Improved cleanup and derivatization for gas chromatographic determination of monosodium glutamate in foods

A gas chromatographic procedure is described for determining monosodium glutamate (MSG) in several types of food. A sample is extracted with acetone-water (1 + 1). Acetone is evaporated and an aliquot of the extract is buffered with 1M NH4OH-1M NH4Cl pH 9 solution, and chromatographed directly on a column of QAE Sephadex A-25 that has been pretreated with the same buffer. MSG is eluted with 0.1N HCl, and a portion of the eluate is evaporated to dryness and reacted with dimethylformamide(DMF)-dimethylacetal to form the glutamic acid derivative, which is injected into a gas chromatograph and measured by flame ionization detection. Recoveries of MSG from sample fortified at 5-500 mg ranged from 92.8 to 100%.     

Use of solid-phase extraction systems to improve the sensitivity of Artemia bioassays for trichothecene mycotoxins

Solid-phase extraction was used to preconcentrate trichothecene mycotoxins from rivers and streams in order to develop and improve a rapid and sensitive bioassay using the brine shrimp Artemia salina. For T-2 toxin, HT-2 toxin, and 4,15-diacetoxyscirpenol, LC50 values obtained were 172, 600, and 700 micrograms/L, respectively. The LC50 for 4-deoxynivalenol was 21 mg/L. A more than 5-fold increase in sensitivity was observed when solid-phase extraction (SPE) was used in conjunction with the Artemia bioassay. For T-2 toxin, HT-2 toxin, and 4,15-diacetoxyscirpenol, LC50/SPE values after solid-phase extraction were 21, 83, and 130 micrograms/L. The use of river and stream waters and chlorinated water did not seem to interfere with the bioassay.     

Aflatoxins in domestic and imported foods and feeds

Aflatoxins, metabolic products of the molds Aspergillus flavus and A. parasiticus, may occur in foods and feeds. These toxins cannot be entirely avoided or eliminated from foods or feeds by current agronomic and manufacturing processes and are considered unavoidable contaminants. To limit aflatoxin exposure, the U.S. Food and Drug Administration (FDA) has set action levels for these toxins in foods and feeds involved in interstate commerce. FDA continually monitors food and feed industries through compliance programs. This report summarizes data generated from compliance programs on aflatoxins for the fiscal year 1986. Commodities sampled included peanuts and peanut products, corn and corn products, tree nuts, cottonseed, milk, spices, manufactured products, and miscellaneous foods and feeds. Correlations were highest between aflatoxin contamination and geographical areas for corn/corn products and cottonseed/cottonseed meal. Higher incidences of aflatoxin contamination in corn and corn products designated for human consumption were observed in samples collected in the southeastern states (32 and 28%, respectively). A higher incidence of contamination was observed in corn designated for animal feed from Arkansas-Texas (74%) than from the southeastern states (47%). Only 3% of feed corn from corn belt states contained detectable aflatoxins. All aflatoxin-contaminated cottonseed was collected in the Arizona-California area; 80% of cottonseed meal analyzed from this area also contained detectable levels of aflatoxins.(ABSTRACT TRUNCATED AT 250 WORDS)     

Worldwide occurrence of mycotoxins in foods and feeds--an update

In a review presented at the first FAO/WHO/UNEP Conference on Mycotoxins in 1977, the occurrence of aflatoxins, zearalenone, ochratoxin A, citrinin, trichothecenes, patulin, penicillic acid, and the ergot alkaloids was indicated to be significant in naturally contaminated foods and feeds. The information presented on aflatoxin contamination greatly exceeded that for all other mycotoxins combined. This study reviews the worldwide levels and occurrence of mycotoxins in various commodities since 1976. Comparatively few countries have lowered the acceptable levels for aflatoxins in susceptible commodities. However, intensified efforts are needed to establish control of aflatoxin levels in the global food supply, particularly in peanuts, tree nuts, corn, and animal feeds. Extensive deoxynivalenol (DON) contamination of grains, especially wheat, was demonstrated. Co-contamination of grains by Fusarium toxins, especially DON and nivalenol, with zearalenone to a lesser extent, was reported. However, more information on co-occurrence of Fusarium toxins in cereals should be developed. When contamination of feeds by ochratoxin A was significant, this toxin occurred in swine kidney and smoked meats in high levels. On the basis of occurrence and/or toxicity, patulin and penicillic acid contamination of foods does not appear to be of real concern. More recent developments suggest, however, that expanded monitoring studies of Alternaria toxins, moniliformin, citrinin, cyclopiazonic acid, penitrem A, and ergot alkaloids are indicated.     

Use of ten gram samples of corn for determination of mycotoxins

Data were gathered, during a study on the development of an automated system for the extraction, cleanup, and quantitation of mycotoxins in corn, to determine if it was scientifically sound to reduce the analytical sample size. Five, 10, and 25 g test portions were analyzed and statistically compared with 50 g test portions of the same composites for aflatoxin concentration variance. Statistical tests used to determine whether the 10 and 50 g sample sizes differed significantly showed a satisfactory observed variance ratio (Fobs) of 2.03 for computations of pooled standard deviations; paired t-test values of 0.952, 1.43, and 0.224 were computed for each of the 3 study samples. The results meet acceptable limits, since each sample's t-test result is less than the published value of the /t/, which is 1.6909 for the test conditions. The null hypothesis is retained since the sample sizes do not give significantly different values for the mean analyte concentration. The percent coefficients of variation (CVs) for all samples tested were within the expected range. In addition, the variance due to sample mixing was evaluated using radioisotope-labeled materials, yielding an acceptable CV of 22.2%. The variance due to the assay procedure was also evaluated and showed an aflatoxin B, recovery of 78.9% and a CV of 11.4%. Results support the original premise that a sufficiently ground and blended sample would produce an analyte variance for a 10 g sample that was statistically comparable with that for a 50 g sample.     

Liquid chromatographic determination of sulfamethazine in feeds

A reverse-phase liquid chromatographic method for the assay of sulfamethazine (SMZ) in feeds is described. Feed samples are extracted with 50% methanol solution, centrifuged, filtered, and diluted when necessary, and chromatographed on a C-18 column. Samples are eluted with a mobile phase of 20% methanol and 80% of a solution containing acetic acid and tetramethylammonium chloride. The average recovery from spiked samples was 97.2% with a coefficient of variation of 1.2%. Linearity was very good (correlation coefficient 0.9997). Within-day and between-day coefficients of variation averaged 1.3 and 2.6%, respectively. The results for samples assayed by this method compared closely with the results from the same extracts assayed by the AOAC colorimetric method.     

Gas chromatographic determination of electron capture sensitive volatile industrial chemical residues in foods, using AOAC pesticide multiresidue extraction and cleanup procedures

Electron capture (EC) gas chromatographic (GC) parameters have been developed for determining some of the more volatile industrial chemicals that can be determined by the AOAC multiresidue method for organochlorine and organophosphorus pesticides with modified GC operating conditions. Retention times relative to pentachlorobenzene are reported for 143 industrial chemicals, pesticides, and related compounds on OV-101 GC columns at 130 degrees C. Also reported for most of the compounds are recoveries from fortified samples carried through the AOAC extraction and cleanup procedures for fatty and/or nonfatty foods, Florisil elution characteristics, and GC relative retention times on mixed OV-101 + OV-210 columns at 130 degrees C. Our laboratory has used the modified EC/GC parameters with the AOAC multiresidue extraction/cleanup procedures to determine many volatile halogenated industrial chemical contaminants in foods, chiefly in samples of fresh-water fish. Other modifications of the AOAC method are described to improve the tentative identification and quantitative measurement of these volatile residues.     

Liquid chromatographic determination of aflatoxins, ochratoxin A, and zearalenone in grains, oilseeds, and animal feeds by post-column derivatization and on-line sample cleanup

A liquid chromatographic method using on-line sample cleanup, reverse flow analytical column loading, gradient elution, and postcolumn derivatization with iodine permits direct, rapid determination of aflatoxins B1, B2, G1, and G2, as well as ochratoxin A and zearalenone. Limits of quantitation are 5 ppb for the aflatoxins and ochratoxin A and 30 ppb for zearalenone. This procedure performs well as a multimycotoxin screen for cereal grains and oilseeds, with more limited success in complete animal feeds.     

Use of the Mycosep multifunctional cleanup column for liquid chromatographic determination of aflatoxins in agricultural products.

A liquid chromatographic (LC) technique has been developed that uses the Mycosep multifunctional cleanup (MFC) column. MFC columns provide a rapid 1-step extract purification. They are designed to retain particular groups of compounds that may create interferences in analytical methods. At the same time, MFC columns allow compounds of interest to pass through. In the method presented, test samples are extracted in a blender with acetonitrile-water (9 + 1). A portion of the extract is forced through an MFC column designed especially for analysis of numerous mycotoxins. Analytical interferences are retained, while aflatoxins pass through the column. Aflatoxins B1 and G1 are converted to their hemiacetals by heating a mixture of purified extract and water-trifluoroacetic acid-acetic acid (7 + 2 + 1) at 65 degrees C for 8.5 min. An aliquot of this mixture is analyzed by isocratic LC with acetonitrile-water mobile phase and fluorescence detection. A detection limit of less than 0.5 ng/g for aflatoxin B1 was obtained. Average recoveries greater than 95% total aflatoxins (B1, B2, G1, and G2) and coefficients of variation of less than 3% were obtained. The method was successfully applied to the following commodities: corn, almonds, pista-chios, walnuts, peanuts, Brazil nuts, milo, rice, cottonseed, corn meal, corn gluten meal, fig paste, and mixed feeds.     

Liquid chromatographic determination of chlortetracycline hydrochloride in ruminant and poultry/swine feeds.

A liquid chromatographic (LC) method is described for the determination of chlortetracycline hydrochloride (CTC) in poultry/swine and ruminant feeds in the 10-100 ppm range and in premix. CTC is extracted from ground feed/premix with acidified acetone, and the extract is filtered through a Millex-HV filter or disposable C18 column. The filtrate is partitioned with methylene chloride when additional cleanup is necessary. A Nova-Pak C18 column is used for LC separation with determination at 370 nm. The average recovery of CTC from premix was 95% with a standard deviation (SD) of 1.70 and a coefficient of variation (CV) of 1.79%. The overall average recovery from feeds was 77% with an SD of 3.18 and a CV of 4.10%.     

Microbiological determination of neomycin in feeds and formulated products

An AOAC modified method is described for the microbiological assay of neomycin, which has been adapted to include complete feeds, supplements, premixes, liquids, oil suspensions, boluses, and antibiotic-impregnated paper. The method features a more sensitive standard response line with a monolayer plating system. The use of a buffered plating medium in place of the water-prepared medium results in a curve with less degree of slope, which allows for more accurate interpretation of the standard response. The feed extract diluent used for standard response line dilution, which is prepared from exposure of the feed extract fluid to pH changes, heat, and sodium hypochlorite, has been eliminated. The constant salt concentration diluent used for the preparation of standards is the same as the salt concentration of the sample extract solution to be tested. Results for 50 commercial complete feeds and 50 commercial premixes received over the last 5 years produced an overall mean recovery of 101% with a mean percent recovery range of 80-112%. A statistical analysis of these 100 commercial, complete feeds and premixes, ranging in concentration from 47 g/ton to 70 g/lb, indicates the assay has little, if any, concentration-related bias. Precision and accuracy of the method was supported by laboratory studies of 20 assays that produced a mean recovery of 101% and standard deviation of 3.     

Liquid chromatographic determination of oxfendazole in swine feeds

A relatively simple analytical method is presented for determination of oxfendazole (2-(methoxycarbonylamino)-5-phenylsulfinyl-benzimidazole) at levels as low as 0.012% in swine feeds, using cation exchange liquid chromatography (LC). The sample was extracted with a solvent mixture of methanol-glacial acetic acid (90 + 10) at 45 degrees C, using a gyrorotory shaker. Plant pigments and other feed excipients were removed using zinc acetate treatment and pH-controlled extraction. Oxfendazole was further separated from the remaining interferences and quantitatively determined by LC on a Partisil SCX column with acetonitrile-0.01M phosphate buffer as mobile phase. The method is stability-specific, linear, precise, and accurate at 80-120% labeled strength (relative standard deviation 0.9-1.7 with mean recovery of 98-99%). Supporting data at a level of 0.0135% oxfendazole in swine feed indicated that this method is capable of complete recovery of oxfendazole from medicated swine feeds.     

Simplified cleanup and gas chromatographic determination of organophosphorus pesticides in crops

A simple and efficient cleanup method for gas chromatographic determination of 23 organophosphorus pesticides in crops including onion is described. The sample was extracted with acetone. The extract was purified with coagulating solution, which contained ammonium chloride and phosphoric acid, and then filtered by suction. The filtrate was diluted with NaCl solution and reextracted with benzene. The organic layer was evaporated and injected into a gas chromatograph equipped with a flame photometric detector (FPD) and fused silica capillary columns (0.53 mm id) coated with silicone equivalent to OV-1701, OV-1, and SE-52. Onion extract, which contained FPD interferences, was cleaned up on a disposable silica cartridge. Recoveries of most organophosphorus pesticides from spiked crops: mandarin orange, tomato, spinach, sweet pepper, broccoli, lettuce, and onion at levels of 0.02-0.28 ppm, exceeded 80%, but the water-soluble pesticides dichlorvos and dimethoate gave poor recoveries in all crops; the nonpolar pesticides disulfoton, chlorpyrifos, fenthion, prothiophos, and leptophos were not recovered quantitatively in spinach, sweet pepper, broccoli, and lettuce. IBP, edifenphos, phosmet, and pyridaphenthion were not recovered from onion because of adsorption to the silica cartridge. The detection limits ranged from 1.25 to 17.5 ppb on a crop basis.