Water-soluble vitamins

Simultaneous Determination of Vitamins.--Klejdus et al. described a simultaneous determination of 10 water- and 10 fat-soluble vitamins in pharmaceutical preparations by liquid chromatography-diode-array detection (LC-DAD). A combined isocratic and linear gradient allowed separation of vitamins in 3 distinct groups: polar, low-polar, and nonpolar. The method was applied to pharmaceutical preparations, fortified powdered drinks, and food samples, for which results were in good agreement with values claimed. Heudi et al. described a separation of 9 water-soluble vitamins by LC-UV. The method was applied for the quantification of vitamins in polyvitaminated premixes used for the fortification of infant nutrition products. The repeatability of the method was evaluated at different concentration levels and coefficients of variation were <6.5%. The concentrations of vitamins found in premixes with the method were comparable to the values declared. A disadvantage of the methods mentioned above is that sample composition has to be known in advance. According to European legislation, for example, foods might be fortified with riboflavin phosphate or thiamin phosphate, vitamers which are not included in the simultaneous separations described. Vitamin B2.--Vi?as et al. elaborated an LC analysis of riboflavin vitamers in foods. Vitamin B2 can be found in nature as the free riboflavin, but in most biological materials it occurs predominantly in the form of 2 coenzymes, flavin mononucleotide (FMN) and flavin-adenine dinucleotide (FAD). Several methods usually involve the conversion of these coenzymes into free riboflavin before quantification of total riboflavin. According to the authors, there is growing interest to know flavin composition of foods. The described method separates the individual vitamers isocratically. Accuracy of the method is tested with 2 certified reference materials (CRMs). Vitamin B5.-Methods for the determination of vitamin B5 in foods are limited because of their low sensitivity and poor selectivity. Pakin et al. proposed a post-column derivatization of pantothenic acid as a fluorescent compound and used this principle in a specific and sensitive method for the determination of free and bound pantothenic acid in a large variety of foods. A French laboratory invited European laboratories to participate in a series of collaborative studies for this method, which will be carried out in 2005/2006. A more sophisticated method was described by Mittermayer et al. They developed an LC-mass spectrometry (LC/MS) method for the determination of vitamin B5 in a wide range of fortified food products. Application of the method to various samples showed consistent results with those obtained by microbiology. Vitamin B6.-Method 2004.07, an LC method for the analysis of vitamin B6 in reconstituted infant formula, was published by Mann et al. In contrast with this method, which quantifies vitamin B6 after converting the phosphorylated and free vitamers into pyridoxine, Vi?as et al. published an LC method which determines 6 vitamin B6 related compounds, the 3 B6 vitamers, their corresponding phosphorylated esters, and a metabolite. Accuracy was determined using 2 CRMs. Results were within the certified ranges. Vitamin C.-Franke et al. described an extensive study to vitamin C and flavonoid levels of fruits and vegetables consumed in Hawaii. Vitamin C was determined by measuring ascorbic acid in its reduced state by LC and coulometric detection along with UV absorbance detection at 245 nm. No attempts were made to assess levels of dehydroascorbic acid. Most recent research revealed that cell uptake of dehydroascorbic acid is unlikely to play a major role, which may explain the very low vitamin C activity of orally administered L-dehydroascorbic acid in rats. The food levels found by Franke et al. are variably lower, higher, or equal in comparison to other studies. Iwase described a method for the determination of ascorbic acid in foods using L-methionine for the pre-analysis sample stabilization. Electrochemical detection was used for the quantification. Traditionally, metaphosphoric acid was proven to be a useful dissolving agent for the determination of ascorbic acid. However, it dissolves in water very slowly, it is hygroscopic, and accurate weighing is not easy. Adjustment at pH 2-3 takes a long time. It appeared to be possible to replace metaphosphoric acid by 0.2% phosphoric acid. Methionine played an important role on the stability of ascorbic acid. The method seemed to be applicable to the routine analysis of ascorbic acid in foods. Folic Acid.-Microbiological analysis of total folate in foods is often considered as the golden standard compared to other methods based on, for example, LC. Koontz et al. showed results of total folate concentrations measured by microbiological assay in a variety of foods. Samples were submitted in a routine manner to experienced laboratories that regularly perform folate analysis fee-for-service basis in the United States. Each laboratory reported the use of a microbiological method similar to the AOAC Official Method for the determination of folic acid. Striking was, the use of 3 different pH extraction conditions by 4 laboratories. Only one laboratory reported using a tri-enzyme extraction. Results were evaluated. Results for folic acid fortified foods had considerably lower between-laboratory variation, 9-11%, versus >45% for other foods. Mean total folate ranged from 14 to 279 microg/100 g for a mixed vegetable reference material, from 5 to 70 microg/100 g for strawberries, and from 28 to 81 microg/100 g for wholemeal flour. One should realize a large variation in results, which might be caused by slight modifications in the microbiological analysis of total folate in foods or the analysis in various (unfortified) food matrixes. Furthermore, optimal combination of enzymes and reaction conditions may vary depending on the composition of the food. Padrangi and Laborde showed recently that treatment with alpha-amylase had no significant effect on measured folate in spinach, although addition of protease significantly increased the release of folate. LC/MS applications gain increasing attention because of their specificity. Rychlik used stable isotope dilution assays for the determination of the folate content of broccoli and bread. Compared to data in the literature and food data bases, amounts were significantly lower. Pawlosky et al., however, found comparable values for 5-methyltetrahydrofolic acid and folic acid by HPLC analysis with fluorescent detection and HPLC/MS. Among samples analyzed were CRMs and broccoli. Besides folic acid, other water-soluble vitamins were also determined by LC/MS/MS by Leporati et al. The method was applied to the quantitative analysis of the natural content of vitamins in typical Italian pasta samples, as well as in fortified pasta samples produced for the U.S. market. Biotin.-A paper from Staggs et al. included the assertion that existing biotin data in food composition tables are inaccurate because the majority are based on bioassays with all relevant disadvantages. Data in most cases are overestimated with consequences for recommendations for dietary biotin intake. An HPLC/avidin-binding assay was used to analyze 87 foods to support the hypothesis mentioned.     

Microbiological assay-trienzyme procedure for total folates in cereals and cereal foods: collaborative study

In 1996, U.S. Food and Drug Administration regulations mandated the fortification of enriched cereal-grain products with folic acid, thereby emphasizing the need for validated methods for total folates in foods, particularly cereal products. The AOAC Official Methods (944.12, 960.46) currently used for the analysis of folate in foods for compliance purposes are microbiological methods. When the fortification regulations were finalized, no Official AOAC or Approved AACC methods for folate in cereal-grain products were in place. The AOAC Official Method (992.05) for folic acid in infant formula does not incorporate important improvements in the extraction procedure and was not considered suitable for the analysis of folates in foods in general. A microbiological assay protocol using a trienzyme extraction procedure was prepared and submitted for comments to 40 laboratories with recognized experience in folate analysis. On the basis of comments, the method was revised to have the conjugase (gamma-glutamyl-carboxy-peptidase) treatment follow a protease treatment, to include the use of cryoprotected inoculum, and to include the spectroscopic standardization of the standard and optional use of microtiter plates. Thirteen laboratories participated in a collaborative study of 10 required and 10 optional cereal-grain products, including flour, bread, cookies, baking mixes, and ready-to-eat breakfast cereals. The majority of the participating laboratories performed the assay by the standard test tube method; others used the microtiter plate modification for endpoint quantitation with equal success. For the required products, the relative standard deviation between laboratories (RSD(R)) ranged from 7.4 to 21.6% for 8 fortified (or enriched) products compared with expected (Horwitz equation-based) values of 11-20%. RSD(R) values were higher (22.7-52.9%) for 2 unfortified cereal-grain products. For the optional products, the RSD(R) ranged from 1.8 to 11.2% for 8 fortified products. RSD(R) values were higher (27.9-28.7%) for 2 unfortified cereal-grain products. Based on the results of the collaborative study, the microbiological assay with trienzyme extraction is recommended for adoption as Official First Action.     

Determination of folate in infant formula and adult/pediatric nutritional formula by optical biosensor assay: First Action 2011.05

After a review of data from a single-laboratory validation (SLV) study published in the International Dairy Journal 21, 783-789 (2011), a method for folate in infant formula and adult/pediatric nutritional formula was submitted for consideration of adoption by AOAC as an automated assay that is rapid and simple. The method uses an optical biosensor assay to quantitate total folate content in milk and milk-based pediatric and adult nutritional products. The assay uses folate binding protein and a functionalized sensor surface. The SLV showed an instrumental LOD of 0.1 ng/mL (equivalent to 2.5 microg/100 g for a typical infant formula). The method detection limit was 6.5 microg/100 g with a repeatability of 3.48% and an intermediate reproducibility of 4.63% RSD.     

Dry rehydratable film method for rapid enumeration of coliforms in foods (3M Petrifilm Rapid Coliform Count plate): collaborative study

A rehydratable dry-film plating method for coliforms in foods, the 3M Petrifilm Rapid Coliform Count plate method, was compared with the U.S. Food and Drug Administration's Bacteriological Analytical Manual method for nondairy foods and the American Public Health Association's Standard Methods for the Examination of Dairy Products (SMEDP) method for dairy foods. Six food types, vanilla ice cream, cheddar cheese, fresh refrigerated uncooked pasta, wheat flour, prepared frozen macaroni and cheese, and frozen hash browns, were analyzed for coliforms by 11 collaborating laboratories. For each food product tested, the collaborators received 8 blind samples consisting of a control sample and 3 levels of inoculated sample, each in duplicate. The mean log counts for the methods were comparable. The repeatability and reproducibility variances of the Petrifilm Rapid Coliform Count method at 14 and 24 h were not significantly different from those of the standard methods.     

Optimization and validation of an LC-fLD method for biotin in infant formula, infant cereals, cocoa-malt beverages, and clinical nutrition products

A fast and simple chromatographic method to determine biotin in foods is presented. Biotin is extracted using papain (60 degrees C, 1 h). After pH adjustment and filtration, biotin is determined by LC with fluorescence detection using postcolumn reagent avidin-FITC (avidin labeled with fluorescein isothiocyanate). The method has been validated in a large range of products: milk- and soy-based infant formulas, cereals, cocoa-malt beverages, and clinical nutrition products. The method showed recovery rates of 98.1 +/- 5.7% (average +/- SD) in a large range of concentrations. Biotin concentrations determined in infant formula standard reference materials 1846 and 1849 were in agreement with reference values. RSD of repeatability (RSDr) varied from 2.0 to 4.5%, and intermediate reproducibility (RSD(iR)) from 5.8 to 9.4%. LOD and LOQ were 3.0 and 5.0 microg/100 g, respectively. The proposed method is suitable for routine analysis of biotin in fortified foods (infant formulas, infant cereals, cocoa-malt beverages, and clinical nutrition products). It can be used as a faster, more selective, and precise alternative to the classical microbiological determination, and is easily transferable among laboratories.     

Advanced method using microwaves and solid-phase microextraction coupled with gas chromatography-mass spectrometry for the determination of pyrethroid residues in strawberries

A microwave-assisted desorption method was developed and coupled with solid-phase microextraction and GC-MS for the analysis of pyrethroid residues in strawberries. In the first step, pyrethroid analytes were desorbed from the whole fruits in an aqueous acetonitrile solution at 50% under microwave assistance, so preventing these compounds to be captured with strong matrix effects by endogenous constituents. Then, the 100 microm poly(dimethylsiloxane)-coated fibre was exposed for 30 min in the obtained extracting solution. Calibration curves, realised from blank strawberries spiked at different concentrations with standards, showed a linear range between 1 microg/kg and 250 microg/kg with r2 > 0.992 and variation coefficients below 15%. Limits of detection and quantitation were found lower than 14 microg/kg and 40 microg/kg, respectively. Observed analysis results by using this method and relative to field incurred strawberry samples were also compared to those obtained by two accredited trading laboratories using traditional methods.     

Determination of folate vitamers in food and in Italian reference diet by high-performance liquid chromatography.

A trienzyme treatment (conjugase, alpha-amylase, protease) followed by affinity chromatography and reversed-phase HPLC with UV and fluorescence detection was performed for the quantification of folate vitamers in legumes (chickpea and beans), processed meats (salami Milano and Parma ham) and in an Italian reference diet. This method allowed a good separation of six folate vitamers: 5-methyltetrahydrofolate, 5-formyltetrahydrofolate, folic acid, 10-formylfolic acid, 10-formyldihydrofolate and tetrahydrofolate within 30 min. Recovery, reproducibility and limits of detection of the method are reported. HPLC results were 24-52% lower than the microbiological assay findings.     

A validated liquid chromatographic method for determining folates in vegetables, milk powder, liver, and flour

A liquid chromatographic (LC) method was elaborated for determining folates in foods. Folates were extracted by homogenizing in buffer and heat treatment. A portion was incubated with an enzyme preparation containing conjugase, amylase, and protease. After purification by affinity chromatography, folate monoglutamates were determined by reversed-phase LC with fluorescence and diode array detection. Gradient elution with phosphate buffer and acetonitrile was used to separate vitamers. The most abundant folate forms naturally present in foods were detected, including tetrahydrofolic acid, 5-methyltetrahydrofolic acid, and 5-formyltetrahydrofolic acid. 10-Formylfolic acid could be detected by applying a second fluorescence detector. Folic acid, used for fortification, might also be quantitated with this system. The difference between folate concentrations in sample extracts, with and without treatment of conjugase, is a measure of the quantity of polyglutamates in the food matrixes. An additional treatment with conjugase, amylase, and protease reflects the amount of matrix-bound folates. The LC system gave a linear response over the range 0-100 ng/mL. Detection limit for these compounds were 7 pg/mL for tetrahydrofolic acid and 5-methyltetrahydrofolic acid and 59 pg/mL for 10-formylfolic acid (signal-to-noise ratio > or = 3) when 100 microL was injected. Detection limits for 5-formyltetrahydrofolic acid and folic acid were 1 ng/mL. Repeatability relative standard deviation values for separate folates in 3 candidate Certified Reference Materials (CRMs)--mixed vegetables (CRM 485), pig liver (CRM 487), and whole-meal flour (CRM 121)--and a Certified Reference Material milk powder (CRM 421) varied from 3.3 to 21.0% for the concentration range 1.8-1440 micrograms/100 g. Recoveries ranged from 73 to 109%. Use of amylase and protease was advantageous. Use of a commercially available folate-binding protein for cleanup saved time and money and was effective. Results for 5-methyltetrahydrofolic acid were in good agreement with results obtained with other LC methods. Results for total folates were lower than results obtained with microbiological methods.     

Determination of fortified and endogenous folates in food-based Standard Reference Materials by liquid chromatography-tandem mass spectrometry

The National Institute of Standards and Technology (NIST) is developing a wide variety of Standard Reference Materials (SRMs) to support measurements of vitamins and other nutrients in foods. Previously, NIST has provided SRMs with values assigned for the folate vitamer, folic acid (pteroylglutamic acid), which is fortified in several foods due to its role in prevention of neural tube defects. In order to expand the number of food-based SRMs with values assigned for folic acid, as well as additional endogenous folates, NIST has developed methods that include trienzyme digestion and isotope-dilution liquid chromatography-tandem mass spectrometric (LC-MS/MS) analysis. Sample preparation was optimized for each individual food type, but all samples were analyzed under the same LC-MS/MS conditions. The application of these methods resulted in folic acid values for SRM 1849a Infant/Adult Nutritional Formula and SRM 3233 Fortified Breakfast Cereal of (2.33?±?0.06) μg/g and (16.0?±?0.7) μg/g, respectively. In addition, the endogenous folate vitamer 5-methlytetrahydrofolate (5-MTHF) was detected and quantified in SRM 1849a Infant/Adult Nutritional Formula, candidate SRM 1549a Whole Milk Powder, and candidate SRM 1845a Whole Egg Powder, resulting in values of (0.0839?±?0.0071) μg/g, (0.211?±?0.014) μg/g, and (0.838?±?0.044) μg/g, respectively. SRM 1849a Infant/Adult Nutritional Formula is the first food-based NIST SRM to possess a reference value for 5-MTHF and the first certified reference material to have an assigned 5-MTHF value based on LC-MS/MS. The values obtained for folic acid and 5-MTHF by LC-MS/MS will be incorporated into the final value assignments for all these food-based SRMs.     

Evaluation of VIDAS listeria monocytogenes II (LMO2) immunoassay method for the detection of Listeria monocytogenes in foods: collaborative study

A multilaboratory study was conducted to compare the VIDAS Listeria monocytogenes II (LMO2) immunoassay and the standard cultural methods for the detection of Listeria monocytogenes in foods. Five food types-vanilla ice cream, brie cheese, cooked roast beef, frozen green beans, and frozen tilapia fish-at 3 levels were analyzed by each method. A total of 26 laboratories representing government and industry participated. In this study, 1404 test portions were analyzed of which 1152 were used in the statistical analysis. There were 448 positive by the VIDAS LMO2 assay and 457 positive by the standard culture methods. A chi2 analysis of each of the 5 food types, at the 3 inoculation levels tested, was performed. The resulting chi2 value, 0.36, indicates that overall, there are no statistical differences between the VIDAS LMO2 assay and the standard methods at the 5% level of significance.     

Polycyclic Aromatic Hydrocarbon Burden in Fruit and Vegetable Species Cultivated in Allotments in an Industrial Area

The polycyclic aromatic hydrocarbon (PAH) burden of various fruit and vegetable species (strawberries, apples, tomatoes, lettuce, kohlrabi, potatoes, parsley and kale) cultivated in allotments in the industrial area of Bitterfeld-Wolfen (Germany) were determined. In addition, the garden soils of the sampling sites were analysed. The total PAH concentrations of fruit and vegetables investigated varied in the range of 1-120 µg/kg fresh weight, with the highest being found in parsley and kale. As the highest concentration of benzo(a)pyrene (BaP) was 0.55 µg/kg in a parsley sample, none of the samples exceeded the recommended BaP limit in vegetable foods of 1 µg/kg. A positive relationship between the total PAH burden and the cultivating site was only found for the more highly contaminated species parsley and kale. The total PAH concentrations in the garden soils ranged from 28 to about 7000 µg/kg dry weight.     

Petrifilm rapid S. aureus Count Plate method for rapid enumeration of Staphylococcus aureus in selected foods: collaborative study

A rehydratable dry-film plating method for Staphylococcus aureus in foods, the 3M Petrifilm Rapid S. aureus Count Plate method, was compared with AOAC Official Method 975.55 (Staphylococcus aureus in Foods). Nine foods-instant nonfat dried milk, dry seasoned vegetable coating, frozen hash browns, frozen cooked chicken patty, frozen ground raw pork, shredded cheddar cheese, fresh green beans, pasta filled with beef and cheese, and egg custard-were analyzed for S. aureus by 13 collaborating laboratories. For each food tested, the collaborators received 8 blind test samples consisting of a control sample and 3 levels of inoculated test sample, each in duplicate. The mean log counts for the methods were comparable for pasta filled with beef and cheese; frozen hash browns; cooked chicken patty; egg custard; frozen ground raw pork; and instant nonfat dried milk. The repeatability and reproducibility variances of the Petrifilm Rapid S. aureus Count Plate method were similar to those of the standard method.     

Certification of total mercury and methylmercury concentrations in mussel homogenate (Mytilus edulis) reference material, IAEA-142

Due to the increased demand for new reference materials certified for total and methylmercury (MeHg) a sample of mussel homogenate (IAEA-142) has been prepared. Thirteen experienced laboratories reported results for total Hg of which 9 laboratories also reported results for MeHg content. Laboratories reporting MeHg results used various isolation techniques (solvent extraction, saponification, acid leaching, ion-exchange separation, and distillation) and detection systems (cold vapour atomic absorption spectrometry (CV AAS), cold vapour atomic fluorescence spectrometry (CV AFS), gas chromatography with electron capture detector (GC/ECD) and HPLC with CV AAS detector). In the case of total Hg, most of the laboratories used acid digestion, only two used alkaline dissolution, followed either by CV AAS or CV AFS. One laboratory used neutron activation analyses with radiochemical separation. The data received were in good agreement. The value for total Hg was certified to be 126 ng/g, with a 95% confidence interval from 119 to 132 ng/g. For MeHg the certified value of 47 ng/g expressed as Hg was assigned, with a 95% confidence interval from 43 to 51 ng/g. Stability testing has shown that both total and MeHg are stable if samples are stored in a dry and dark place at room temperature. The sample is now available as a certified reference material and is, in particular, useful for quality control measurements of Hg and MeHg in mussel samples at low concentration levels.     

Certification of total mercury and methylmercury concentrations in mussel homogenate (Mytilus edulis) reference material, IAEA-142

Due to the increased demand for new reference materials certified for total and methylmercury (MeHg) a sample of mussel homogenate (IAEA-142) has been prepared. Thirteen experienced laboratories reported results for total Hg of which 9 laboratories also reported results for MeHg content. Laboratories reporting MeHg results used various isolation techniques (solvent extraction, saponification, acid leaching, ion-exchange separation, and distillation) and detection systems (cold vapour atomic absorption spectrometry (CV AAS), cold vapour atomic fluorescence spectrometry (CV AFS), gas chromatography with electron capture detector (GC/ECD) and HPLC with CV AAS detector). In the case of total Hg, most of the laboratories used acid digestion, only two used alkaline dissolution, followed either by CV AAS or CV AFS. One laboratory used neutron activation analyses with radiochemical separation. The data received were in good agreement. The value for total Hg was certified to be 126 ng/g, with a 95% confidence interval from 119 to 132 ng/g. For MeHg the certified value of 47 ng/g expressed as Hg was assigned, with a 95% confidence interval from 43 to 51 ng/g. Stability testing has shown that both total and MeHg are stable if samples are stored in a dry and dark place at room temperature. The sample is now available as a certified reference material and is, in particular, useful for quality control measurements of Hg and MeHg in mussel samples at low concentration levels. Received: 24 September 1996 / Revised: 20 November 1996 / Accepted: 8 December 1996     

Application of Ultrasound Combined with Acetic Acid and Peracetic Acid: Microbiological and Physicochemical Quality of Strawberries

This work evaluated the application of organic acids (acetic and peracetic acid) and ultrasound as alternative sanitization methods for improving the microbiological and physicochemical qualities of strawberries. A reduction of up to 2.48 log CFU/g aerobic mesophiles and between 0.89 and 1.45 log CFU/g coliforms at 35 °C was found. For molds and yeasts, significant differences occurred with different treatments and storage time (p < 0.05). Ultrasound treatments in combination with peracetic acid and acetic acid allowed a decimal reduction in molds and yeasts (p < 0.05). All evaluated treatments promoted a significant reduction in the Escherichia coli count (p < 0.05). Scanning electron microscopy revealed fragmented E. coli cells due to treatment with acetic acid and ultrasound. Storage time significantly affected pH, total titratable acidity, total soluble solids and the ratio of the total titratable acidity to the total soluble solids (p < 0.05). Anthocyanin content did not change with treatment or time and generally averaged 13.47 mg anthocyanin/100 g of strawberries on fresh matter. Mass loss was not significantly affected by the applied treatments (p > 0.05). The combination of ultrasound and peracetic acid may be an alternative to chlorine-based compounds to ensure microbiological safety without causing significant changes in the physicochemical characteristics of strawberries.     

Determination of vitamin B12 in food products by liquid chromatography/UV detection with immunoaffinity extraction: single-laboratory validation

A fast and simple method to determine vitamin B12 in foods is presented. The method allows, in addition to the determination of added cyanocobalamin, the determination of natural vitamin B12 forms, making it also applicable to nonfortified products, especially those that are milk-based. Vitamin B12 is extracted in sodium acetate buffer in the presence of sodium cyanide (100 degrees C, 30 min). After purification and concentration with an immunoaffinity column, vitamin B12 is determined by liquid chromatography with UV detection (361 nm). The method has been validated in analyses of a large range of products: milk- and soy-based infant formulas, cereals, cocoa beverages, health care products, and polyvitamin premixes. The method showed appropriate performance characteristics: linear response over a large range of concentrations, recovery rates of 100.8 +/- 7.5% (average +/- standard deviation), relative standard deviation of repeatability, RSDr, of 2.1%, and intermediate reproducibility, RSDiR, of 4.3%. Limits of detection and quantitation were 0.10 and 0.30 microg/100 g, respectively, and correlation with the reference microbiological assay was good (R2 = 0.9442). The proposed method is suitable for the routine determination of vitamin B12 in fortified foods, as well as in nonfortified dairy products. It can be used as a faster, more selective, and more precise alternative to the classical microbiological determination.     

Evaluation of proficiency tests in microbiological analysis: enumeration of aerobic microorganisms

South Korea’s Ministry of Food and Drug Safety (MFDS) has been developing programs for the inspection and accreditation of food sanitation inspection institutions. Food sanitation inspection institutions such as MFDS regional offices, the Research Institute of Public Health and Environment and authorized private service providers in South Korea must participate in proficiency testing (PT) programs to comply with the Food Sanitation Act and MFDS Notification No. 2012-112. As the PT provider, the MFDS annually plans various microbiological and chemical PT programs for foods, cosmetics, and pharmaceutical products in accordance with ISO/IEC 17043. The aim of this project was to evaluate the performance of microbiological PT programs to ensure the quality of their routine test results. The test materials used were freeze-dried BioBalls from BTF Pty Ltd. Homogeneity and stability were investigated to assess the adequacy of the selected test materials. This project also contains data from inter-laboratory comparisons organized by MFDS in 2011 and 2012. More than 50 laboratories attended the PT program and submitted their results. Laboratory results were rated with z-scores according to the international standard ISO 13528. The results from 2011 and 2012 revealed that all participating laboratories had similar levels of proficiency. Most of the participants received a rating of “Satisfactory.” Moreover, the percentage of participants who received a rating of “Unsatisfactory” decreased from 3.5 % in 2011 to 2.0 % in 2012.     

Identification and determination of naturally occurring folates in grains of rice (Oryza sativa L.) by UPLC-MS/MS analysis

The genetic potential and biofortification of India-grown rice with bioavailable folate has not been studied yet. The objectives of this study were to determine the folates concentration in four cultivars of rice through UPLC–MS/MS. Total folate concentration in rice cultivars ranged from 11.0 to 51 μg/100 g with a mean of 26.0 μg/100 g. Among the four rice cultivars, the pigmented grain cultivar Nootripathu possesses two-fold rich sources of total folates than the other three non-pigmented grain cultivars. The average value of 100 g serving of rice grains could provide the amount of recommended daily allowance (% RDA) of dietary folates (6.5%) for adults, which ranged from 2.7–12.7%. Among the 5 individual forms of folates, 5-methyltetrahydrofolate was most abundant in rice cultivars followed by 10-Formylfolic acid and folic acid. The result of this study has been useful for biofortification of folates in rice.     

Determination of isoflavones in soy and selected foods containing soy by extraction, saponification, and liquid chromatography: collaborative study.

Isoflavones are biologically active compounds occurring naturally in a variety of plants, with relatively high levels found in soybeans. Twelve laboratories participated in a collaborative study to determine the aglycon isoflavone content of 8 test samples of soy and foods containing soy. The analytical method for the determination of isoflavones incorporates a mild saponification step that reduces the number of analytes measured and permits quantitation versus commercially available, stable reference standards. Test samples were extracted at 65 degrees C with methanol-water (80 + 20), saponified with dilute sodium hydroxide solution, and analyzed by reversed-phase liquid chromatography with UV detection at 260 nm. Isoflavone results were reported as microg/aglycon/g or microg aglycon equivalents/g. The 8 test samples included 2 blind duplicates and 4 single test samples with total isoflavone concentrations ranging from approximately 50 to 3000 microg/g. Test samples of soy ingredients and products made with soy were distributed to collaborators with appropriate reference standards. Collaborators were asked to analyze test samples in duplicate on 2 separate days. The data were analyzed for individual isoflavone components, subtotals of daidzin-daidzein, glycitin-glycitein, and genistin-genistein, and total isoflavones. The relative standard deviation (RSD) for repeatability was 1.8-7.1%, and the RSD for reproducibility was 3.2-16.1% for total isoflavone values of 47-3099 microg/g.     

Stability of penicillin antibiotic residues in meat during storage. Ampicillin

Incurred samples from a pig treated with ampicillin, one of the most important penicillin antibiotic drugs used in food-producing animal treatments, were analyzed at the residue level of the drug in muscle tissue (approximately 100 microg kg(-1)) during their freezing storage and using three different techniques: quantitative microbiological assay, HPLC-UV and LC-MS. Two parameters have been specifically monitored: storage temperature (-20 and -75 degrees C) and storage packaging (ground meat or bulk meat). No significant decrease was observed during the first 3 months of storage monitoring at -20 and -75 degrees C. On the contrary, the sample preparation significantly affected the drug concentration in muscle from the very beginning of the storage. Grinding the meat before storage allowed to keep the drug near the higher level of concentration (approximately 100 microg kg(-1)) when bulk meat stored frozen systematically led to a decreased value (approximately 75 microg kg(-1)). After 8 months of storage at -20 degrees C, a significant decrease arose and was never observed at -75 degrees C. All the results were similarly obtained with the three different techniques used simultaneously, which allows to indicate a good correlation between the techniques.