Derivative method for chemical confirmation of identity of aflatoxin M.
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Acetals↗
Biomedical subjects
M E Stack
Publications and source records attributed to M E Stack.
Mutagenic perylenequinone metabolites of Alternaria alternata: altertoxins I, II, and III.
The mold genus Alternaria is a widely distributed plant pathogen. Some of these species, e.g., A. alternata, are common decay organisms of fruits and vegetables. Two novel perylene oxide metabolites, altertoxins II and III, have been identified in extracts of A. alternata isolates that exhibit mutagenic responses in the Ames Salmonella typhimurium assay. These identifications were based on mass, optical rotational, and 1H- and 13C-nmr spectral studies. Previous reports of related perylene dione mycotoxins have been clarified.
Alternaria↗
Stemphyltoxin III from Alternaria alternata.
Stemphyltoxin III, (6aR*, 6bS*, 7R*, 8R*)-3,6a, 10-trihydroxy-4,9-dioxo-4,6a,6b,7,8,9-hexahydro-7,8-epoxyperylene, a known metabolite of Stemphylium botryosum var. lactucum, has been identified as a mutagenic metabolite of Alternaria alternata by spectroscopic studies. The 13C-nmr spectral data, which were not reported previously, are presented.
Alternaria↗
Isolation and identification of dihydrocitrinone, a urinary metabolite of citrinin in rats.
Dihydrocitrinone, 3,4-dihydro-6,8-dihydroxy-3,4,5-trimethylisocoumarin-7-carboxylic acid, was isolated and identified as a urinary metabolite after oral administration of citrinin to rats. Male and female Osborne-Mendel rats received 30 mg citrinin/kg body weight by oral intubation. The metabolite dihydrocitrinone was present in urine collected at 0-2, 2-4, 4-6, 6-8, and 8-24 h after treatment. Only unchanged citrinin was found in blood collected 24 h after administration of the compound. The metabolite had a blue fluorescence and the same Rf on thin-layer chromatography, the same retention time on reverse-phase high-pressure liquid chromatography, and the same mass spectrum as an authentic sample of dihydrocitrinone.
Animals↗
Immunoaffinity column coupled with solution fluorometry or liquid chromatography postcolumn derivatization for determination of aflatoxins in corn, peanuts, and peanut butter: collaborative study.
An AOAC/IUPAC (International Union of Pure and Applied Chemistry) collaborative study was conducted to evaluate the effectiveness of an immunoaffinity column for the determination of aflatoxin. The test portion is extracted with methanol-water (7 + 3), filtered, diluted to less than 30% methanol with water, and applied to the affinity column. The column is washed with water and the concentrated aflatoxins are eluted with methanol. Total aflatoxins are determined by solution fluorometry with bromine (SFB), and individual toxins are determined by reverse-phase liquid chromatography with postcolumn derivatization with iodine (PCD). Corn naturally contaminated with aflatoxins, and peanuts, peanut butter, and corn containing added aflatoxins (B1:B2:G1:G2 = 7:1:3:1) were sent to 24 collaborators in the United States, France, Canada, and the Republic of South Africa. Twelve collaborators used the SFB method, 9 used the PCD method, and 3 used both SFB and PCD methods. Twenty collaborators completed the study (10 used the SFB method, 7 used the PCD method, and 3 used both SFB and PCD methods). Test portions were spiked at 10, 20, and 30 ng/g. For SFB analyses, recoveries of total aflatoxins were 123, 105, and 107%, respectively; the relative standard deviation for repeatability (RSDr) ranged from 11.75 to 16.57%, and the relative standard deviation for reproducibility (RSDR) ranged from 10.97 to 33.09%. For PCD analyses, recoveries were 81, 81, and 83%, respectively; the RSDr ranged from 5.20 to 17.22%, and the RSDR ranged from 4.68 to 50.77%. The RSDr for aflatoxins B1 and G1 for spiked test portions ranged from 5.45 to 23.55%, and the RSDR ranged from 4.21 to 57.28%.(ABSTRACT TRUNCATED AT 250 WORDS)
Aflatoxins↗
Liquid chromatographic method for determination of aflatoxins B1, B2, G1, and G2 in corn and peanut products: collaborative study.
A collaborative study of a liquid chromatographic method for the determination of aflatoxins B1, B2, G1, and G2 was conducted in laboratories located in the United States, Canada, South Africa, and Switzerland. Twenty-one artificially contaminated raw peanuts, peanut butter, and corn samples containing varying amounts of aflatoxins B1, B2, G1, and G2 were distributed to participating laboratories. The test portion was extracted with methanol-0.1N HCl (4 + 1), filtered, defatted with hexane, and then partitioned with methylene chloride. The concentrated extract was passed through a silica gel column. Aflatoxins B1 and G1 were derivatized with trifluoroacetic acid, and the individual aflatoxins were determined by reverse-phase liquid chromatography with fluorescence detection. Statistical analysis of the data was performed to determine or confirm outliers, and to compute repeatability and reproducibility of the method. For corn, relative standard deviations for repeatability (RSDr) for aflatoxin B1 ranged from 27.2 to 8.3% for contamination levels from 5 through 50 ng/g. For raw peanuts and peanut butter, RSDr values for aflatoxin B1 were 35.0 to 41.2% and 11.2 to 19.1%, respectively, for contamination levels from 5 through 25 ng/g. RSDr values for aflatoxins B2, G1, and G2 were similar. Relative standard deviations for reproducibility (RSDr) for aflatoxin B1 ranged from 15.8 to 38.4%, 24.4 to 33.4%, and 43.9 to 54.0% for corn, peanut butter, and raw peanuts, respectively. The method has been adopted official first action for the determination of aflatoxins B1, B2, G1, and G2 in peanut butter and corn at concentrations greater than or equal to 13 ng total aflatoxins/g.
Aflatoxin B1↗
Enzyme-linked immunosorbent assay of aflatoxins B1, B2, and G1 in corn, cottonseed, peanuts, peanut butter, and poultry feed: collaborative study.
A direct competitive enzyme-linked immunosorbent assay (ELISA) screening method for aflatoxins at 20 ng/g was studied by 12 collaborators. Test samples of peanut butter were extracted by blending with methanol-water-hexane (55 + 45 + 100) and heating the test extracts on a steam bath; test samples of the other commodities were extracted by blending with methanol-water (80 + 20). All test extracts were filtered and the filtrates were diluted with buffer to a final methanol concentration of less than 30%. Each diluted filtrate was applied to a cup containing a filter with immobilized polyclonal antibodies specific to aflatoxins B1, B2, and G1. Aflatoxin B1-peroxidase conjugate was added, the cup was washed with water, and a mixture of hydrogen peroxide and tetramethylbenzidine was added. The test sample was judged to contain greater than or equal to 20 ng aflatoxins/g when, after exactly 1 min, no color was observed on the filter; when a blue or gray color developed, the test sample was judged to contain less than 20 ng aflatoxins/g. All collaborators correctly identified naturally contaminated corn and raw peanut positive test samples. No false positives were found for controls containing less than 2 ng aflatoxins/g. The correct responses for positive test samples spiked at levels of 10, 20, and greater than or equal to 30 ng aflatoxins/g (the ratio of B1:B2:G1 was 10:1:3) were 52, 86, and 96%, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)
Aflatoxin B1↗
Liquid chromatographic determination of tenuazonic acid and alternariol methyl ether in tomatoes and tomato products.
A liquid chromatographic (LC) method for determining tenuazonic acid (TA) and alternariol methyl ether (AME) in tomatoes and tomato products is described. The Alternaria metabolites are extracted from a water slurry of the sample with CHCl3, the mixture is centrifuged, and the extract is fractionated on a silica gel column. Reverse phase LC with an ultraviolet detector (for TA) and a fluorescence detector (for AME) connected in series is used for final separation and determination. The limit of determination for TA and AME is 25 and 3 ng/g, respectively, with average recoveries from catsup of 83 and 68%, respectively. The LC method also detects alternariol, but interfering peaks in some samples prevent accurate quantitation. Chemical ionization mass spectrometry (CIMS) is used to confirm TA. Samples (142) of tomatoes collected from commercial processing lines were analyzed; TA was found in 73 samples (0.4-70 micrograms/g).
Alternaria↗
Survey of deoxynivalenol in U.S. 1993 wheat and barley crops by enzyme-linked immunosorbent assay.
Wheat and barley from the 1993 crop year were analyzed for deoxynivalenol (DON). A total of 630 samples were collected by the Federal Grain Inspection Service in 25 states and analyzed using a commercially available, direct competitive, enzyme-linked immunosorbent assay. The limit of determination was about 0.5 micrograms/g. DON contamination in the 483 wheat samples averaged 2.0 micrograms/g and ranged from < 0.5 to 18 micrograms/g. DON contamination in the 147 barley samples averaged 4.2 micrograms/g and ranged from < 0.5 to 26 micrograms/g. About 40% fo the wheat samples and 57% of the barley samples contained DON levels that were greater than the U.S. Food and Drug Administration 1982 advisory level of 2 micrograms/g for DON in wheat designated for milling (human consumption).
Enzyme-Linked Immunosorbent Assay↗
Immunoaffinity column coupled with liquid chromatography for determination of fumonisin B1 in canned and frozen sweet corn.
A modified liquid chromatographic (LC) method for determining fumonisin B1 (FB1) in corn was applied to canned and frozen sweet corn. The corn is extracted with methanol-water (8 + 2), and the extract is filtered. The filtrate is diluted with water and passed through an immunoaffinity column. After the column is washed with water, FB1 is eluted with methanol-water (8 + 2). The eluate is evaporated to dryness by using a vacuum concentrator, and the residue is dissolved in acetonitrile-water (1 + 1). FB1 is derivatized with o-phthaldialdehyde. The derivative is separated on a reversed-phase C18 LC column using acetonitrile-water-acetic acid (50 + 50 + 1) and quantitated with a fluorescence detector. Recoveries of FB1 from canned and frozen corn spiked over the range of 50-200 ng/g were 76-88%. The limit of determination was about 25 ng/g, and the limit of detection was about 4 ng/g. The method was applied to 97 commercial canned and frozen sweet corn samples collected from different areas of the United States. Sixty samples contained no FB1. Low levels (trace-82 ng FB1/g corn) were found in 35 samples; 235 ng FB1/g was found in 1 canned corn sample, and 350 ng FB1/g was found in 1 frozen corn sample.
Chromatography, Affinity↗
Multifunctional column coupled with liquid chromatography for determination of aflatoxins B1, B2, G1, and G2 in corn, almonds, brazil nuts, peanuts, and pistachio nuts: collaborative study.
An AOAC/IUPAC collaborative study was conducted to evaluate the effectiveness of a multifunctional column for the determination of aflatoxins. The test portion is extracted with acetonitrile-water (9 + 1), the extract is filtered, and the filtrate is passed through the column. The aflatoxins in the eluate are determined by reversed-phase liquid chromatography after derivatization with trifluoroacetic acid. Naturally contaminated corn, almonds, Brazil nuts, peanuts, and pistachio nuts spiked with total aflatoxins at 5, 10, 20, and 30 ng/g were sent to 12 collaborators in the United States, Denmark, France, Japan, and Switzerland. Eleven collaborators completed the study. Average recoveries of total aflatoxins for each spike level for the various commodities (excluding Brazil nuts at 5 ng/g) were 93, 97, 95, and 95%, respectively; the repeatability relative standard deviation (RSDr) ranged from 6.0 to 23.2% and the reproducibility relative standard deviation (RSDR) ranged from 12.0 to 69.4%. The multifunctional column coupled with a liquid chromatographic method for determination of aflatoxins in corn, almonds, Brazil nuts, peanuts, and pistachio nuts has been adopted first action by AOAC INTERNATIONAL.
Aflatoxins↗
Determination of deoxynivalenol in 1991 U.S. winter and spring wheat by high-performance thin-layer chromatography.
A thin-layer chromatographic (TLC) method was modified for determination of deoxynivalenol (DON) in 1991 U.S. winter and spring wheat. After extraction with acetonitrile-water (84 + 16) and cleanup on a charcoal-alumina-Celite (7 + 5 + 3) column, acetonitrile was used instead of ethyl acetate to transfer the concentrated extract containing DON. After the extract was evaporated to dryness, the residue was dissolved in methanol and an aliquot was spotted on a high-performance TLC plate. After development with chloroform-acetone-2-propanol (8 + 1 + 1), the plates were sprayed with aluminum chloride solution and heated; DON was quantitated by fluorodensitometry. Average recoveries of DON added to duplicate test portions of wheat at 200, 400, and 800 ng/g were 83, 82, and 72%, respectively. The detection limit was 40 ng/g. The method was applied to 81 test samples of spring and winter wheat. The wheat contained DON levels that ranged from nondetectable to 9330 ng/g (average 1570 ng/g). The results indicate that DON levels were higher in wheat from Missouri, North Dakota, and Tennessee than in wheat from 7 other states. The identity of DON, which was isolated from 21 of the extracts by preparatory TLC, was confirmed by gas chromatography/mass spectrometry in all 21 test samples.
Chromatography, High Pressure Liquid↗
Enzyme-linked immunosorbent assay of total aflatoxins B1, B2, and G1 in corn: follow-up collaborative study.
A direct competitive enzyme-linked immunosorbent assay screening method for aflatoxins at 20 ng/g in corn was studied by 15 collaborating laboratories. Test samples of corn were extracted by blending with methanol-water (8 + 2). The extracts were filtered and the filtrates were diluted with buffer to a final methanol concentration of < 30%. Each diluted filtrate was applied to a test device containing a filter with immobilized polyclonal antibodies specific to aflatoxins B1, B2, and G1. Aflatoxin B1-peroxidase conjugate was added, the test device was washed with water, and a mixture of hydrogen peroxide and tetramethylbenzidine was added. A test sample was judged to contain > or = 20 ng aflatoxins/g when, after exactly 1 min, no color was observed on the filter; if a blue or gray color developed, the test sample was judged to contain < 20 ng aflatoxins/g. All laboratories correctly identified naturally contaminated corn test samples. Only one false positive was found for controls containing no aflatoxins. The correct responses for positive test samples spiked at levels of 10, 20, and 30 ng aflatoxins/g (the ratio of B1:B2:G1 was 15:1:3) were 67, 97, and 100%, respectively. This method was adopted first action by AOAC INTERNATIONAL as a change in method for 990.34 for screening for aflatoxins B1, B2, and G1 in corn at total aflatoxin concentrations of > or = 20 ng/g.
Aflatoxin B1↗
Fumonisin levels in Uruguayan corn products.
A survey was conducted to evaluate fumonisins FB1 and FB2 in Uruguayan corn products. Sixty-four samples of different local brands were purchased from retail stores during a 15-month period and analyzed for FB1 and FB2 by methanol-water extraction, cleanup with a 1 mL. strong-anion-exchange solid-phase extraction column, and liquid chromatography with o-pthaldialdehyde-2-mercaptoethanol derivatization and fluorescence detection. Contamination levels for FB1 varied from 50 ng/g (detection limit) to 6342 ng/g. Values were highest in feed samples (up to 6342 ng/g), unprocessed corn kernel (up to 3688 ng/g), and milled products, which included polenta (up to 427 ng/g). They were lowest in processed corn kernel (up to 155 ng/g) and snacks (up to 314 ng/g). FB2 was determined in one-fourth of the total samples and detected at trace levels in only one feed sample. The data demonstrated the natural occurrence of fumonisins in corn products in Uruguay. Feed and polenta that contain fumonisins could be of concern because they are consumed in large amounts and are often the main nutrient source in Uruguay.
Anion Exchange Resins↗
Analysis of fumonisin B1 and its hydrolysis product in tortillas.
Fumonisins are toxic metabolites of Fusarium moniliforme, a fungus that occurs widely in corn. Fumonisins causes leukoencephalomalacia in horses and pulmonary edema in swine and have been suggested as a possible cause of an increased incidence of neural tube defects among people living along the Texas-Mexico border. As part of an effort to determine levels of fumonisins in human food, a liquid chromatographic (LC) method was devised for determining fumonisin B1 (FB1) and the total hydrolysis product of FB1 (HB1) in tortillas. The method uses acetonitrile-0.1M phosphate buffer (pH 3; 1 + 1) extraction, solid-phase C18 cleanup, o-phthalaldehyde and 2-mercaptoethanol derivatization, and reversed-phase LC. Average recoveries from tortillas spiked with FB1 and HB1 at 250, 500, and 1000 ng/g were 86.5% for FB1 and 82.6% for HB1. Tortillas (54) and masa (8) from the Texas-Mexico border were analyzed for FB1 and HB1. Average amounts of FB1 and HB1 in tortillas were 187 and 82 ng/g, respectively. Average amounts of FB1 and HB1 in masas were 262 and 64 ng/g, respectively. The results show that fumonisin B1 and its hydrolysis product are present in tortillas consumed by a population experiencing an increased incidence of neural tube defects.
Carboxylic Acids↗