High pressure liquid chromatographic determination of carbohydrates in food products: evaluation of method

A high pressure liquid chromatographic (HPLC) method for determining 5 common carbohydrates in food products was evaluated. Reproducibility data were generated showing a relative standard deviation of 2.8%. Recovery studies on a variety of foods gave an average recovery of 98.8%. The HPLC data for 3 varieties of ready-to-eat cereals were compared with data from 4 independent laboratories using current AOAC chemical methods. The HPLC mean values differed from the chemical mean values by 3.2%.     

International mycotoxin check sample program: Part I. Report on laboratory performance for determination of aflatoxins B1, B2, G1, and G2 in raw peanut meal, deoiled peanut meal, and yellow corn meal

Three aflatoxin-contaminated samples (raw peanut meal, deoiled peanut meal, and yellow corn meal) were analyzed by 121 laboratories in 31 countries. Sufficient data were obtained to permit a statistical comparison of the performance of laboratories using the BF, CB, and EEC methods and those using high performance liquid chromatography (HPLC) for quantitation. No significant differences were found between means for laboratories using these 4 methods for the analysis of raw peanut meal or yellow corn meal. However, for deoiled peanut meal, means were significantly different for laboratories using the BF method compared with the CB or EEC methods for B1 and B2, and for laboratories using the CB method compared with HPLC methods for G2.     

Analysis of fat-soluble vitamins. XXII. High performance liquid chromatographic determination of vitamin D in concentrates. Collaborative study

A collaborative study was carried out which compared the official chemical method (43.B14-43.B24), the HPLC method according to Hofsass et al. including maleic anhydride treatment, and the HLPC procedure according to De Vries et al. for vitamin D concentrates. A total of 396 samples were distributed to 33 collaborators for analysis. Five laboratories performed both the chemical and the HPLC methods. Five laboratories performed the Hofsass method and 16 laboratories performed the De Vries method. The results for the chemical method agreed with the theoretical values for the samples, and the standard deviation was comparable to that obtained in previous AOAC collaborative studies. Collaborative results for the Hofsass method were low. In addition, incorrect use of a fixed conversion factor (1/0.586) and necessity of a double chromatogrpahic system on a non-treated and a treated vitamin D sample reduce the effectiveness of the method. There were no adverse reactions to the DE Vries HPLC method. It is recommended that the method be adopted official first action as an alternative procedure for determining vitamin D in concentrates, excluding powders containing irradiated 7-dehydrocholesterol.     

Automated methods for determination of fat and moisture in meat and poultry products: collaborative study

A collaborative study was conducted to compare automated methods for rapid determination of fat and moisture in meat and poultry products with the official AOAC solvent extraction and forced-air oven methods, respectively. Fourteen products were tested, with fat and moisture contents ranging from 2 to 43% and 44 to 74%, respectively. Eight of the collaborating laboratories analyzed the products by using a moisture/fat analyzer; 4 laboratories used the AOAC methods. Standard deviations for within-laboratory repeatability, between-laboratory reproducibility, and bias for each product indicated that the rapid methods were acceptable. The moisture/fat analyzer methods have been adopted official first action for fat and moisture analyses in meat and poultry products.     

Evaluation of laboratory performance of the AOAC method for PSP toxin in shellfish

Laboratory performance of the official AOAC method for paralytic shellfish poison (PSP) toxin in shellfish was evaluated. Two series of naturally toxic shellfish split samples were distributed (15 in 1979 and 19 in 1982) to state shellfish-monitoring laboratories which participate in the National Shellfish Sanitation Program. The laboratories performed bioassays on duplicate 100 g portions of each 220 g split sample. Bioassays were consistent among the laboratories and compared favorably with those of previous studies.     

Comparison of two methods for determination of total solids content of milk: collaborative study

Ten laboratories analyzed 9 pairs of blind duplicate raw milk samples for total solids. A direct forced air oven method (4 h at 100 degrees C) and a modification of the AOAC predry method (16.032) were used. Preliminary evaluation of the modified AOAC method indicated that blank determinations were necessary. Total solids content ranged from 12.0 to 14.6%. Average repeatability standard deviations (Sr) of the direct forced air oven and modified AOAC methods were 0.019 and 0.017, respectively. Average reproducibility standard deviations (SR) of the direct forced air oven and the modified AOAC methods were 0.042 and 0.047, respectively. Average repeatability relative standard deviations (RSDr) for the direct forced air oven and the modified AOAC methods were 0.149 and 0.136%, respectively; average reproducibility relative standard deviations (RSDR) were 0.327 and 0.370%, respectively. Mean repeatability values (r) and reproducibility values (R) were 0.054 and 0.118 for the direct forced air oven method and 0.049 and 0.133 for the modified AOAC method, respectively. The mean test result of the direct forced air oven method (12.7293%) was comparable to that for the modified AOAC method (12.7273%). The modification of AOAC method 16.032 and the direct forced air oven method have been approved interim official first action.     

Immunodiffusion screening method for detection of motile Salmonella in foods: collaborative study

A collaborative study was performed to validate the performance of the 1-2 TEST for detection of motile salmonellae in foods. Detection is based on observation of an immobilized band of cells. Twenty-three laboratories participated in the study. The 1-2 TEST (immunodiffusion test) was compared with the standard culture procedure (BAM/AOAC; FDA Bacteriological Analytical Manual) for detection of Salmonella in 6 food types: ground black pepper, soy flour, dried whole egg, milk chocolate, nonfat dry milk, and raw deboned turkey. Uninoculated and inoculated samples were included in each food group analyzed. After the tests on the 6 foods were completed, analysis of the data for turkey and soy flour showed that certain collaborators obtained data inconsistent with the data from the majority of collaborators. No specific method deviations to account for the inconsistencies were reported by those collaborators, so the collaborative testing of these 2 foods was repeated. Analysis of data for pepper, chocolate, nonfat dry milk, dried whole egg, and the second set of soy flour and turkey indicated 96.1% agreement between the BAM/AOAC and immunodiffusion test methods. The false negative rates for the immunodiffusion test and BAM/AOAC methods were 3.6 and 1.7%, respectively. There was no significant difference in the productivity of the immunodiffusion test and BAM/AOAC method at the 5% level for any of the 6 foods. The immunodiffusion screening method has been approved official first action for detection of motile Salmonella in foods.     

Collaborative study of the gravimetric method for the determination of mercury in chlorinated pesticide formulations.

The official first action AOAC gravimetric method for mercury, 6.C01-6.C03, and the official CIPAC titrimetric method, M2.3, were collaboratively studied by 4 CIPAC laboratories, using the same 4 samples analyzed by AOAC collaboration in 1972. Averages at the 3.7 and 3.4% level were the same by both methods whereas the CIPAC method produced, on the average, 4% higher results, all levels included. The AOAC method performs well in the presence of copper and chlorinated pesticides and is less complex to use than the present CIPAC method for mercury in these mixtures. The method has been adopted as official final action.     

Evaluation of methods used to determine ochratoxin A in coffee beans

A comparative study was conducted to evaluate four previously reported methods that proved to have a recovery greater than 80% for the determination of different levels of ochratoxin A (OTA) in green and roasted coffee beans and to select an accurate, sensitive, and less-expensive technique between the existing methods. The results indicated that the Association of Official Analytical Chemists (AOAC) official method for the extraction of OTA in green coffee and determination by high-performance liquid chromatography (HPLC) is recommended as an efficient method for the routine analyses of OTA in green and ground roasted coffee beans. This method proved to be an accurate, sensitive, and less-expensive method that employs routine materials and available equipment. Although the immunoaffinity column/HPLC procedure tested showed a significantly higher percentage than the AOAC recommended method, it is recommended for use in processed coffee beans where low concentrations of OTA may be expected to be detected.     

Hydrophobic grid membrane filter/MUG method for total coliform and Escherichia coli enumeration in foods: collaborative study

Twenty-four laboratories participated in a collaborative study to validate a hydrophobic grid membrane filter (HGMF) method incorporating the use of 4-methylumbelliferyl-beta-D-glucuronide (MUG) for enumeration of total coliform and Escherichia coli bacteria in foods by comparing its performance against the AOAC 3-tube MPN method (46.013-46.016). Raw milk, raw ground poultry, whole egg powder, cheese powder, and ground black pepper were included in the study. The total coliform methods did not differ significantly, except that the 3-tube method detected a significantly higher level of total coliforms than did the HGMF method in the ground black pepper. Conversely, the HGMF/MUG E. coli method detected significantly higher numbers of E. coli present in the egg powder, cheese powder, and ground black pepper samples, while not differing significantly from the 3-tube method for the raw milk and raw ground poultry samples. The overall confirmation rate of MUG-positive colonies isolated using the HGMF method was 99.5%. The hydrophobic grid membrane filter/MUG method has been adopted official first action as an additional method to AOAC official final action method 46.030-46.034.     

Improved hydrophobic grid membrane filter method, using EF-18 agar, for detection of Salmonella in foods: collaborative study

A collaborative study was carried out in 30 laboratories to validate improvements to the official final action hydrophobic grid membrane filter (HGMF) screening method for Salmonella in foods, 985.42, by comparing the performance of the improved HGMF method against that of the AOAC/BAM conventional culture method. Six products were included in the collaborative study: milk chocolate, raw deboned poultry meat, black pepper, soy flour, egg yolk powder, and nonfat dry milk. The raw deboned poultry meat was naturally contaminated with Salmonella, and the remaining 5 products were each inoculated in advance with low levels of individual Salmonella serotypes. The AOAC/BAM method produced 11 false negative results and the improved HGMF method produced 18 false negative results. The improved HGMF Salmonella method has been approved interim official first action for all foods to replace the HGMF official final action method, 985.42.     

Fluorometric determination of histamine in tuna: collaborative study

Six samples of canned tuna, albacore, yellowfin, and skipjack, in water or oil pack were analyzed in duplicate by a fluorometric method and the AOAC colorimetric method. For the fluorometric method, recoveries of histamine added to acceptable tuna averaged 99% with a range of 91 to 107%. Agreement between laboratories for the analyses of decomposed tuna containing 20-200 mg histamine/100 g sample was excellent. Results from the fluorometric method are comparable with those from the AOAC colorimetric method; the fluorometric method has been adopted as official first action.     

Hydrophobic grid membrane filter method for aerobic plate count in foods: collaborative study

Twenty-one laboratories participated in a collaborative study to validate a hydrophobic grid membrane filter (HGMF) method for aerobic plate count by comparing its performance against the AOAC/APHA pour plate method. Raw milk, raw poultry, whole egg powder, flours, and spices were included in the study. Counts obtained by the HGMF and pour plate methods did not differ significantly, except in the case of whole egg powder, for which the HGMF method produced significantly higher counts. The hydrophobic grid membrane filter method for aerobic plate count in foods has been adopted official first action.     

Dry rehydratable film for enumeration of total coliforms and Escherichia coli in foods: collaborative study

Rehydratable dry-film plating methods for total coliforms and Escherichia coli in foods have been compared to the AOAC most probable number methods. Fourteen laboratories participated in the collaborative study. Three coliform and E. coli levels in 6 samples of 4 product types (flour, nuts, cheese, and beef with gravy) and in 3 samples of 2 product types (mushrooms and raw turkey) were tested in duplicate by the participants. The mean log counts for the 3 methods were comparable. In general, the repeatability and reproducibility variances of the plating methods were as good as or better than that of the MPN method. The method has been adopted official first action by AOAC.     

Comparison of high pressure liquid chromatographic and chemical methods for vitamin D3 concentrates. II. Collaborative study.

A collaborative study was carried out which compared the official chemical method, 43.B14-43.B24, the official rat bioassay, 43.165, and the high pressure liquid chromatographic method for vitamin D3 resin, vitamin D3 resin in oil, and dry concentrate. A total of 340 samples were distributed to 17 collaborators for analysis. Five laboratories performed both the chemical and HPLC methods on 5 sets of blind duplicates. A 2-way analysis of variance comparing both methods for each sample showed a significant (P less than 0.01) difference between methods only for Sample 5. When the 2 methods were compared over all the samples, no significant (P less than 0.05) difference was found. Except for Sample 5, there were no differences in the repeatability of the methods. Per cent recoveries on Sample 3, which contained exactly 0.200 X 10(6) IU/g, showed 98.2% for the chemical method and 100.6% for the HPLC method for the 5 laboratories that performed both methods. The assay results of the HPLC and chemical methods are in good agreement with those found by the biological assay on Samples 1-4, but not for Sample 5. Evidence indicates that Sample 5 degraded partially to isotachysterol, and while the HPLC method yielded a reasonable value on this material, the chemical method erroneously showed full potency. An amendment is included for the collaboratively studied HPLC method which detects and eliminates 5,6-trans-vitamin D3, a possible interferant.     

Enzymatic-ultraviolet method for measuring lactose in milk: collaborative study

Collaborators in 8 dairy and food industry laboratories performed one lactose determination on each of 8 unknown samples of milk, lowfat milk, or skim milk, as 3 pairs of blind duplicates. Two known samples were provided to gain experience prior to analysis of the unknown samples. All of the above samples were also analyzed for lactose content by the official AOAC gravimetric method (16.507) by a commercial laboratory. From the overall mean of results on all samples, determinations by the enzymatic method averaged 0.49% lower than by the AOAC method. This difference was significant by the t-test (P = 0.05), which indicated a lack of agreement between the compared methods in determining lactose content. Standard deviations were similar for the 3 sets of blind duplicates which ranged between 3.67 and 4.55% lactose content. F-values revealed that variations between means obtained by laboratories differed significantly as compared with variations within laboratory means. The method has been adopted official first action.     

Direct aqueous injection-liquid chromatography with post-column derivatization for determination of N-methylcarbamoyloximes and N-methylcarbamates in finished drinking water: collaborative study

An interlaboratory method validation study was conducted on EPA Method 531.1, Measurement of N-Methylcarbamoyloximes and N-Methylcarbamates in Water by Direct Aqueous Injection HPLC with Post Column Derivatization, to determine the precision and mean recovery for determination of 10 carbamate pesticide compounds in reagent water and in finished drinking waters. The study design was based on Youden's nonreplicate plan for collaborative tests of analytical methods. The waters were spiked with 10 carbamate pesticides at 6 concentration levels, as 3 Youden pairs. Eight laboratories analyzed the samples by direct aqueous injection, with separation by reverse-phase liquid chromatography and post-column hydrolysis of the carbamates and carbamoyloximes to methylamine, followed by reaction of the methylamine with o-phthalaldehyde and 2-mercaptoethanol using fluorescence detection. Results were analyzed using an EPA computer program, which measured precision and recovery for each of the 10 compounds and compared the performance of the method between water types. The method was acceptable for all analytes tested. After removal of a nonrepresentative data set for aldicarb sulfoxide, no matrix effects were observed; the statistics for the pooled drinking waters were not significantly different from the statistics for the reagent waters. The method has been adopted official first action by AOAC.     

High-pressure liquid chromatographic determination of vitamin D3 in resins, oils, dry concentrates, and dry concentrates containing vitamin A.

A high-pressur liquid chromatographic (HPLC) procedure has been developed for the determination of vitamin D3 in resins, oils, dry concentrates, and dry concentrates containing vitamin A. Specificity of the method for vitamin D3 in the presence of isomers and other related constituents was shown by ultimate recovery of the vitamin D3 and the individual constituents and their characterization by other methods such as ultraviolet spectrophotometry. Precision and accuracy are within 2%, and as many as 20 determinations may be completed in a working day. Excellent agreement with the official AOAC biological assay was found. A comparison of the response of isomers of vitamin D3 by the HPLC method vs. other instrumental and chemical procedures shows HPLC to be the most specific method for determining the bioactive isomers.     

Comparison of HgO and CuSO4 as digestion catalysts in manual Kjeldahl determination of crude protein in animal feeds: collaborative study

The official AOAC manual Kjeldahl method for determining crude protein in animal feeds, 7.015, uses HgO as a catalyst in the digestion step. Because of environmental considerations, there is considerable interest in alternative catalysts. A collaborative study compares the official HgO-catalyzed method and an alternative using CuSO4. Fifty-four samples consisting of blind duplicates of closely matched pairs, representing a range of animal feed materials and 2 standard materials, were analyzed once by each method. Results were returned by 22 laboratories. Means and standard deviations between methods were comparable. The CuSO4-catalyzed method has been adopted official first action.     

Leak in evaporated milk by anodic stripping voltammetry and atomic absorption spectrophotometry: cooperative interlaboratory study

The results of a cooperative study on the determination of lead in evaporated milk, using a double blind referee technique, are reported. This study was designed to determine the normal variability of methods currently used for lead analysis by canned food industry laboratories. Twenty-three laboratories participated in this study. Each laboratory was instructed to use atomic absorption spectrophotometry (AOAC 25.065), anodic stripping voltammetry, or carbon rod atomic absorption spectrophotometry. Overall, the results appear to be in close agreement with the spiking levels. The coefficient of variation for all laboratories was 36.0% at the 0.15 ppm lead level and 16.8% at the 0.40 ppm lead level.