Determination of methylene chloride acceptability and its "purification" for ethylenethiourea methodology.

A liquid chromatographic-electrochemical method for the determination of ethylenethiourea (ETU) residues uses methylene chloride in the cleanup. Distilled-in-glass grade methylene chloride produced erratic ETU recoveries ranging from 0 to 106% for vacuum rotary evaporation of the solvent. ETU recoveries were found to be dependent on the bottle of methylene chloride used, not on supplier or lot. When GC2 grade methylene chloride from Burdick & Jackson Laboratories was used, ETU recoveries ranged from 92 to 110%. "Acceptable" ETU recoveries were defined as those values between 90 and 110%. Passing "unacceptable" methylene chloride through a column containing anhydrous sodium sulfate, sodium carbonate, and alumina was found to adequately purify methylene chloride. Treated methylene chloride provided acceptable ETU recoveries for up to 1 month after "purification."

Liquid chromatographic-electrochemical determination of ethylenethiourea in foods by revised official method.

AOAC official method 29.119-29.125 was revised to determine ethylenethiourea (ETU) directly by a liquid chromatographic-electrochemical (LC-EC) determinative technique and to improve ETU recovery. ETU is extracted from food products with a methanol-aqueous sodium acetate solution. A portion of the concentrated filtrate is added to a column of diatomaceous earth, and ETU is eluted with 2% methanol in methylene chloride to separate it from food coextractives, which are retained on the column. The eluate is collected in a siliconized flask and evaporated, the residue is dissolved in water, and 20 microL of solution is injected onto an LC graphitized carbon column. ETU is eluted from the LC column with a mobile phase of acetonitrile-aqueous 0.1M phosphoric acid-water (5 + 25 + 70), and the eluted ETU is detected by using an amperometric electrochemical detector equipped with a gold/mercury working electrode. Recovery data were obtained by fortifying 13 food products with ETU: baked potatoes; canned applesauce, mushrooms, creamed spinach, green beans, spinach, and tomatoes; cooked fresh cabbage and frozen collards; fresh celery and lettuce; grape jelly; and powdered sugar cake donuts. Raw celery was found to cause low ETU recoveries. Average percent recoveries of ETU from the other 12 food products were 92 with a standard deviation of 12 for the low (0.05 and 0.1 ppm) fortification levels and 90 with a standard deviation of 6 for the higher (0.5 and 1 ppm) fortification levels. The limits of quantitation were 0.01 and 0.02 ppm for food products with low and high sugar content, respectively.

Oxidative detection of coulometrically reduced organonitro pesticides in reversed-phase high-performance liquid chromatography.

A high-performance liquid chromatographic system is presented for determination of organonitro pesticides, metabolites and industrial chemicals having a variety of chemical structures. The compounds are chromatographed on a C8 column with an acetonitrile-aqueous ammonium monochloroacetate gradient. The chromatographic responses of the compounds are monitored indirectly by oxidative detection of the coulometrically reduced organonitro functionality by means of a porousgraphite detector. Three nitrophenols (2,4-dinitrophenol, 4,6-dinitro-o-cresol and dinoseb), a dinitrophenyl aliphatic ester (binapacryl) and a nitroaniline (dicloran) were detected at low nanogram levels (3-10 ng for 50% recorder full-scale deflection). This technique overcomes the problems of high background currents and oxygen interference, observed in the direct reductive detection of organonitro compounds.

High-performance liquid chromatographic determination of aryl N-methylcarbamate residues using post-column hydrolysis electrochemical detection.

A high-performance liquid chromatographic technique is reported which selectively detects the phenolic moiety of aryl N-methylcarbamates at low nanogram levels. The carbamates are separated on a C-8 column using an acetonitrile-water gradient mobile phase. The eluted carbamates are hydrolyzed in-line by post-column addition of base, which also serves as electrolyte in the electrochemical (coulometric) detection of the resulting phenols. The technique was tested with six carbamates (bufencarb, carbaryl, carbofuran, 3-hydroxycarbofuran, isoprocarb and methiocarb) and four crops (apples, cabbage, grapes and tomatoes). Optimum detector responses and stability for the carbamates were attained at 0.60 V. Detector response to the carbamates in the presence of crop coextractives was 99% of theoretical with a standard deviation of 2.8% at the 0.05- to 0.1-ppm fortification level. The lower limit of quantitation is 0.01 ppm. Other electrochemical detectors were evaluated for suitability for this work.

Liquid chromatographic determination of N-methylcarbamate insecticides and metabolites in crops. I. Collaborative study.

A liquid chromatographic (LC) multiresidue method for determining residues of N-methylcarbamate insecticides in crops was collaboratively studied in 6 laboratories. Methanol and a mechanical ultrasonic homogenizer are used to extract the carbamates. Water-soluble plant coextractives and nonpolar plant lipid materials are removed from the carbamate residues by liquid-liquid partitioning. Additional crop coextractives (e.g., carotenes, chlorophylls) are removed with a Nuchar SN-silanized Celite column. The carbamate residues are then separated on a reverse phase LC column, using an acetonitrile-water gradient mobile phase. Eluted residues are detected by an in-line post-column fluorometric detection technique. Seven carbamates and 2 carbamate metabolites were included in the collaborative study. Each collaborator determined all the carbamates at 2 levels (approximately 0.05 ppm and United States tolerance) in blind duplicate samples of grapes and potatoes. Fortified and control samples were analyzed. Repeatability coefficients of variation for all the carbamates on the 2 crops averaged 4.7% and ranged from 2.4 to 7.1%. Reproducibility coefficients of variation for all the carbamates on the 2 crops averaged 8.7% and ranged from 5.3 to 12.4%. Accuracy, measured by comparison with fortification values, averaged 95% and ranged from 79 to 103%. The estimated limit of quantitation is 0.01 ppm. The method has been adopted official first action.

Liquid chromatographic determination of N-methylcarbamate insecticides and metabolites in crops. II. Analytical characteristics and residue findings.

Four laboratories obtained 177 carbamate recovery values using a liquid chromatographic method. The average recovery of 11 carbamates (aldicarb, aldicarb sulfone, bufencarb, carbaryl, carbofuran, 3-hydroxy carbofuran, 3-keto carbofuran, methiocarb, methiocarb sulfoxide, methomyl, and oxamyl) from 14 crops was 99% with a coefficient of variation of 8% (0.03-1.8 ppm fortification levels). No statistical difference in recovery was found between oxime and phenyl carbamates, or between parent and metabolite carbamates. Average recovery of aldicarb sulfoxide was 59% due to loss in the liquid-liquid partitioning because of the polarity of this compound. A fifth laboratory contributed 34 carbamate recoveries (average 99%) on table-ready food products for 4 carbamates. Bendiocarb, dioxacarb, isoprocarb, and propoxur are also quantitatively recovered through the method. Previously reported carbamate and noncarbamate recovery data are also discussed. In the Food and Drug Administration's (FDA) analysis of 319 samples (mainly crops), 86 (27%) were found to contain residues of carbamate insecticides and/or toxic carbamate metabolites. Carbaryl and methomyl were the most common carbamate residues found on the food products excluding the aldicarb sulfone and sulfoxide residues found on potatoes. In one FDA Total Diet Program "market basket", 11 of 69 table-ready food commodities contained from 0.005 to 0.094 ppm carbamate residues. Carbaryl was the most prevalent residue. Several laboratories reported adverse effects on the determinative system when inadequately purified reagents were used.

Determination of coumaphos and its oxygen analog in eggs and milk by using a multiresidue method with liquid chromatographic quantitation and capillary gas chromatographic/mass spectrometric confirmation.

A multiresidue method for carbamate insecticides was adapted for the determination of coumaphos and its oxygen analog in eggs and milk. Eggs were extracted with acetonitrile and milk was extracted with acetone. Co-extractives were removed using liquid partitioning and charcoal column procedures described in the carbamate method. Coumaphos and its oxygen analog were determined by using a high performance liquid chromatograph equipped with a fluorescence detector. Recovery studies were performed for the 2 compounds at levels of 0.01 and 0.10 ppm in eggs and 0.01 and 0.02 ppm in milk. Overall average recovery was 100% (range 95-109%). In a trial of the method by another laboratory, the recovery of coumaphos and its oxygen analog from milk averaged 87 and 96%, respectively. Data are presented on the capillary gas chromatographic/mass spectrometric confirmation of coumaphos residues.

Applicability of a multiresidue method and high performance liquid chromatography for determining quinomethionate in apples and oranges.

The AOAC multiresidue method for nonpolar pesticide residues in nonfatty foods has been coupled with a high performance liquid chromatographic (HPLC)-fluorometric system for determining quinomethionate residues on apples and oranges. Quinomethionate is extracted with acetonitrile, and coextractives are removed with liquid-liquid partitioning and Florisil adsorbent using the AOAC multipesticide residue method for nonfatty foods. The quinomethionate fraction is then chromatographed on an HPLC octyl-bonded column and detected in-line with a fluorescence detector using 362 nm excitation and 395 nm emission. Recovery studies were conducted with apples fortified with quinomethionate at 0.05 ppm and oranges at 0.05 and 0.5 ppm. The recoveries averaged 100% (range 92-108) at the 0.05 ppm fortification level and 102% (range 93-110) at the 0.5 ppm level.

Applicability of a carbamate insecticide multiresidue method for determining additional types of pesticides in fruits and vegetables.

Several fruits and vegetables were fortified at a low (0.02-0.5 ppm) and at a high (0.1-5 ppm) level with pesticides and with a synergist, and recoveries were determined. Analyses were performed by using 3 steps of a multiresidue method for determining N-methylcarbamates in crops: methanol extraction followed by removal of plant co-extractives by solvent partitioning and chromatography with a charcoal-silanized Celite column. Eleven compounds were determined by using a high performance liquid chromatograph equipped with a reverse phase column and a fluorescence detector. Twelve additional compounds were determined by using a gas-liquid chromatograph equipped with a nonpolar packed column and an electron capture or flame photometric detector. Recoveries of 10 pesticides (azinphos ethyl, azinphos methyl, azinphos methyl oxygen analog, carbaryl, carbofuran, naphthalene acetamide, naphthalene acetic acid methyl ester, napropamide, phosalone, and phosalone oxygen analog) and the synergist piperonyl butoxide, which were determined by high performance liquid chromatography, averaged 100% (range 86-117) at the low fortification level and 102% (range 93-115) at the high fortification level. Quantitative recovery of naphthalene acetamide through the method required that an additional portion of eluting solution be passed through the charcoal column. Recoveries of 7 additional pesticides (dimethoate, malathion, methyl parathion, mevinphos, parathion, phorate oxygen analog, and pronamide), which were determined by gas-liquid chromatography (GLC), averaged 108% (range 100-120) at the low fortification level and 107% (range 99-122) at the high fortification level. DDT, diazinon, dieldrin, phorate, and pirimiphos ethyl, which were determined by GLC, were not quantitatively recovered.

Influence of various solvent-water mixtures on the extraction of dieldrin and methomyl residues from radishes.

The effect of organic solvent/water ratios on the extraction of field-incurred residues of dieldrin and methomyl from radishes was determined. 14C-Dieldrin and 14C-methomyl were applied separately to radishes in commercial formulations at rates of 0.2 and 0.9 kg/ha, respectively. Fourteen days post-application, the radishes were harvested and fresh root tissues were extracted using a Polytron homogenizer. Acetone, acetonitrile, and methanol containing 0, 10, 20, 30, 40, and 50% water were used as extraction solvents. For methomyl residues, the optimum water content of acetonitrile-water extraction mixtures was 40-50%; less than 40% water reduced the ability of acetonitrile to extract carbon-14. Methanol and acetone were nearly as efficient as 50% acetonitrile-water and were apparently not influenced by solvent/water ratios. For dieldrin, low water content slightly reduced the extraction efficiency of acetonitrile, with the optimum water content also being 40-50%. Percentage of water appeared to have little overall effect on methanol extraction efficiency. The extraction efficiency of acetone was lower than that of the other 2 solvents, and this effect was independent of the acetone/water ratio. Approximately 20% of the 14C-dieldrin residue was bound to radish roots 14 days post-application.

Degradation of methomyl residues in frozen strawberries.

14C-Methomyl (S-methyl N-[(methylcarbamoyl)-oxy]thioacetimidate) suspended in a commercial formulation was sprayed on strawberries. At 7 and 14 days post-application, mature berries were harvested in chopped. Fresh chopped berries were extracted with methanol using a Polytron homogenizer, and the crop marc was subsequently leached with methanol, freeze-dried, and re-extracted. Portions of the fresh chopped fruit were also frozen at-20 degrees C for 1, 30, and 102 days before extraction with methanol using a Polytron homogenizer, followed by subsequent leaching of the crop marc. For fresh berries, 93.7 and 88.9% of the 14C present at harvest was extractable at 7 and 14 days post-application, respectively. Based on thin layer chromatographic analysis, methomyl represented 83.5 and 68.2% of the 14C extracted from fresh berries at 7 and 14 days, respectively. Freezing of strawberries reduced extraction efficiency and appeared to degrade methomyl. At 7 and 14 days post-application, averages of 80.9 and 77.1%, respectively, of the 14C present were extractable after frozen storage regardless of storage time. Furthermore, only about 10% of the extracted 14C was methomyl; the remainder was of unknown composition.

Extraction efficiencies for pesticides in crops: 14C-benomyl extraction from mustard green and radishes.

14C-labeled benomyl [methyl 1-(butylcarbamoyl)-2-benzimidazolecarbamate] suspended in a commercial benomyl formulation was sprayed on mustard greens and radishes. At 3 intervals after application, the crops were extracted with methanol, acetonitrile, or acetone. Crops were either blended and leached or repetitively blended followed by Soxhlet extraction. Essentially all of the extractable radioactivity was removed by blending. The 14C was more difficult to extract from radishes than from mustard greens as time increased. Respective percentages of 14C extracted at 1, 7, and 14 days were 99, 98, and 97 from mustard greens and 96, 88, and 79 from radishes. Methanol exhibited the highest extraction efficiency, and the blend-Soxhlet process was better than the blend-leach process. Thin layer chromatography of the organic-soluble extracts indicated that the majority of 14C was recovered as methyl 2-benzimidazole carbamate (MBC), a breakdown product of benomyl. Acid hydrolysis of the extracted tissues released 30--50% of the residual 14C.