Gas-liquid chromatographic analysis and chemical confirmation of azodrin (monocrotophos) residues in strawberries.

A gas-liquid chromatographic (GLC) method is described for the analysis and confirmation of azodrin (monocrotophos, 3-(dimethoxyphosphinyl)-N-methyl-cis-crotonamide) residues in strawberrires. The strawberries are extracted with acetone, and the filtrate is partioned with a mixture of methylene chloride and petroleum ether followed by further extraction with methylene chloride. The organic phases are combined, dried with anhydrous sodium sulfate, and concentrated to a small volume for GLC analysis on a 3% OV-210 column with flame photometric detection. Identity of the compound is confirmed by chromatography on the same column after trifluoroacetylation of an aliquot of the strawberry extract. The detection limit is about 2 ppb. The types of strawberry samples analyzed were fresh, frozen, pureed, and jam.     

Determination of N-methylcarbamate pesticides in liver by liquid chromatography

A liquid chromatographic (LC) method using a 2-step purification technique for the simultaneous determination of 10 carbamates in bovine, swine, and duck livers has been developed. Carbamates are extracted from liver samples with methylene chloride. After evaporation, the residues from the extract are dissolved in methylene chloride-cyclohexane (1 + 1) and cleaned up by gel permeation chromatography. The eluate containing carbamate residues is evaporated to dryness, reconstituted in methylene chloride, further purified by passing it through an aminopropyl Bond Elut extraction cartridge, and analyzed by liquid chromatography using post-column derivatization with orthophthalaldehyde and fluorescence detection. Excitation and emission are set at 340 and 418 nm, respectively. Liver samples for beef, pork, and duck were fortified with 5, 10, and 20 ppb of mixed carbamate standards. The average of 10 recoveries of 10 carbamates at all 3 levels of fortification was greater than 80% with coefficients of variation less than 17%.     

Gas-liquid chromatographic determination of nikethamide in injectable preparations.

A gas-liquid chromatographic (GLC) method, using a 4% XE-60 on 80-100 mesh Gas-Chrom Q column, a flame ionization detector, and anthracene as the internal standard, has been developed for the direct determination of nikethamide. Eight collaborators analyzed 4 samples, using methanol as the solvent; the coefficients of variation obtained ranged from 1.19 to 3.20%. In a limited study with acetone as the solvent, the coefficients of variation ranged from 0.59 to 1.96%. The GLC method with acetone as a solvent has been adopted as official first action.     

Application of gel permeation chromatography and nonaqueous reverse phase chromatography to high pressure liquid chromatographic determination of retinyl palmitate and beta-carotene in oil and margarine.

A high pressure liquid chromatographic method was developed using high pressure gel permeation chromatography (HP-GPC) and high pressure reverse phase chromatography (RP-HPLC) for quantitation of retinyl palmitate and beta-carotene. HP-GPC was used for fractionation of vitamin A active compounds from oil preliminary to quantitation on nonaqueous RP-HPLC. HP-GPC fractionation was completed on oil and margarine dissolved in methylene chloride by 2 elution passes through 2 muStyragel (100 angstrom) columns connected in series with methylene chloride as the mobile phase. RP-HPLC separation of retinyl palmitate and beta-carotene was achieved on muBondapak C18 (10 micrometers), using methylene chloride-acetonitrile (30+70). Based on 10 repetitive analyses, recoveries of added beta-carotene and retinyl palmitate from vegetable oils were 98.6+/-2.9 and 95.2+/-2.6%, respectively. The coefficients of variation were 2.9% for beta-carotene and 2.7% for retinyl palmitate. The determination of vitamin A activity in 7 margarine brands with label claims of 10% U.S.RDA/serving revealed that all but one of the margarines contained at least 94% of the label claim. Vitamin A activity in the margarines ranged from 90.6 to 110.8% of the label declaration.     

Rapid method for determination of leucogentian violet in chicken fat by liquid chromatography with electrochemical detection

The metabolite leucogentian violet (LGV) was found in chicken fat obtained from chickens dosed with gentian violet (GV); however, no residues of the parent compound, GV, and its oxidized metabolites were found. Therefore, a rapid method was developed for the specific determination of LGV in chicken fat. Chicken fat containing LGV is separated from the cellular protein with methylene chloride. LGV is then separated from the fat by partition extraction with an aqueous acid phase in which LGV is protonated, and the fat is discarded with the methylene chloride layer. The aqueous solution is neutralized, LGV is re-extracted into methylene chloride, and the methylene chloride is evaporated. An acetonitrile-water solution containing LGV is filtered before liquid chromatography using a cyano column, an acetate buffer-acetonitrile mobile phase, and an electrochemical detector set at a potential of +1.000 V. Average recoveries of LGV from chicken fat were 83.9% with a coefficient of variation (CV) of 12.9% for the 5 ppb level; 82.8% with a CV of 13.5% for the 10 ppb level; and 77.7% with a CV of 2.56% for the 20 ppb level. Levels of incurred LGV in chicken fat averaged 49.3 ppb with a CV of 2.43%.     

Enzymatic determination of cholesterol in egg yolk

The quantitative determination of cholesterol in egg yolk by using an enzymatic test kit is described. Cholesterol in the egg yolk is extracted with other lipid components by methylene chloride-methanol (2 + 1) and is enzymatically determined after saponification of the lipid extract. The method is relatively rapid, simple, and accurate and gives results which agree with those obtained by using a gas-liquid chromatographic (GLC) method. The mean cholesterol content of egg yolk determined by the enzymatic and GLC methods was 1237 and 1240 mg/100 g, respectively.     

Determination of halogenated contaminants in human adipose tissue

A method has been developed for determination of organochlorine contaminants in human adipose tissue. After fat extraction from the tissue with acetone-hexane (15 + 85, v/v), organochlorines were fractionated from fat by gel permeation chromatography with methylene chloride-cyclohexane (1 + 1, v/v) as solvent. After Florisil column cleanup, the GPC extract was analyzed by capillary column gas chromatography using 2 columns of different polarity. Compound identity was confirmed by gas chromatography-mass spectrometry using selected ion monitoring. Recoveries for fortification levels of 10-500 ng/g were greater than 80% except for trichlorobenzene and hexachlorobutadiene (ca 60%).     

Determination of hexachlorobenzene and mirex in fatty products.

A procedure is described for the isolation and cleanup of hexachlorobenzene (HCB) and mirex in fats and oils for gas-liquid chromatographic (GLC) analysis. The fat or oil is distributed on unactivated Florisil, and the HCB and mirex are eluted with acetonitrile. The pesticides are then partitioned into petroleum ether. Elution through activated Florisil with methylene chloride-hexane (20+80) is used for the final cleanup. HCB and mirex are then measured by GLC, using the appropriate electron capture conditions with a 15% OV-210 column for HCB and a 3% OV-101 column for mirex. The method demonstrates recoveries greater than 90% for HCB and mirex and allows screening at or below the 0.1 ppm level in fats with a 3 mg fat injection.     

Extraction and cleanup of organochlorine, organophosphate, organonitrogen, and hydrocarbon pesticides in produce for determination by gas-liquid chromatography.

A rapid, multiresidue procedure utilizing the minimal cleanup necessary for gas-liquid chromatographic (GLC) analysis is presented. The samples are extraced with acetone and partitioned with methylene chloride-petroleum either to remove water. The organophosphorus and organonitrogen compounds are then quantitated by GLC, using a KCl thermionic detector. A Florisil cleanup of the extract is performed prior to the determination of organochlorine compounds by a GLC electron capture detector. Carbon-hydrogen compounds such as biphenyl and o-phenylphenol undergo the Florisil cleanup and may also be quantitated by GLC. Quantitative recoveries for 15 organophosphorus, 9 organochlorine, 5 organonitrogen, and 2 hydrocarbon pesticides show the range in polarities of pesticides recovered, from Monitor to biphenyl. The method is simple and fast with a great potential for the analysis of many more compounds.     

Determination of methylene chloride, ethylene dichloride, and trichloroethylene as solvent residues in spice oleoresins, using vacuum distillation and electron capture gas chromatography.

A quantitative gas chromatographic (GC) method is described for the determination of residual methylene chloride, ethylene dichloride, and trichloroethylene in spice oleoresins. The proposed method involves vacuum distillation in a closed system with toluene as a carrier solvent. Quantitation by electron capture GC on Porapak Q is facilitated by water extraction and by the addition of trans-1,2-dichloroethylene as an internal standard. Recoveries from oleoresins spiked at 30, 15, and 6 ppm ranged from 93 to 102%. To assess the possibility of interference from spice volatiles, the procedure was applied to 17 different spice oleoresins from 3 different manufacturers. No interferences were found, but methylene chloride levels up to 83 ppm and ethylene dichloride levels up to 23 ppm were detected. Trichloroethylene was not detected in any of the oleoresins.     

Isolation and liquid chromatographic determination of the cyclic peptide mycotoxin cyclosporin A from rice

A simple method for determination and quantitation of a cyclic peptide mycotoxin, cyclosporin A, in rice is presented. Rice inoculated with Trichoderma polysporum (Link ex Pers.) was extracted with methylene chloride after 4 weeks of incubation. Cyclosporin A was isolated from extracts by using open bed gel filtration column chromatography (LH-20, acetonitrile) and monitored with thin layer chromatography and reverse phase liquid chromatography coelution with a standard. Preliminary thin layer chromatographic methods were developed. Cyclosporin A was detected by iodine and after partial acid hydrolysis by ninhydrin and UV light. A liquid chromatographic method was developed that used a reverse phase disposable cartridge cleanup and isocratic chromatography with a reverse phase octadecylsilica column and a UV detector set at 212 nm. Recovery of cyclosporin A from spiked rice samples (mg/g range) was 85%.     

Mycotoxins in grain dust: method for analysis of aflatoxins, ochratoxin A, zearalenone, vomitoxin, and secalonic acid D

A multimycotoxin method is presented to quantitate aflatoxins, ochratoxin A, zearalenone, secalonic acid D, and vomitoxin in grain dust. Dust spiked with these mycotoxins was extracted sequentially with methylene chloride followed by acetonitrile-water (86 + 14). Vomitoxin was recovered in the latter extract and all other mycotoxins were recovered in the methylene chloride. Aflatoxins and ochratoxin were quantitated by fluorescence measurement on silica thin layer chromatographic plates. The other mycotoxins were quantitated after cleanup by reverse phase liquid chromatography and ultraviolet detection. Recoveries from dust spiked in the parts per billion (ng/g) range were approximately 80% (SD = 15-29%) for all mycotoxins. Minimum detectable amounts ranged from less than 0.5 ng/g for aflatoxins to 20 ng/g for zearalenone.     

Determination of polysorbates in foods by colorimetry with confirmation by infrared spectrophotometry, thin-layer chromatography, and gas chromatography

A method is presented for the detection of polysorbates (PSs) in 8 kinds of processed foods by colorimetric and thin-layer chromatographic (TLC) techniques. The PSs are extracted from processed foods with a mixture of methylene chloride and ethanol by using an Extrelut column. The extract is further purified by using a silica gel column. The PS extract is complexed with cobalt-thiocyanate (Cothiocyanate) reagent and is determined spectrophotometrically at 620 nm. The recoveries and coefficients of variation for 8 kinds of processed foods fortified with 0.1% PS 80 were 67.9-94.6% and 4.0-11.3%, respectively. The detection limit of TLC corresponded to 50 mg PS 80/kg. PS identity was confirmed by infrared spectrophotometry of PS extract, and gas chromatography of fatty acids and thin layer chromatography of POE-sorbitan residues after saponification.     

Gas-liquid chromatographic determination of monosodium glutamate in soups and soup bases.

A method is given for determining monosodium glutamate (MSG) in soups and soup bases by gas-liquid chromatography (GLC) of the trimethylsilyl ether derivative of glutamic acid. This method compared favorably with existing methods including GLC of the trifluoroacetate (TFA)/butyl ester derivative, and analysis on an amino acid analyzer. In addition, some samples were analyzed by GLC/mass spectrometry. The MSG content of various products ranged from approximately 0.2% in some condensed soups to 13.1% on bouillon cubes. The method, which can detect as little as 0.05%, requires only a 10 min single step derivatization at room temperature and is preferred to the TFA/butyl ester technique.     

A routine high-performance liquid chromatography method for carotenoid determination in ultrafrozen orange juices

An isocratic reversed-phase high-performance liquid chromatography method was developed for routine analysis of the main carotenoids related to the color of orange juice, using a more selective wavelength (486 nm) in which the absorption in the red-orange region of the visible spectra is maximum. Separation was carried out using as the mobile phase the mixture methanol:acetonitrile:methylene chloride:water (50:30:15:5, v/v/v/v), to which small amounts of butylated hydroxytoluene and triethylamine were added (0.1%). Identification was made by comparison either with standards obtained by thin-layer chromatography or with spectral data previously reported. The reproducibility of the method was remarkable; coefficients of variation for the most polar xanthophylls were under 1 and 4% for retention times and areas, respectively. Its application to Valencia late ultrafrozen orange juices has shown that major carotenoids are lutein + zeaxanthin (36%), lutein 5,6-epoxide (16%), antheraxanthin (14%), and beta-cryptoxanthin (12%).     

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%.     

A Comparison of Spectrophotometric and Gas Chromatographic Measurements of Heavy Petroleum Products in Soil Samples

Laboratory studies were conducted to compare the infrared spectrophotometry (TPH-IR) and gas chromatography (TPH-GC) measurements of total petroleum hydrocarbon in soil samples.Real world soil samples containing #2 to #6 fuel oils, mechanical lubricating oil, diesel fuel, kerosene, jet fuel andweathered gasoline were extracted with trichlorotrifluoroethane(Freon-113) and methylene chloride. The extractants were analyzed using gas chromatography with flame ionization detection (GC-FID) and infrared spectroscopy (TPH-IR) methods.A paired statistical t-test was applied to compare the average of paired differences in the analytical results. Statistical tests were evaluated with graphical presentation of the results. In general, a trend was observed in the measured concentrations.Total petroleum hydrocarbon (TPH) concentrations measured withTPH-IR had the highest readings. The same samples extracted withmethylene chloride and analyzed with GC-FID showed lower concentrations than the TPH-IR method while the GC-FID analysisof the same samples extracted with Freon-113 produced the lowestconcentrations. Laboratory experiments indicated that TPH concentrations measured with the TPH-IR method were higher thanthe actual quantities of petroleum hydrocarbon in the soil samples.     

Determination of polybrominated biphenyl residues in dairy products.

A method for the determination of polybrominated biphenyls (PBBs) in dairy products is described. Fat is extracted from the products by the official AOAC method. The PBB residues are separated from the fatty material by gel permeation chromatography prior to gas-liquid chromatographic (GLC) quantitation. An additional cleanup using petroleum ether elution through a miniature Florisil column is necessary for thin layer chromatographic (TLC) confirmation. Recoveries of PBBs from samples fortified at levels from 0.1 to 0.5 ppm ranged from 94 to 104% with an average of 99%. GLC sensitivity permits the estimation of PBB residue levels as low as 0.007 ppm. Routine TLC confirmation is limited by sensitivity to greater than or equal to 0.2 ppm.     

Gas-liquid chromatographic determination of beta-asarone in wines and flavors.

Wine samples containing beta-asarone (cis-2,4,5-trimethoxy-l-propenylbenzene) are distilled; beta-asarone is extracted by hexane and then quantitatively determined by gas-liquid chromatography (GLC), using ethyl palmitate as the internal standard. The GLC procedure is rapid and yields precise and accurate results. Mass spectrometry confirmed the identity of the GLC peak as beta-asarone. The ultraviolet spectra of beta-asarone and its isomer were also determined.     

Determination of altosid insect growth regulator in waters, soils, plants, and animals by gas-liquid chromatography.

Residues of isopropyl (2E,4E)-11-methoxy-3,7,11-trimethyl-2,4-dodecadienoate (Altosid) insect growth regulator are determined in waters, soils, plants, milk, eggs, fish, shellfish, poultry and cattle tissues, blood, urine, and feces. Acetonitrile is the primary extraction solvent for all samples. Residues are extracted by high-speed blending followed by vacuum filtration. Fatty extracts are subjected to cold-temperature precipitation and filtration. Samples are cleaned up by petroleum ether partitioning and Florisil and neutral alumina chromatography. The concentrated eluants are analyzed by gas-liquid chromatography (GLC) on columns of differing polarity, using hydrogen flame ionization detectors. The identity of suspected residues is confirmed by additional GLC and by mass fragmentography. The lower limits of detection were: water samples, 0.0004-0.001 ppm; soils, blood, and urine, 0.001 ppm; forage grasses, forage legumes, and rice foliage, 0.005 ppm; and milk, eggs, fish, shellfish, poultry and cattle tissues, and feces, 0.010 ppm.