE, REMOVAL OF WATER WITH 25 G HYDROMATRIX Reference Palmer, R.E., and Hopper, M.L. (Nov. 1991) “Miniaturized Solid Phase Partition Column for Determination of Organochlorine and Organophosphate Pesticides with PAM I 232.4 (Luke procedure) Acetone Filtrate,” LIB 3613, FDA, Rockville, MD Principles Smaller size column of Hydromatrix reduces solvent use by 40% over E2, while still removing water from same amount of extract. However, solution eluting from 25 g Hydromatrix column may be cloudy, probably from a small amount of water; this disappears during concentration. The 25 g column may also have a shorter lifetime than the 40 g column. Results using the 25 g column may be somewhat less reliable for certain chemicals; e.g., p,p'-dicofol and dicloran are recovered less reproducibly, and >0.4 ppm methamidophos may be only partially recovered; elution with 300 mL methylene chloride permits complete recovery of the latter. Directions • Follow directions of E2, except: – Prepare Hydromatrix column from 25 g material instead of 40 g. – Prewash Hydromatrix column with 100 mL acetone followed by 100 mL methylene chloride immediately before each use. – After transferring 40 mL filtered acetone extract to top of column, elute with 25, 25, and 150 mL methylene chloride, instead of volumes used in E2. – Because amount of original sample and amount of filtered acetone extract transferred to Hydromatrix column are the same as in E2, mg sample equivalent is the same as E2
Pesticide Analytical Manual Vol. I SECTION 302 E4 EXTRACTION WITH WATER/ACETONE, LIQUID-LIQUID PARTITIONING WITH PETROLEUM ETHER/METHYLENE CHLORIDE Reference Luke, M.A., and Doose, GM.(1983)Bull. Environ Contam. Toxicol. 30, 110-116 Principles Low moisture nonfatty sample is blended with 35% water/acetone and filtered; the presence of water in the extractant facilitates extraction of residues from the dry productand dilutes co-extractives Most nonionic residues are extracted into aqueous acetone solution Residues are transferred from aqueous acetone to organic solvent methylene chloride/petroleum ether by partitioning, with salt added to the aqueous layer after the first partitioning to aid transfer Appar blender, high speed; explosion-proof Waring Blendor, I qt jar Buchner funnel (Buchner), porcelain, 12 cm diameter filter paper, Shark Skin, to fit Buchner long-stemmed funnel, glass, 4"diameter grinder, suitable for reducing dry products to <20 mesh Kuderna-Danish concentrator(K-D),500 mL, with Snyder column, two-ball micro-Snyder column, graduated receiving flask separatory funnel(separator), I L R acetone, distilled from all-glass apparatus boiling chips, 20-30 mesh carborundum(optional) glass wool, Pyrex; see Section 204 for handling directions methylene chloride, distilled from all-glass apparatus petroleum ether, distilled from all-glass apparatus sodium chloride, reagent grade sodium sulfate, anhydrous granular, reagent grade: see Section 204 for handling directions 35%(v/v) water/acetone Prewash filter paper with acetone to remove artifact Grind sample containing <10% fat or oil to <20 mesh Weigh 15 g ground sample into blender jar, add 950 mL 35% water/ acetone, and blend 2 min at high speed Filter with suction through 12 cm Buchner fitted with Shark Skin paper collect extract in 500 mL suction flask. Filtration is normally complete in <l min Continuation of vacuum for excessive period can reduce volume of extract and cause error in calculation Place 80 mL sample extract in I L separator containing 100 mL methyler chloride. Add 100 mL petroleum ether and shake vigorously I min. ne FDA 2905a(6/92) 302-13
Transmittal No. 2000-1 (10/1999) Form FDA 2905a (6/92) 302–13 Pesticide Analytical Manual Vol. I SECTION 302 E4 EXTRACTION WITH WATER/ACETONE, LIQUID-LIQUID PARTITIONING WITH PETROLEUM ETHER/METHYLENE CHLORIDE Reference Luke, M.A., and Doose, G.M. (1983) Bull. Environ. Contam. Toxicol. 30, 110-116 Principles Low moisture nonfatty sample is blended with 35% water/acetone and filtered; the presence of water in the extractant facilitates extraction of residues from the dry product and dilutes co-extractives. Most nonionic residues are extracted into aqueous acetone solution. Residues are transferred from aqueous acetone to organic solvent methylene chloride/petroleum ether by partitioning, with salt added to the aqueous layer after the first partitioning to aid transfer. Apparatus blender, high speed; explosion-proof Waring Blendor, 1 qt jar Büchner funnel (Büchner), porcelain, 12 cm diameter filter paper, Shark Skin®, to fit Büchner long-stemmed funnel, glass, 4" diameter grinder, suitable for reducing dry products to <20 mesh Kuderna-Danish concentrator (K-D), 500 mL, with Snyder column, two-ball micro-Snyder column, graduated receiving flask separatory funnel (separator), 1 L Reagents acetone, distilled from all-glass apparatus boiling chips, 20-30 mesh carborundum (optional) glass wool, Pyrex; see Section 204 for handling directions methylene chloride, distilled from all-glass apparatus petroleum ether, distilled from all-glass apparatus sodium chloride, reagent grade sodium sulfate, anhydrous, granular, reagent grade; see Section 204 for handling directions 35% (v/v) water/acetone Directions • Prewash filter paper with acetone to remove artifacts. • Grind sample containing <10% fat or oil to <20 mesh. • Weigh 15 g ground sample into blender jar, add 350 mL 35% water/ acetone, and blend 2 min at high speed. • Filter with suction through 12 cm Büchner fitted with Shark Skin® paper; collect extract in 500 mL suction flask. Filtration is normally complete in <1 min. Continuation of vacuum for excessive period can reduce volume of extract and cause error in calculation. • Place 80 mL sample extract in 1 L separator containing 100 mL methylene chloride. Add 100 mL petroleum ether and shake vigorously 1 min
SECTION 302 Pesticide Analytical Manual Vol I Transfer lower aqueous layer to second I L separator Dry upper organic layer of first separator by passing through about 1.5 sodium sulfate supported on washed glass wool in 4"funnel, collecting in K-D.(If extract will be cleaned up directly with C3, charcoal/Celite column, collect in vacuum rotary evaporator flask. To separator with aqueous phase, add 7 g sodium chloride and shake vigorously 30 sec until most of the sodium chloride is dissolved Add 100 mL methylene chloride, shake l min, and dry lower organic phase through same sodium sulfate. Extract aqueous phase with additional 100 mL methylene chloride and dry as above. Rinse sodium sulfate with about 50 mL methylene chloride (If extractwill be cleaned up directly with C3, proceed to concentration step described there instead of evaporating in K-D as follows. Add boiling chips to K-D and concentrate solvent in K-D; start evaporation slowly by placing only receiver tube into steam. After 100-150 mL has evaporated, concentrator may be exposed to more steam. When liquid level in hot concentrator tube is about 2 mL, add 100 mL petroleum ether through Snyder column and reconcentrate to about 2 mL. Add 50 L petroleum ether and repeat concentration step. Add 20 mL acetone, and reconcentrate to about 2 mL. Do not allow solution to go to dryness during any of the concentration steps. Adjust volume of extract to suitable definite Calculate equivalent sample weight in final solution mg sample equivalent =15 L final extract 350 mL final volume where 15=g sample analyzed 80= mL filtered extract taken for liquid-liquid partitioning Thus, when final extract volume is 2 mL, each uL contains 1 1.7 mg sample equivale 15 3502 LL final extract Extract may be suitable, as is, for determination by glC with selective detectors(e.g, DG2, DG3).If co-extractives interfere with determination or adversely affect chromatography, clean up extract with Cl, C2, or C5 prior to determination Clean up extract with Cl or C5 prior to determination by electron capture DGl, DG7, etc. )or flame ionization detectors(DG6) Clean up extract with CB or C4 prior to determination by DLI for N-methylcarbamates 302-14 230
SECTION 302 Pesticide Analytical Manual Vol. I 302–14 Transmittal No. 2000-1 (10/1999) Form FDA 2905a (6/92) • Transfer lower aqueous layer to second 1 L separator. • Dry upper organic layer of first separator by passing through about 1.5" sodium sulfate supported on washed glass wool in 4" funnel, collecting in K-D. (If extract will be cleaned up directly with C3, charcoal/Celite column, collect in vacuum rotary evaporator flask.) • To separator with aqueous phase, add 7 g sodium chloride and shake vigorously 30 sec until most of the sodium chloride is dissolved. • Add 100 mL methylene chloride, shake 1 min, and dry lower organic phase through same sodium sulfate. • Extract aqueous phase with additional 100 mL methylene chloride and dry as above. Rinse sodium sulfate with about 50 mL methylene chloride. (If extract will be cleaned up directly with C3, proceed to concentration step described there instead of evaporating in K-D as follows.) • Add boiling chips to K-D and concentrate solvent in K-D; start evaporation slowly by placing only receiver tube into steam. After 100-150 mL has evaporated, concentrator may be exposed to more steam. When liquid level in hot concentrator tube is about 2 mL, add 100 mL petroleum ether through Snyder column and reconcentrate to about 2 mL. Add 50 mL petroleum ether and repeat concentration step. Add 20 mL acetone, and reconcentrate to about 2 mL. Do not allow solution to go to dryness during any of the concentration steps. Adjust volume of extract to suitable definite volume with acetone. • Calculate equivalent sample weight in final solution: mg sample equivalent µL final extract = 15 × 80 350 1 mL final volume × where: 15 = g sample analyzed 80 = mL filtered extract taken for liquid-liquid partitioning 350 = mL water/acetone blended with 15 g sample Thus, when final extract volume is 2 mL, each µL contains: 1.7 mg sample equivalent µL final extract 15 × 80 350 1 2 × = • Extract may be suitable, as is, for determination by GLC with selective detectors (e.g., DG2, DG3). If co-extractives interfere with determination or adversely affect chromatography, clean up extract with C1, C2, or C5 prior to determination. • Clean up extract with C1 or C5 prior to determination by electron capture (DG1, DG7, etc.) or flame ionization detectors (DG6). Clean up extract with C3 or C4 prior to determination by DL1 for N-methylcarbamates
Pesticide Analytical Manual Vol. I SECTION 302 ALTERNATIVE E5 EXTRACTION WITH ACETONE. LIQUID-L/QUID PARTITIONING WITH ACETONE/METHYLENE CHLORIDE Reference Luke, M. A, and Doose, G. M.(1983)Bull. Environ Contam. Toxicol. 30, 110-116 Principle ether/dichloromethane is used in partitioning Better recoveries are obtained when acetone is substituted for petroleum ether. Transfer of polar pesticides from the aqueous phase to the organic layer is further facilitated by adding sodium chloride before, rather than after, the first partitioning step Directions Follow directions of El through blending and filtering. Then Place 80 mL sample extract in I L separator, and add 100 mL acetone, 100 mLmethylene chloride, and 7 sodium chloride. Shake vigorously I min Transfer lower aqueous layer to second I L separator Dry upper organic layer of first separator by passing through about 1.5 sodium sulfate supported on washed glass wool in 4 "funnel, collectin in K-D.(If extract will be cleaned up directly with C3, charcoal/ silanized Celite column, collect in vacuum rotary evaporator flask. Add 100 mL methylene chloride, shake 1 min, and dry lower organic hase through same sodium sulfate Continue as in El, "Extract aqueous phase with additional 100 mL methylene chloride ALTERNAT/VE E6 EXTRACTION WITH WATER/ACETONE, LIQUID-LIQUID PARTITIONING WITH ACETONE/METHYLENE CHLORIDE Reference Luke, M.A., and Doose, G. M.(198%)Bull. Environ Contam. Toxicol. 30, 110-116 Principle Polar pesticides such as methamidophos exhibit variable recoveries when petroleum ether/methylene chloride is used in partitioning. Better recoveries are obtained when acetone is substituted for petroleum ether. Transfer of polar pesticides from the aqueous phase to the organic layer is further facilitated by adding sodium chloride before, rather than after, the first partitioning step 15
Transmittal No. 2000-1 (10/1999) Form FDA 2905a (6/92) 302–15 Pesticide Analytical Manual Vol. I SECTION 302 ALTERNATIVE: E5 EXTRACTION WITH ACETONE, LIQUID-LIQUID PARTITIONING WITH ACETONE/METHYLENE CHLORIDE Reference Luke, M. A., and Doose, G. M. (1983) Bull. Environ. Contam. Toxicol. 30, 110-116 Principle Polar pesticides such as methamidophos exhibit variable recoveries when petroleum ether/dichloromethane is used in partitioning. Better recoveries are obtained when acetone is substituted for petroleum ether. Transfer of polar pesticides from the aqueous phase to the organic layer is further facilitated by adding sodium chloride before, rather than after, the first partitioning step. Directions • Follow directions of E1 through blending and filtering. Then: – Place 80 mL sample extract in 1 L separator, and add 100 mL acetone, 100 mL methylene chloride, and 7 g sodium chloride. Shake vigorously 1 min. – Transfer lower aqueous layer to second 1 L separator. – Dry upper organic layer of first separator by passing through about 1.5" sodium sulfate supported on washed glass wool in 4" funnel, collecting in K-D. (If extract will be cleaned up directly with C3, charcoal/ silanized Celite column, collect in vacuum rotary evaporator flask.) – Add 100 mL methylene chloride, shake 1 min, and dry lower organic phase through same sodium sulfate. • Continue as in E1, “Extract aqueous phase with additional 100 mL methylene chloride...” ALTERNATIVE: E6 EXTRACTION WITH WATER/ACETONE, LIQUID-LIQUID PARTITIONING WITH ACETONE/METHYLENE CHLORIDE Reference Luke, M. A., and Doose, G. M. (1983) Bull. Environ. Contam. Toxicol. 30, 110-116 Principle Polar pesticides such as methamidophos exhibit variable recoveries when petroleum ether/methylene chloride is used in partitioning. Better recoveries are obtained when acetone is substituted for petroleum ether. Transfer of polar pesticides from the aqueous phase to the organic layer is further facilitated by adding sodium chloride before, rather than after, the first partitioning step
SECTION 302 Pesticide Analytical Manual Vol I Directions Follow directions of e4 through blending and filtering. Then: Place 80 mL sample extract in I L separator containing 100 mL methyl ene chloride. Add 100 mL acetone and 7 g sodium chloride and shake vigorously l min. Transfer lower aqueous layer to second I L separator Dry upper organic layer of first separator by passing through about 1.5 sodium sulfate supported on washed glass wool in 4 "funnel, collecting in K-D. (If extract will be cleaned up directly with C%, charcoal/ silanized Celite column, collect in vacuum rotary evaporator flask. Add 100 mL methylene chloride, shake I min, and dry lower organic phase through same sodium sulfate Continue as in E4, "Extract aqueous phase with additional 100 mL methyl ene chloride 302-16 230
SECTION 302 Pesticide Analytical Manual Vol. I 302–16 Transmittal No. 2000-1 (10/1999) Form FDA 2905a (6/92) Directions • Follow directions of E4 through blending and filtering. Then: – Place 80 mL sample extract in 1 L separator containing 100 mL methylene chloride. Add 100 mL acetone and 7 g sodium chloride and shake vigorously 1 min. – Transfer lower aqueous layer to second 1 L separator. – Dry upper organic layer of first separator by passing through about 1.5" sodium sulfate supported on washed glass wool in 4" funnel, collecting in K-D. (If extract will be cleaned up directly with C3, charcoal/silanized Celite column, collect in vacuum rotary evaporator flask.) – Add 100 mL methylene chloride, shake 1 min, and dry lower organic phase through same sodium sulfate. • Continue as in E4, “Extract aqueous phase with additional 100 mL methyl ene chloride