Electron capture gas-liquid chromatographic method for the simultaneous analysis of 2,4-D, dicamba, and mecoprop residues in soil, wheat, and barley.

A method has been developed for the simultaneous analysis of 2,4-D (2,4-dichlorophenoxy-acetic acid), dicamba (2-methoxy-3,6-dichloro-benzoic acid), and mecoprop (MCPP; 2-[(4-chloro-o-tolyl) oxy] propionic acid) residues in soil, wheat, and barley. Soil and crop samples are extracted with acidic acetone and methanol, respectively. The extracts in diethyl ether are esterified with diazomethane and cleaned up by passing through a Florisil column. Extracts are analyzed by gas-liquid chromatography, using an electron capture detector to determine 2,4-D and dicamba residues. Mecoprop in the extract is not detected at low levels of concentration. However, bromination of the extract increases the response of the electron capture detector to mecoprop. The method is sensitive to about 0.05 ppm 2,4-D and dicamba and 0.5 ppm mecoprop. Recoveries of these 3 herbicides added to soil, wheat, and barley samples at 0.05, 0.1, 0.5, and 1.0 ppm levels were between 65 and 93%. The method was used for the simultaneous analysis of 2,4-D, dicamba, and mecoprop residues in wheat, barley, and soil samples obtained from fields sprayed with the herbicide formulation Kil-Mor.     

Liquid chromatographic determination of lasalocid sodium in chicken skin: interlaboratory study

The U.S. Food and Drug Administration (FDA) sponsored an interlaboratory study of a liquid chromatographic determinative procedure for lasalocid sodium in chicken skin with adhering fat. Four laboratories analyzed 35 dosed tissue samples and 82 fortified tissue samples containing lasalocid at levels ranging from 0.1 to 0.6 ppm. Samples were homogenized with acetonitrile, washed with hexane, and partitioned into the mobile phase prior to analysis liquid chromatography with fluorescence detection. The results of the interlaboratory study showed good reproducibility for fortified samples. Fortification levels, average recoveries, and interlaboratory percent coefficients of variation were as follows: 0.6 ppm, 0.57 ppm, and 9.7; 0.3 ppm, 0.25 ppm, and 9.1; and 0.15 ppm, 0.14 ppm, and 7.0, respectively. Data for analysis of the dosed tissue also showed good agreement among the laboratories.     

Improved cleanup for gas chromatographic determination of propiconazole residues in soil, wheat grain, straw, and leaves

A simple and sensitive method is described for determination of propiconazole, a new type of broad-spectrum systemic fungicide, in soil, wheat grain, straw, and leaves. Pesticide residues in or on grain and green plant materials are extracted with methanol (or a mixture of methanol and water (4 + 1), for soil), partitioned into methylene chloride, and cleaned up on an alumina column for grain and soil or an activated charcoal column for green plant materials. The amount of residue is quantitatively measured by gas chromatography using an alkali flame ionization detector in the nitrogen-sensitive mode. Recoveries from soil, grain, and green plant materials fortified at 0.1-5 mg/kg are better than 80%. The practical detection limits of this method are 0.01 mg/kg in grain and soil and 0.02 mg/kg in green plant materials.     

Gas-liquid chromatographic determination of adipic acid in crackling candy and soft drinks.

A procedure was developed for the simple and rapid determination of adipic acid in crackling candy and also in soft drinks. An alkaline solution of sample was extracted with ethyl ether to remove fatty substances, and H2SO4 was added to water layer to adjust the pH to less than 2. The acidified layer was saturated with NaCl and then extracted with ether. After drying, the ether layer was concentrated and the adipic acid in the concentrate was methylated using the diazomethane methograph equipped with a flame ionization detector. Recovery of adipic acid from crackling candy and from 2 kinds of soft drinks that had been fortified at the 200 ppm level was 96%. An interlaboratory test was carried out on the determination of adipic acid in orange soft drink. The results obtained by 6 laboratories were between 91 and 100% compared with the theoretical value.     

Gas chromatographic determination of captan, folpet, and captafol residues in tomatoes, cucumbers, and apples using a wide-bore capillary column: interlaboratory study.

An interlaboratory study of the determination of captan, folpet, and captafol in tomatoes, cucumbers, and apples was conducted by 4 laboratories using wide-bore capillary column gas chromatography with electron capture detection. The 3 fungicides were determined using the Luke et al. multiresidue method modified to include additional solvent elution in the optional Florisil column cleanup step used with this method. The crops were fortified with each fungicide at 3 levels per crop. Mean recoveries ranged from 86.2% for a 25.1 ppm level of captan in apples to 115.4% for a 0.288 ppm level of captafol in apples. Interlaboratory coefficients of variation ranged from 3.4% (24.7 ppm folpet) to 9.7% (0.243 ppm captafol) for tomatoes; from 2.8% (2.0 ppm captafol) to 8.2% (24.8 ppm captan) for cucumbers; and from 1.5% (0.234 ppm folpet) to 22.1% (0.266 ppm captafol) for apples.     

The Luke et al. method for determining multipesticide residues in fruits and vegetables: collaborative study

Ten laboratories analyzed unfortified and fortified samples of lettuce, tomatoes, and strawberries for organochlorine and organophosphorus pesticides by applicable portions of the comprehensive multipesticide method of Luke et al. The 3 crops were fortified with 6 pesticides, alpha-BHC, dieldrin, chlorpyrifos, acephate, omethoate, and monocrotophos, each at 3 levels per crop. Included in the 54 fortifications were 16 pairs of blind duplicates: same pesticide, crop, and level. Recoveries were calculated by area comparisons with known reference materials, using the responses obtained from 2 separate element-specific gas chromatographic (GC) systems. The organochlorine pesticides were chromatographed on a methyl silicone column and detected with a Hall 700A electrolytic conductivity detector, and the organophosphorus pesticides were determined with a flame photometric detector after being chromatographed on a specified DEGS column material. Chlorpyrifos was quantitated on both GC systems. Mean recoveries ranged from 82.6% for acephate fortified at 0.5000 ppm in strawberries to 118.1% for 0.0636 ppm fortification of chlorpyrifos in lettuce. Interlaboratory coefficients of variation ranged from 4.0% for 0.6360 ppm fortification of chlorpyrifos in tomatoes to 17.8% for the 0.0636 ppm chlorpyrifos level in lettuce. The procedure features essentially no cleanup before GC and proved comparable to existing multiresidue methods for pesticides of the class types studied, as evidenced by the intra- and interlaboratory measurements of precision and recoveries obtained. The method with the 2 GC systems has been adopted official first action.     

Comparative degradation of [14C]-2,4-dichlorophenoxyacetic acid in wheat and potato after Foliar application and in wheat, radish, lettuce, and apple after soil application

The fate of 2,4-dichlorophenoxyacetic acid (2,4-D) applied foliarly as the 2-ethylhexyl ester (EHE) to wheat and potatoes, to the soil as the dimethylamine (DMA) salt under apple tree canopies, and preplant as the free acid for wheat, lettuce, and radish was studied to evaluate metabolic pathways. Crop fractions analyzed for (14)C residues included wheat forage, straw, and grain; potato vine and tubers; and apple fruit. The primary metabolic pathway for foliar application in wheat is ester hydrolysis followed by the formation of base-labile 2,4-D conjugates. A less significant pathway for 2,4-D in wheat was ring hydroxylation to give NIH-shift products 2,5-dichloro-4-hydroxyphenoxyacetic acid (4-OH-2,5-D), 4-OH-2,3-D, and 5-OH-2,4-D both free and as acid-labile conjugates. The primary metabolic pathway in potato was again ester hydrolysis. 2,4-D acid was further transformed to 4-chlorophenoxyacetic acid and 4-OH-2,5-D. For the soil applications, (14)C residues in the crops were low, and characterization of the (14)C residues indicated association with or incorporation into the biochemical matrix of the tissue. The degradative pathways observed in wheat are similar to those characterized in other intact plant studies but differ from those in studies in wheat cell suspension culture in that no amino acid conjugates were observed.     

Determination of N-nitrosodimethylamine levels in some Canadian 2,4-D amine formulations

Levels of N-nitrosodimethylamine (NDMA) were determined in 112 samples of 2,4-dichlorophenoxyacetic acid, (2,4-D), formulated as the dimethylamine salt, collected over a 2 year period from products on the Canadian market. A sample aliquot is partitioned with dichloromethane, and the co-extracted dimethylamine is removed by cleanup on a silica gel column. The eluates containing NDMA are concentrated, an internal standard of N-nitrosodipropylamine is added, and nitrosamine levels are determined using a gas chromatograph interfaced with a thermal energy analyzer. Recoveries of NDMA and N-nitrosodiethylamine spiked into samples were 103 +/- 16 and 96.3 +/- 9.8%, respectively. Of the 112 samples analyzed, 92 were below 1 part per million (ppm) relative to the amount of 2,4-D in the samples, 16 were between 1 and 5 ppm, and 4 were greater than 5 ppm. The gas chromatographic column used is compared to a conventional packing material for volatile nitrosamine analysis. Formation of NDMA during cleanup and analysis was shown not to occur.     

Atomic absorption determination of tin in foods: collaborative study

Samples of green beans, applesauce, and a fruit juice were fortified with tin at 3 levels. Collaborators were asked to digest the samples, using HNO3-H2SO4, add methanol to enhance the absorption signal, and aspirate directly, using a nitrous oxide-acetylene flame. Results were received from 8 laboratories including 4 from Europe. However, only 6 laboratories used the prescribed methodology. All results were considered acceptable. The method has been adopted as interim official first action.     

Gas-liquid chromatographic determination of captan and two metabolites in milk and meat

A gas-liquid chromatographic (GLC) method has been developed for the determination of captan (N-trichloromethylthio-4-cyclohexene-1,2-dicarboximide) and 2 metabolites, tetra-hydrophthalimide (THPI) and tetrahydrophthalamic acid (THPMA), in milk and meat. The sample is extracted with ethyl acetate and is cleaned up by acetonitrile partition and silica gel chromatography where captan, THPI, and THPMA are separated. Captan is directly determined by GLC. THPI and THPMA are separately derivatized in an acetone solution of pentafluorobenzyl bromide. The resultant derivatives are purified separately on an Al2O3 column and quantitated by GLC, using an electron capture detector. Recoveries from milk samples fortified at 0.02-10 ppm ranged from 71 to 102%; recoveries from meat samples fortified at 0.04-10 ppm ranged from 75 to 99%.     

Gas-liquid chromatographic determination of dehydroacetic acid in squash and wine

A gas-liquid chromatographic procedure has been developed for the determination of the preservative dehydroacetic acid (DHA) in frozen cut squash and white wine. The cleanup procedure uses the acidic character of DHA to enhance separation from interfering substances. Recoveries were 93-104% from squash fortified at 30, 65, and 130 ppm and 96-99% from wine fortified at 50, 100, and 200 ppm. The method can be used to establish that the amount of DHA used in squash does not exceed the permissible level of 65 ppm.     

Uniformly (14)C-ring-labeled 2,4-dichlorophenoxyacetic acid: a metabolism study in bluegill sunfish, Lepomis macrochirus.

The fate of 2,4-dichlorophenoxyacetic acid (2,4-D), a mixture of [phenyl(U)-(14)C]-2,4-D and unlabeled 2,4-D, in bluegill sunfish was investigated after exposure to approximately 11 ppm under static conditions for 4 days. Total radioactive residues (TRR) in whole fish increased from 0.41 ppm on day 1 to 0.60 ppm on day 3. TRR levels in fillet (edible) and viscera (nonedible) of treated fish on day 4 were 0.41 and 1.9 ppm, respectively. Most residues in both matrices were acetonitrile soluble; small amounts were hexane soluble or unextractable with solvents. Acid and base hydrolyses with ethyl acetate partitioning were used to release the fillet unextractable residues. The identification of 2,4-D and 2,4-dichlorophenol (2,4-DCP) in the fillet was conclusively confirmed by GC-MS analysis. On the basis of the experimental data from this study, a metabolic pathway for 2,4-D in bluegill sunfish in which the 2,4-D is metabolized to 2,4-DCP and conjugates of 2,4-D and 2,4-DCP is proposed.     

Liquid chromatographic method for multiresidue determination of benzimidazoles in beef liver and muscle: collaborative study

A liquid chromatographic method for determination of thiabendazole, 5-hydroxythiabendazole, oxfendazole, mebendazole (MBZ), and fenbendazole (FBZ) in cattle liver and muscle was collaboratively studied in 7 laboratories in 1986. For blind fortified samples containing 800 ppb FBZ, average recovery and relative standard deviations for repeatability and reproducibility (RSDr and RSDR) based on results from 6 of the participating laboratories were 83%, 12.7%, and 14.0%, respectively. Recoveries of FBZ from incurred liver samples were more variable. Recoveries of MBZ from livers fortified at the 100 ppb level were encouraging; however, the drug levels were too low in the incurred samples used for MBZ studies. Except for FBZ and MBZ in liver, the study data were not satisfactory. The method has been adopted official first action by AOAC for determination of 800-1600 ppb fenbendazole in liver. The analysis should be repeated using a smaller sample size when initial analyses show levels greater than 1600 ppb FBZ.     

Development of an analytical scheme for simazine and 2,4-D in soil and water runoff from ornamental plant nursery plots

The transport and fate of pesticides applied to ornamental plant nursery crops are not well documented. Methodology for analysis of soil and water runoff samples concomitantly containing the herbicides simazine (1-chloro-4,6-bis(ethylamino)-s-triazine) and 2,4-D ((2,4-dichlorophenoxy)acetic acid) was developed in this research to investigate the potential for runoff and leaching from ornamental nursery plots. Solid-phase extraction was used prior to analysis by gas chromatography and liquid chromatography. Chromatographic results were compared with determination by enzyme-linked immunoassay analysis. The significant analytical contributions of this research include (1) the development of a scheme using chromatographic mode sequencing for the fractionation of simazine and 2,4-D, (2) optimization of the homogeneous derivatization of 2,4-D using the methylating agent boron trifluoride in methanol as an alternative to in situ generation of diazomethane, and (3) the practical application of these techniques to field samples.     

Thin layer chromatographic detection and indirect gas chromatographic determination of three carbamate pesticides.

Carbamate pesticide residues are extracted from vegetables and fruits with methylene chloride. The extracts are spotted on silica gel plates and the pesticides are detected by an enzymatic inhibition technique. For quantitative determination, aliquots of the methylene chloride extracts are evaporated to dryness in a rotary evaporator. After the residues are dissolved in ethanol, 0.5N NaOH is added in the hydrolysis step. To remove a number of possible interferences the hydrolyzed phenols are steam-distilled and treated with 1-fluoro-2,4-dinitrobenzene and/or 4-chloro-alpha,alpha,alpha-trifluoro-3,5-dinitrotoluene to form the ether derivatives. Efficiency in the conversion of the phenolic moieties to the phenyl ethers is about 100%. The resulting electron-capturing derivatives enable the carbamate pesticides to be detected in vegetables and fruits at the 0.05 ppm level. Recoveries of 90-94% were obtained from vegetables and fruits fortified with 0.5-2.0 ppm carbaryl, Mesurol, and propoxur.     

Gas chromatographic method for determination of chlorpyrifos and its metabolite 3,5,6-trichloro-2-pyridinol (TCP) in dates.

A method is described for the determination of the insecticide chlorpyrifos and its metabolite TCP in green, unprocessed, and processed dates with the seeds incorporated. After extraction, chloropyrifos is cleaned up using Florisil and analyzed using a gas chromatography (GC) equipped with a nitrogen/phosphorus detector. TCP is derivatized using bis-(trimethylsilyl)-acetamide (BSA) to form the TCP-derivative and analyzed by a gas chromatograph equipped with a Hall electrolytic conductivity detector. Recoveries of chlorpyrifos from all fortified dates (0.05 and 0.1 ppm) ranged from 86 to 110% with an average of 94.5%. Recoveries of TCP from all fortified dates (0.1 and 0.2 ppm) ranged from 79 to 99% with an average of 86%. Limits of detection for chlorpyrifos and TCP in green, unprocessed, and processed dates were 0.02 and 0.05 ppm, respectively.     

Gas-liquid chromatographic determination of kepone residues in finfish, shellfish, and crustaceans.

Finfish, shellfish, and crustacean samples are extracted with isopropanol and benzene; the extract is filtered and then concentrated. The extract, dissolved in hexane, is treated with oleum and extracted with aqueous alkali. The aqueous phase is acidified and extracted with petroleum ether-ethyl ether (1 + 1). The Kepone residue is determined by electron capture gas-liquid chromatography (GLC). Recoveries obtained by 8 laboratories from 15 species of finfish fortified at 0.02-0.23 ppm ranged from 37 to 107% with a mean +/- relative standard deviation of 79.4 +/- 14.5%. For oysters fortified at 0.01-0.10 ppm, recoveries range from 63 to 129% with a mean of 78.8 +/- 20.8%. For crustaceans fortified at 0.05-0.26 ppm, recoveries ranged from 52 to 110% with a mean of 78 +/- 16.4%. The approximate limits of quantitation for finfish and for shellfish and crustaceans are 0.02 and 0.05 ppm, respectively, under the GLC conditions used in this study.     

Mass spectrometric identification and gas-liquid chromatographic determination of 2-chloroethyl esters of fatty acids in spices and foods.

The 2-chloroethyl esters of 5 fatty acids have been identified in spice and food samples by gas-liquid chromatography-mass spectrometry (GLC/MS). Twenty-four spice samples were analyzed for the 2-chloroethyl esters of fatty acids by AOAC official multiple residues pesticide procedure using GLC with microcoulometric detection. The esters of capric, lauric, myristic, palmitic, and linoleic acids have been identified at levels up to 1400 ppm. 2-Chloroethyl linoleate was the most abundant ester in all samples. Several foods analyzed by the same procedures showed levels of 2-chloroethyl linoleate as high as 35 ppm. Recoveries from fortified samples ranged from 84 to 98% for the various esters. A method using an acid-catalyzed esterification reaction was developed to rapidly determine the fatty acid content of these spices. GLC analysis with microcoulometric detection was used. Recoveries from fortified samples ranged from 92 to 110%. After 2 spice samples found to be free of 2-chloroethyl esters were fumigated with ethylene oxide, the level of 2-chloroethyl linoleate reached 77 ppm. All levels of 2-chloroethyl esters were confirmed by GLC/MS.     

Liquid chromatographic determination of paraquat and diquat in crops using a silica column with aqueous ionic mobile phase.

A method was developed for the determination of paraquat (PQ) and diquat (DQ) in high moisture food crops. Samples were digested with 6M HCl, and the herbicides were isolated from the digest using pH-controlled silica solid phase extraction. The analytes were then determined by ion-pairing liquid chromatography with a silica analytical column, sodium chloride as the ion-pairing reagent, and acetonitrile as the organic modifier. A diode array UV absorbance detector was used to simultaneously quantify PQ and DQ at their respective maximum absorbance wavelengths, 257 and 310 nm. Average recoveries of PQ and DQ standards from 4 different crops fortified at 0.01-0.50 ppm levels ranged from 79.3 to 104.8%.     

Purge and trap method for determination of fumigants in whole grains, milled grain products, and intermediate grain-based foods

A method developed for the determination of 1,2-dibromoethane in whole grains and grain-based products has been modified and expanded to include 8 other fumigants. Samples are stirred with water and purged with nitrogen for 0.5 h in a water bath at 100 degrees C. The fumigants are collected on a trap composed of Tenax TA and XAD-4 resin, eluted with hexane, and determined by gas chromatography (GC) using electron capture detection or Hall electrolytic conductivity detection. Flame photometric detection in the sulfur mode is used to determine carbon disulfide. Thick-film, wide-bore capillary columns were used exclusively in both the determination and confirmation of the halogenated fumigants. The higher levels of fumigants are also confirmed by full scan GC/mass spectrometry. Samples are analyzed for carbon disulfide, methylene chloride, chloroform, 1,2-dichloroethane, methyl chloroform, carbon tetrachloride, trichloroethylene, 1,2-dibromoethane, and tetrachloroethylene. A total of 25 whole grains, milled grain products, and intermediate grain-based foods analyzed by this method contained fumigant levels up to 51 ppm (carbon tetrachloride in wheat). Recoveries from fortified samples ranged from 82 to 104%. Chromatograms from this purge and trap method are clean, so that low parts per billion and sub-parts per billion levels can be quantitated for the halogenated analytes. The quantitation level for carbon disulfide is 12 ppb.