Combination of total solids determined by oven drying and fat determined by Mojonnier extraction for measurement of solids-not-fat content of raw milk: collaborative study

Each of 9 laboratories analyzed 9 pairs of blind duplicate raw milk samples for fat, using the Mojonnier ether extraction method (16.E13-16.E17), and for total solids, using a new direct forced air oven method. Solids-not-fat was determined by subtracting percent fat from percent total solids. The solids-not-fat content of the samples tested in the collaborative study ranged from 8.48 to 9.36%. The average repeatability standard deviation (sr) and the average reproducibility standard deviation (SR) for the solids-not-fat method were 0.019 and 0.041, respectively. Average repeatability (RSDr) and reproducibility (RSDR) relative standard deviations were 0.218 and 0.466%, respectively. The mean repeatability value (r) was 0.055; the mean reproducibility value (R) was 0.117. The difference between milk total solids determined by direct forced air oven drying and milk fat determined by Mojonnier ether extraction has been approved for interim official first action for determination of solids-not-fat content of milk.     

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.     

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.     

Liquid chromatographic determination of penicillin V potassium in tablets: collaborative study

A liquid chromatographic method for the determination of penicillin V potassium in tablets was collaboratively studied by 7 laboratories. The method uses an octadecyl silane reverse-phase column, a mobile phase of acetonitrile-methanol-0.01 M potassium phosphate monobasic (21 + 4 + 75 v/v/v), photometric detection at 225 nm, and sulfadimethoxine as an internal standard. Each collaborator received 6 samples: powdered composites of 2 commercial tablet preparations and 1 synthetic tablet powder mixture, each with blind duplicates. The mean repeatability and reproducibility relative standard deviations for commercial samples were, respectively, 1.10 and 1.46% (250 mg dosage), and 0.84 and 2.82% (500 mg dosage). The average standard recovery from the synthetic formulation was 99.1%, with repeatability and reproducibility relative standard deviations of 1.30 and 3.66%, respectively. The method has been adopted official first action.     

Modified method for carbadox in feeds: collaborative study

The official first action method for carbadox in swine feed, 42.C01-42.C04, was modified in 2 respects. First, the samples were leached overnight at room temperature instead of boiled for 1 hr. This change avoided problems with overheating and excessive evaporation. Second, the dilution scheme for samples spiked with carbadox standard solution was changed to give absorbance values that were within the optimum working range of all types of spectrophotometers. The modified procedure was collaboratively studied by 21 laboratories. The repeatability standard deviation (sigma0) and reproducibility standard deviation (sigmax) were sigma0 = 0.00029% and sigmax = 0.00056% (8.9% of grand mean) for feeds containing 0.00617% carbadox; and sigma = 0.0012% and sigmax = 0.0019% (9.3% of grand mean) for feeds containing 0.0198% carbadox. The between-laboratory variance ratio was significant for feeds containing 0.0198% carbadox. The mean per cent of intent values for feeds containing 0.00617% carbadox and 0.0198% carbadox were 102% and 104%, respectively. In general, the statistical results were comparable to those previously obtained for the official first action method. Consequently, the modified procedure is not recommended as a replacement for the official first action method.     

Dry rehydratable film for enumeration of total aerobic bacteria in foods: collaborative study

A rehydratable dry-film plating procedure for aerobic plate counts has been compared to the standard agar plate method (966.23B and C, 15th ed.; 46.014-46.015, 14th ed.) in a collaborative study by 12 laboratories. Each laboratory analyzed the normal microflora of 3 samples in duplicate for 6 products. The aerobic plate counts ranged from 1.0 x 10(3) to 1.0 x 10(8) cfu/g. The products were flour, nuts, frozen raw shrimp, spice, frozen raw ground turkey, and frozen and refrigerated vegetables. Repeatability standard deviations of the 2 methods did not differ significantly for 13 of 18 test samples. For 1 shrimp and 2 turkey samples, the dry-film method had lower repeatability variances (P less than 0.05) and for 1 spice sample the agar method had lower repeatability variances (P less than 0.05). Relative standard deviations of repeatability were between 1.7 and 15.5% for the dry-film method and 1.2 and 16.0% for the agar method. Relative standard deviations of reproducibility ranged from 2.4 to 23.4% for the dry-film method and 2.3 to 18.8% for the agar method. The dry rehydratable film method has been adopted official first action for determination of the aerobic plate count.     

Sulfuric acid/hydrogen peroxide digestion and colorimetric a collaborative study

Seven laboratories participated in a collaborative study of a method for determination of phosphorus in meat and meat products. Samples are digested in sulfuric acid and hydrogen peroxide; digestion is complete in approximately 10 min. Phosphorus is determined by colorimetric analysis of a dilute aliquot of the sample digest. The collaborators analyzed 3 sets of blind duplicate samples from each of 6 classes of meat (U.S. Department of Agriculture classifications): smoked ham, water-added ham, canned ham, pork sausage, cooked sausage, and hamburger. The calibration curve was linear over the range of standard solutions prepared (phosphorus levels from 0.05 to 1.00%); levels in the collaborative study samples ranged from 0.10 to 0.30%. Standard deviations for repeatability (sr) and reproducibility (SR) ranged from 0.004 to 0.012 and 0.007 to 0.014, respectively. Corresponding relative standard deviations (RSDr and RSDR, respectively) ranged from 1.70 to 7.28% and 3.50 to 9.87%. Six laboratories analyzed samples by both the proposed method and AOAC method 24.016 (14th Ed.). One laboratory reported results by the proposed method only. Statistical evaluations indicated no significant difference between the 2 methods. The method has been adopted official first action by AOAC.     

Liquid chromatographic determination of methyldopa and methyldopa-thiazide combinations in tablets: collaborative study

A reverse phase liquid chromatographic method for the determination of methyldopa, methyldopa-hydrochlorothiazide, and methyldopachlorothiazide in tablets was collaboratively studied by 8 laboratories. Each collaborator received 20 samples that included drug substance, synthetic and commercial tablet compositions. The overall repeatability and reproducibility standard deviations for commercial tablets were 1.11 and 1.75% for methyldopa, 0.96 and 1.62% for chlorothiazide, and 1.21 and 2.15% for hydrochlorothiazide, respectively. The overall recoveries of methyldopa, chlorothiazide, and hydrochlorothiazide added to synthetic tablets were 100.78, 100.70, and 101.34%, respectively. The method has been adopted official first action.     

Determination of fat in raw milk: comparison of mojonnier (ether extraction) and Gerber method

Results are compared for the approved Gerber method that specifies a TC 11.07 mL pipet and 2 modifications that specify a TC 10.77 mL pipet or a weighed 11.125 g sample of milk rather than a pipetted sample. All results were also compared with results obtained with the Mojonnier method for determining the fat content of raw milk. The standard deviation is somewhat lower for the Mojonnier method than for any of the Gerber methods; among the Gerber methods, the standard deviations were lower for the TC 10.77 mL and 11.125 g modifications. It is recommended that one of these modifications replace the current specified TC 11.07 mL pipet method.     

Enumeration of total bacteria and coliforms in milk by dry rehydratable film methods: collaborative study

Eleven laboratories participated in a collaborative study to compare the dry rehydratable film (Petrifilm SM and Petrifilm VRB) methods, respectively, to the standard plate count (SPC) and violet red bile agar (VRBA) standard methods for estimation of total bacteria and coliform counts in raw and homogenized pasteurized milk. Each laboratory analyzed 16 samples (8 different samples in blind duplicate) for total count by both the SPC and Petrifilm SM methods. A second set of 16 samples was analyzed by the VRBA and Petrifilm VRB methods. The repeatability standard deviations (the square root of the between-replicates variance) of the SPC, Petrifilm SM, VRBA, and Petrifilm VRB methods were 0.05104, 0.0444, 0.14606, and 0.13806, respectively; the reproducibility standard deviations were 0.7197, 0.06380, 0.15326, and 0.13806, respectively. The difference between the mean log10 SPC and the mean log10 Petrifilm SM results was 0.027. For the VRBA and Petrifilm VRB methods, the mean log10 difference was 0.013. These results generally indicate the suitability of the dry rehydratable film methods as alternatives to the SPC and VRBA methods for milk samples. The methods have been adopted official first action.     

Liquid chromatographic method for multiresidue determination of benzimidazoles in beef liver and muscle: collaborative study

A liquid chromatographic method for determination of thiabendazole, 5-hydroxythiabendazole, oxfendazole, mebendazole (MBZ), and fenbendazole (FBZ) in cattle liver and muscle was collaboratively studied in 7 laboratories in 1986. For blind fortified samples containing 800 ppb FBZ, average recovery and relative standard deviations for repeatability and reproducibility (RSDr and RSDR) based on results from 6 of the participating laboratories were 83%, 12.7%, and 14.0%, respectively. Recoveries of FBZ from incurred liver samples were more variable. Recoveries of MBZ from livers fortified at the 100 ppb level were encouraging; however, the drug levels were too low in the incurred samples used for MBZ studies. Except for FBZ and MBZ in liver, the study data were not satisfactory. The method has been adopted official first action by AOAC for determination of 800-1600 ppb fenbendazole in liver. The analysis should be repeated using a smaller sample size when initial analyses show levels greater than 1600 ppb FBZ.     

Gas chromatographic-chemiluminescence method for determination of volatile N-nitrosamines in minced fish-meat and surimi-meat frankfurters: collaborative study

A collaborative study was carried out on a solid-phase extraction method for separating volatile N-nitrosamines, particularly N-nitrosodimethylamine (NDMA), from combination minced fish or surimi-meat frankfurters with detection by gas chromatography-chemiluminescence (thermal energy analyzer). The results from the 10 collaborators were evaluated using the most recent AOAC guidelines for determining outliers and for the analysis of variance. For NDMA, repeatability standard deviations, sr, ranged from 0.56 to 2.25; repeatability relative standard deviations, RSDr, ranged from 8.9 to 11.5%. Reproducibility standard deviations, SR, for NDMA ranged from 1.40 to 6.49, and reproducibility relative standard deviations, RSDR, ranged from 24.2 to 28.9%. Our data compared favorably to the reproducibility (RSDR) curve of Horwitz. The method has been adopted official first action by AOAC.     

Flame atomic absorption spectrometric determination of serum zinc: collaborative study

A method for determining serum zinc concentrations was collaboratively studied by 9 laboratories. Serum samples in polypropylene tubes are directly diluted with 0.03% Brij 35, and zinc is determined by flame atomic absorption spectrophotometry (AAS). A certified standard reference material serves as an external control. Repeatability relative standard deviations (RSDr) were 1.2 and 11.4% and reproducibility relative standard deviations (RSDR) were 6.1 and 12.9% for serum zinc concentrations of 6.36 and 0.63 micrograms/mL, respectively. The method has been approved interim official first action by AOAC.     

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.     

Gas chromatographic-thermal energy analysis method for N-nitrosodibutylamine in latex infant pacifiers: collaborative study

Each of 5 collaborating laboratories determined volatile N-nitrosamines in 3 blind quadruplicate sets of latex rubber infant pacifier samples, using a gas chromatographic-thermal energy analysis (GC-TEA) method. Volatile N-nitrosamines are extracted from cut-up pacifier nipples with CH2Cl2. The extract is concentrated and subjected to high temperature purge and trap, and the nitrosamines are eluted from the trap and determined by GC-TEA. N-Nitrosodibutylamine (NDBA) was the only nitrosamine found in sufficient concentration to allow analysis. NDBA concentrations of the 3 sets of samples were 82.6, 21.0, and 7.12 ng/g rubber. The repeatability relative standard deviations ranged from 7.46 to 24.0% and the reproducibility relative standard deviations from 7.46 to 29.2%. The minimum detectable level of NDBA by this method is 3.6 ng/g rubber. The method has been adopted official first action.     

Determination of nitrogen content in milk by the Kjeldahl method using copper sulfate: interlaboratory study

Copper sulfate was substituted for mercury as the catalyst in the International Dairy Federation (IDF) Standard 20A:1986 method for the determination of nitrogen content in milk. The substitution was supported by results obtained in an interlaboratory study by 24 laboratories in 12 countries. Each laboratory analyzed 12 test samples of milk as blind duplicates in a double split level design with high, medium, and low nitrogen concentrations. The method protocol requires the concurrent analyses of an ammonium salt solution and a tryptophan solution as internal quality control standards with a minimum nitrogen recovery between 99 and 100% for the former and at least 98% for the latter. The repeatability and reproducibility relative standard deviations are 0.5 and 1%, respectively, for the range 0.35-0.70 g N/100 g. The performance of the laboratories that did not meet the required quality control specifications was clearly poorer than that of those that did meet the specifications.     

Use of electrochemical biosensor and gas chromatography for determination of dichlorvos in wheat

Two methods for the determination of dichlorvos in durum wheat by electrochemical assay and gas chromatography, respectively, have been developed. Dichlorvos, an organophosphorus anticholinesterase pesticide, was extracted from wheat with hexane, and the filtered extract was directly analyzed by gas chromatography with nitrogen-phosphorus flame detection (NPD). Recoveries of dichlorvos from milled wheat spiked at 0.25-1.5 microg/g ranged from 96.5 to 100.9%, and the limit of detection was 0.02 microg/g. The electrochemical assay was based on the detection of choline, the acetylcholinesterase product, via a choline oxidase biosensor. An aliquot of the filtered hexane extract was partitioned with phosphate buffer solution, and the organic layer was evaporated prior to electrochemical analysis. A limit of detection of 0.05 microg/g of dichlorvos was obtained with mean recoveries of 97-103% at spiking levels of 0.25-1.5 microg/g. A good correlation (r = 0.9919) was found between the results obtained with the electrochemical and those obtained with the gas chromatographic methods. The electrochemical method was peer-validated in two laboratories that analyzed 10 blind samples (5 duplicates), including a blank and 4 spiked samples with dichlorvos at levels of 0.25, 0.60, 1.00, and 1.50 microg/g. Within-laboratory repeatability (RSDr) and between-laboratory reproducibility (RSDR) ranged from 5.5 to 7.8% and from 9.9 to 17.6%, respectively.     

In vitro assay for protein digestibility: interlaboratory study

True protein digestibilities of 17 protein sources were estimated by 6 laboratories using an in vitro, 3-enzyme digestion system in a pH stat. Samples from animal, vegetable, and mixed food sources were freeze-dried (if not already dried), ground, mixed, and shipped to each collaborator along with a sodium caseinate standard and trypsin, chymotrypsin, and peptidase. The uptake of titrant during enzymatic digestion was used to calculate estimates of digestibility. Digestibilities ranged from 100% for casein to 89.9% for whole wheat cereal. Mean relative standard deviations for repeatability were 1.4% for rolled oats and less than 1% for the remaining 16 samples. Mean relative standard deviations for reproducibility ranged from 5.0 to 0.8%; values were less than 2.5% for 13 of the 17 samples.     

Automated method for the determination of niacin and niacinamide in cereal products: collaborative study.

In a collaborative study, an automated method for the determination of niacin and niacinamide in cereal products was compared with the official final action microbiological (43.121-43.125) and chemical (43.044-43.046) methods. Ten samples of cereal products, including enriched flour, yeast-leavened baked products, fortified breakfast cereals, and baked pet food products, were submitted to 14 laboratories. Nine laboratories reported values by the automated method, 6 reported values by the microbiological method, and 7 reported values by the chemical method. The results from the microbiological method were not subjected to analysis of variance because of the unusually large between-laboratory variation. The between-laboratory coefficients of variation for the automated and chemical methods were 10.90 and 10.18%, on the basis of results from 7 and 4 laboratories, respectively. There was no significant (p greater than 0.05) difference between methods when results from the 4 laboratories who used both methods were compared. The automated chemical method has been adopted as official first action.     

Liquid chromatographic determination of colchicine in pharmaceuticals: collaborative study

A liquid chromatographic (LC) method for the determination of colchicine in pharmaceutical dosage forms and the bulk drug was evaluated in an interlaboratory study which included 13 participating laboratories. The method involves extraction (or dissolution) of the active ingredient with methanol-water (1 + 1), followed by filtration of the extract and reverse phase LC using an octylsilane bonded phase column and UV detection at 254 nm. The mobile phase consists of a methanol-phosphate buffer mixture (pH = 5.5). Three commercial tablet formulations (0.5-0.6 mg colchicine/tablet), 1 synthetic injectable preparation (0.510 mg colchicine/mL), and 1 bulk drug sample were assayed in duplicate by the proposed method. The reproducibility and repeatability standard deviations based on nonpooled results for each sample ranged from 0.0062 mg/mL to 0.0147 mg/tablet and from 0.0037 mg/mL to 0.0127 mg/tablet, respectively; the corresponding coefficients of variation ranged from 1.21 to 2.54% and from 0.73 to 2.19%, respectively. The mean recovery from the synthetic injectable formulation was 100.0%. The method has been adopted official first action.