Biomedical subjects

S Nesheim

Publications and source records attributed to S Nesheim.

44 records · Page 3Linked to original sources

PubMedDate not supplied

Thin layer chromatographic method for determination of deoxynivalenol in wheat: collaborative study.

A collaborative study of a rapid method for the determination of deoxynivalenol (DON) in winter wheat was successfully completed. The method involves sample extraction with acetonitrile-water (84 + 16), cleanup using a disposable column of charcoal, Celite, and alumina, and detection by thin layer chromatography after spraying with an aluminum chloride solution. Each of the 15 collaborators analyzed 12 samples, 2 of which were naturally contaminated, and 10 to which DON was added, in duplicate, at levels of 0, 50, 100, 300, and 1000 ng/g. Average recoveries of DON ranged from 78 to 96% with repeatabilities of 30-64% and reproducibilities of 33-87%. The results of the study show that false positives were not a problem and that all of the analysts could detect DON at the 300 ng/g level or higher. The method has been adopted official first action.

Chromatography, Thin Layer↗

PubMedDate not supplied

Rapid quantitation and confirmation of aflatoxins in corn and peanut butter, using a disposable silica gel column, thin layer chromatography, and gas chromatography/mass spectrometry.

A simple, rapid, and solvent-efficient method for determining aflatoxins in corn and peanut butter is described. Aflatoxins B1, B2, G1, and G2 were extracted from 50 g sample with 200 mL methanol-water (85 + 15). A portion of the extract was diluted with 10% NaCl solution to a final concentration of 50% methanol, and then defatted with hexane. The aflatoxins were partitioned into chloroform. The chloroform solution was evaporated, and the residue was placed on a 0.5 g disposable silica gel column. The column was washed with 3 mL each of hexane, ethyl ether, and methylene chloride. Aflatoxins were eluted with 6 mL chloroform-acetone (9 + 1). The solvent was removed by evaporation on a steam bath, and the aflatoxins were determined using thin layer chromatography (TLC) with silica gel plates and a chloroform-acetone (9 + 1) developing solvent. Overall average recovery of aflatoxin B1 from corn was 82%, and the limit of determination was 2 ng/g. For mass spectrometric (MS) confirmation, aflatoxin B1 in the extract from 3 g sample (20 ng/g) was purified by TLC and applied by direct on-column injection at 40 degrees C into a 6 m fused silica capillary gas chromatographic column. The column was connected directly to the ion source. After injection, the temperature was rapidly raised to 250 degrees C, and the purified extract was analyzed by negative ion chemical ionization MS.

Aflatoxin B1↗

PubMedDate not supplied

Thin layer chromatographic determination of deoxynivalenol in wheat and corn.

A thin layer chromatographic (TLC) method for determining deoxynivalenol (DON) in corn and wheat was developed. DON is extracted from the grain with acetonitrile-water (84 + 16) and filtered through a column of mixed alumina-charcoal-Celite (0.5 g + 0.7 g + 0.3 g). The solvent is evaporated on a steam bath. Ethyl acetate is added to the residue and heated to dissolve DON. After cooling, the residue is transferred to a vial with additional ethyl acetate and is dissolved in CHCl3-acetonitrile (4 + 1) for TLC on an AlCl3-impregnated silica gel plate with CHCl3-acetone-isopropanol (8 + 1 + 1). The plate is heated in a 120 degrees C oven for 7 min; a blue fluorescent spot is produced under longwave ultraviolet light. DON is quantitated visually and/or fluorodensitometrically by comparison with reference standards. The minimum detectable amount of DON is ca 20 ng/spot. The limit of DON determination is ca 40 ng/g for wheat and 100 ng/g for corn. Recoveries of DON added to wheat and corn at 100, 500, and 1000 ng/g levels were 85, 93, and 88% and 77, 80, and 80%, respectively.

Chromatography, Thin Layer↗

PubMedDate not supplied

Deoxynivalenol in winter wheat: thin layer chromatographic method and survey.

A rapid method for the determination of deoxynivalenol (DON) in wheat was used to analyze 57 wheat samples collected from 4 midwestern states where the winter wheat crop was contaminated with Fusaria. The method involves sample extraction with acetonitrile-water (84 + 16), cleanup by charcoal-alumina column chromatography, and determination by thin layer chromatography (TLC), using an AlCl3 solution spray and heat to form a fluorescent derivative. Recoveries of DON added to wheat at levels as low as 0.2 micrograms/g averaged greater than 80%. DON was detected at an average level of 3.6 micrograms/g; the levels ranged from 0.2 to 9.0 micrograms/g in 54 of 57 of the wheat samples. The quantity of DON was, in general, proportional to the percentage of total damaged kernels (grade). The chemical identity of DON was confirmed by mass spectrometry after isolation with preparative TLC.

Chromatography, Thin Layer↗

PubMedDate not supplied

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↗

PubMedDate not supplied

Solvent-efficient thin-layer chromatographic method for the determination of aflatoxins B1, B2, G1, and G2 in corn and peanut products: collaborative study.

An interlaboratory study of a solvent-efficient thin-layer chromatographic (TLC) method for the determination of aflatoxins B1, B2, G1, and G2 was conducted in laboratories located in the United States, France, Tunisia, and Denmark. Eighteen artificially contaminated samples plus blanks of raw peanuts and peanut butter and corn containing varying amounts of aflatoxins B1, B2, G1, and G2 were distributed to participating laboratories. The method consists of elements of the U.S. Food and Drug Administration (FDA), Contaminants Branch (CB) (AOAC Method 968.22) and FDA, Best Foods (BF) (AOAC Method 970.45) methods with reduced requirements for solvents. Participating laboratories used either visual or densitometric techniques during the final determinative step. Statistical analysis of the data was performed to determine or confirm outliers and to compute repeatability and reproducibility of the method using either visual or densitometric techniques for the determinative step. Reported results from laboratories using a densitometer showed that, for corn, the relative standard deviation for repeatability (RSDr) for aflatoxin B1 ranged from 56.6 to 41.7% for contamination levels ranging from 5 to 50 ng/g. For raw peanuts and peanut butter, the RSDr values for aflatoxin B1 ranged from 21.3 to 37.3% and 65.9 to 42.1%, respectively, for the contamination levels ranging from 5 to 25 ng/g. RSDr ranges for aflatoxins B2, G1, and G2 were similar. For reproducibility (R), the RSDR ranges for aflatoxin B1 were 41.7-56.6%, 56.6-84.8%, and 26.4-37.3% for corn, peanut butter, and raw peanuts, respectively. Average recoveries for all aflatoxins at all levels were 95.3, 139.0, and 95.6% for corn, peanut butter, and raw peanuts, respectively. When analysts determined aflatoxin concentrations in corn by visual comparison to standards, the RSDr values for aflatoxin B1 were 47.8-11.4% for contamination levels ranging from 5 to 50 ng/g. For raw peanuts and peanut butter, the RSDr values for aflatoxin B1 were 76.3-12.6% and 33.4-8.8%, respectively, for the contamination levels ranging from 5 to 25 ng/g. RSDr values for aflatoxins B2, G1, and G2 were similar. The RSDR values for aflatoxin B1 were 34.6-90.2%, 45.5-59.3%, and 31.8-78.3% for corn, peanut butter, and raw peanuts, respectively. Average recoveries for all aflatoxins at all levels were 111.0, 157.6, and 92.3% for corn, peanut butter, and raw peanuts, respectively.(ABSTRACT TRUNCATED AT 400 WORDS)

Aflatoxin B1↗

PubMedDate not supplied

Reliability of mycotoxin assays--an update.

The precision parameters of the method-performance (collaborative) studies for mycotoxins published in the literature through 1991 have been recalculated on a uniform basis by following the International Union of Pure and Applied Chemistry protocol. About 80% of the 793 accepted assays for mycotoxins, almost all of which have been conducted by thin-layer chromatography (TLC), liquid chromatography (LC), and enzyme-linked immunosorbent assays (ELISA), exhibit relative standard deviations among laboratories (RSDR) that are less than 2 times the values predicted from the Horwitz equation: RSDR, % = 2(1-0.5log10C) where C is the concentration expressed as a decimal fraction. The precision of TLC and LC methods is about the same, but that of ELISA is somewhat poorer. For those commodities for which sufficient data exist to provide a meaningful comparison, the methods applied to cottonseed products have the best precision and corn the worst, with peanuts intermediate. Overall, however, the primary factor affecting RSDR is concentration, more or less independent of analyte, method, matrix, and age of the study. If it is assumed that the test results are normally distributed and that an RSDR of 50% is the point where effective control of the results begins to be lost (a value equivalent to the production of 2% false-negative values), then relying on the Horwitz curve, the limit of quantitative measurement is the single digit, i.e., 5, micrograms/kg (10(-9); ppb) concentration for solid food commodities. Such a value must be considered as a limit applicable to a single analyte, aflatoxin B1, and not as a mean, and not applicable to the sum of the individual components, each of whose associated standard deviation would lie in the unacceptable region. Enforcement of a 5 micrograms aflatoxin B1/kg limit, under the assumptions made, requires that a responsible manufacturer and a prudent regulator operate at opposite extremes of tolerance limits: e.g., the producer at 2 micrograms/kg and the consumer at 10. A proposed Codex "maximum level" of 0.05 micrograms aflatoxin M1/kg milk cannot be supported by the available data applied in an interlaboratory enforcement environment. These conclusions are also supported by an examination of the reported data from the ongoing, large-scale proficiency studies routinely performed by the American Oil Chemists' Society and the International Agency for Research on Cancer.

Databases, Factual↗

PubMedDate not supplied

Variability associated with analytical methods used to measure aflatoxin in agricultural commodities.

A total of 1019 analytical precision estimates obtained from method-performance (collaborative) studies for mycotoxins published through 1991 were sorted by type of variance measurement, type of analytical method, and type of agricultural commodity. Precision estimates for total aflatoxin were sorted into 2 precision measurements (among-laboratories and within-laboratory), 3 analytical methods (thin-layer chromatography [TLC], liquid chromatography [LC], and enzyme-linked immunosorbent assay [ELISA]), and 11 agricultural commodities. Sufficient data existed to study the analytical variability (precision) associated with 36 sorted combinations (of a possible 66). In all but one combination (within-laboratory, barley, and TLC), the variance (V) was a function of total aflatoxin concentration (C). A power function of the form V = aCb, where a and b are constants, describes the relationship between variance and aflatoxin concentration. The coefficients a and b were determined from regression analysis. When results were pooled across all agricultural commodities, LC had the lowest analytical variability while ELISA had the highest. For a given method, among-laboratories variability was approximately double the within-laboratory variability. These analytical variability estimates can be coupled with previously determined variability estimates of sampling and sample preparation to determine the performance associated with specific test procedures used to inspect agricultural commodities for aflatoxin.

Aflatoxins↗