Determination of vitamin B12 in dry feeds by atomic absorption spectrophotometry.

Vitamin B12 was determined in dry feeds by atomic absorption spectrophotometry (AAS). Samples containing B12 were extracted with an assay solution, 5 g EDTA was added to the filtrate, the pH was adjusted to 7 with NH4OH, and 5 g charcoal was added. The charcoal was removed by filtering through ashless paper which was then placed in a beaker and ashed at 600 degrees C. After dissolving the cobalt oxide from the ash in 5N HNO3, cobalt content was determined by using AAS. To determine mg B12/lb feed, ppm cobalt in the feed is multiplied by 10.43. The sensitivity of the proposed procedure is 1 mg vitamin B12/lb. The procedure is rapid and precise, and results compare favorably with AOAC method 43.109.     

Disposable sensor for measurement of vitamin B(2) in nutritional premix, cereal, and milk powder

Methodologies for the determination of vitamin B(2) in food matrixes and a premix using simple sample conditioning steps coupled with a convenient and cheap electrochemical sensing device are presented. Electrochemical analysis based on differential pulse voltammetry (DPV) coupled to carbon electrodes gave a well-defined reduction peak at -0.42 V versus a Ag/AgCl quasi-reference electrode. Using a straightforward sample preparation step, vitamin B(2) can be measured successfully in a nutritional premix and food products. Standard additions of riboflavin were used to confirm the analyte concentrations and to provide precision data.     

Determination of vitamin B(6) in cooked sausages

A reverse-phase high-performance liquid chromatography (HPLC) method has been described for the determination of various active forms of vitamin B(6) in meat products. Different extracting agents were tested to solubilize fully the analyte for quantification. The best data were obtained by extracting the samples with 5% (w/v) metaphosphoric acid. Separation by HPLC was performed with fluorescence detection (excitation, 290 nm; emission, 395 nm), on a 10 cm x 0.46 cm i.d. Hypersil BDS C(18) 5 microm column using a mixture of 50 mM phosphate buffer (pH 3.2) and acetonitrile (99:1, v/v) as mobile phase. Precision of the method was 0.5% (within a day) and 4.3% (between days). The detection limits were 0.020 mg/100 g for pyridoxal and pyridoxamine, 0.017 mg/100 g for pyridoxamine phosphate, 0.500 mg/100 g for pyridoxal phosphate, and 0.033 mg/100 g for pyridoxol, with a signal-to-noise ratio of 3. The recovery ranged from 92.0 to 100.0%.     

Glutamine in commercial liquid nutritional products

Total glutamine concentrations in commercial nutritional products have been determined by enzymatic hydrolysis followed by HPLC quantification of free glutamine and free pyroglutamic acid. Hydrolysis was accomplished by a published three-enzyme (Pronase, leucine aminopeptidase, prolidase), 20-h/37 degrees C digestion. Glutamine was determined as its FMOC derivative by reverse phase HPLC-fluorescence, and pyroglutamic acid was determined directly by organic acid HPLC-UV. Approximately 4.11% of the released glutamine is converted to pyroglutamic acid during the 20-h digestion. Experimental ratios of enzyme hydrolysis glutamine to acid hydrolysis glutamic acid + glutamine + pyroglutamic acid (GLX) indicate that the method recovers >90% of the protein-bound glutamine. The nutritional products with casein dominant intact protein systems typically deliver >9 g of glutamine/100 g of protein, or approximately 40 g of glutamine/100 g of GLX.     

Determination of gibberellic acid in fermentation broth and commercial products by micellar electrokinetic chromatography

Micellar electrokinetic chromatography (MEKC) was developed as a method for quantitative determination of gibberellic acid (GA3) in fermentation broth and commercial products, using 25 mM disodium tetraborate as a buffer at pH 9.2 and 100 mM sodium dodecyl sulfate as a micellar phase. The baseline resolution (Rs of GA3 from other compounds in fermentation broth was achieved with Rs > 2.5. The addition of methanol or acetonitrile in the MEKC buffer did not give a better resolution. Advantages of this MEKC method include high accuracy and precision and no sample preparation except for dilution and filtration.     

Differential pulse polarographic determination of iodine in foods and nutritional products

A differential pulse polarographic procedure is described for the analysis of iodine in foods and nutritional products. Samples with iodine concentrations ranging from 5000 to 0.2 ppm have been successfully analyzed using this procedure. Precision averaged between 2 and 10% relative to the iodine level measured. Recoveries of added iodine ranged from 89 to 108% with external standards, and 97-100% by an analyte additions technique. Samples analyzed include dried almonds, whey protein concentrate, nonfat dried milk, sea kelp, vitamin-mineral nutritional supplements, diet meal replacement products, dried green beans, dried mushrooms, and wheat germ.     

Rapid determination of polyphenols and vitamin C in plant-derived products

Polyphenols, widely spread in our diet by the consumption of plant food products, are commonly determined using Folin-Ciocalteu reagent that interacts with other different reducing nonphenolic substances and leads to an overestimation of polyphenol content. In this paper we report an optimized Folin-Ciocalteu method to specifically determine the contents of total polyphenols and vitamin C. After the optimal conditions for the colorimetric assay were set, solid-phase extraction (Oasis HLB (hydrophilic-lipophilic balance)) was carried out to eliminate the water-soluble reducing interferences including vitamin C. Colorimetric correction was thus performed by subtracting interfering substances contained in the water washing extract from the raw extract. Moreover, vitamin C present in the water washing extract can be destroyed by heating and thus colorimetrically deduced. This procedure was set up with synthetic solutions and validated on different extracts from fruit products.     

Maternal serum vitamin B12, folate and homocysteine and the risk of neural tube defects in the offspring in a high-risk area of China

OBJECTIVE: To examine the association between the risk of neural tube defects (NTD) and maternal serum vitamin B12, folate and homocysteine in a high-risk area of China. DESIGN: A case-control study was carried out in Luliang mountain area of Shanxi Province.Subjects/settingA total of eighty-four NTD pregnancies and 110 matched controls were included in the study; their serum vitamin B12 and folate concentrations were measured by chemiluminescent immunoenzyme assay and total homocysteine concentrations by fluorescent polarisation immunoassay. RESULTS: Serum vitamin B12 and folate concentrations were lower in NTD-affected pregnant women than in controls (P < 0.01). Serum total homocysteine was higher in the NTD group than in controls at less than 21 weeks of gestation (P < 0.01). Adjusted odds ratios revealed that women with lower vitamin B12 (adjusted OR=4.96; 95 % CI 1.94, 12.67) and folate (adjusted OR=3.23; 95 % CI 1.33, 7.85) concentrations had a higher risk of NTD compared to controls. Based on dietary analysis, less consumption of meat, egg or milk, fresh vegetables and fruit intake would increase the risk of NTD. CONCLUSIONS: Lower serum concentrations of folate and vitamin B12 are related to the increased risk of NTD in high-risk populations. Both folate and vitamin B12 intake insufficiency could contribute to the increased risk of NTD. A dietary supplement, combining folate and vitamin B12, might be an effective measure to decrease the NTD incidence in these areas.     

Determination of sterol and fatty acid compositions, oxidative stability, and nutritional value of six walnut (Juglans regia L.) cultivars grown in Portugal

Six cultivars (Franquette, Marbot, Mayette, Mellanaise, Lara, and Parisienne) of walnuts (Juglans regia L.) were collected during the 2001 crop, from Bragan?a, Portugal. Chemical composition, including moisture, total oil content, crude protein, ash, carbohydrates, and nutritional value, was evaluated. Fat was the predominant component, ranging from 62.3 to 66.5%. Total oil was extracted and analyzed for fatty acids, sterols, oxidative stability, and peroxide value. Fatty acids and sterols were determined by gas-liquid chromatography coupled to a flame ionization detector. Eighteen fatty acids were quantified. Polyunsaturated fatty acids and, in particular, linoleic acid were predominant. Beta-Sitosterol, delta(5)-avenasterol, and campesterol were the major sterols found. Differences were observed among the studied cultivars, especially in peroxide values and in the sterol profile.     

Certification of B-group vitamins (B1, B2, B6, and B12) in four food reference materials

In 1989, the Community Bureau of Reference started a research program to improve the quality of vitamin analysis in food. To achieve this task, vitamin methodology was evaluated and tested by interlaboratory studies and the preparation of certified reference materials, which will be used for quality control of vitamin measurements. The main improvements in methodology were achieved by testing and standardizing the extraction condition and enzymatic hydrolysis procedures. Results for each individual material are derived from five replicate determinations using at least two independent methods: liquid chromatography (HPLC) and microbiological assay for vitamins B1, B2, and B6; and radioprotein binding and microbiological assays for vitamin B12. The certificate of analysis for four reference materials gives mass fraction values for water-soluble vitamins. These certified values were based on the acceptable statistical agreement of results from collaborating laboratories. Certified values with uncertainties (mg/kg dry matter) for each CRM are as follows: 4.63 (0.20) and 4.10 (0.51) for vitamins B1 and B6, respectively, in CRM 121 (wholemeal flour); 6.51 (0.24), 14.54 (0.3), 6.66 (0.43), and 0.034 (0.003) for vitamins B1, B2, B6, and B12, respectively, in CRM 421 (milk powder); 3.07 (0.17) and 4.80 (0.40) for vitamins B1 and B6, respectively, in CRM 485 (lyophilized mixed vegetables), and 8.58 (0.55), 106.8 (2.8), 19.3 1.5), and 1.12 (0.044) for vitamins B1. B2, B6, and B12, respectively, in CRM 487 (lyophilized pig liver).     

Dietary intake of vitamin B6 and concentration of vitamin B6 in blood samples of German vegans

OBJECTIVE: The study aimed to evaluate the dietary vitamin B6 intake and determine the vitamin B6 concentration in blood samples of German vegans. DESIGN AND SETTING: Cross-sectional study with 33 examination sites all over Germany.Subjects Ninety-three vegans (50 females) with a mean (+/- standard deviation (SD)) age of 43.7 +/- 15.7 years who took no vitamin supplements. METHODS: Dietary intake was assed using a semi-quantitative food-frequency questionnaire. Erythrocyte aspartate aminotransferase activity coefficient (EAST-AC) was calculated as the ratio of stimulated (pyridoxal 5'-phosphate added) to unstimulated activity in blood samples that were provided after an overnight fast. RESULTS: Mean +/- SD vitamin B6 intake was 2.83 +/- 0.98 mg day(-1) and mean +/- SD protein intake was 56.6 +/- 21.7 g day(-1). Of the participants 4% showed vitamin B6 intakes lower than daily recommended intakes for Germany, 16% showed EAST-AC > 1.85, and a further 58% showed EAST-AC of 1.5-1-85. Moderate vegans were affected to a lesser extent than strict vegans. None of the established confounders was a significant predictor of EAST-AC. In logistic regression analyses the contribution of nutriments and cereals to pyridoxine intake was the only predictor of EAST-AC classified as < or = 1.85 and > 1.85, respectively. CONCLUSIONS: In spite of the high total intake of vitamin B6, an adequate concentration in blood samples could not be realised for a majority of the participants. Due to the health implications of a marginal pyridoxine status, vegans should be encouraged to include foods with a high bioavailability of pyridoxine, such as beans, lentils and bananas, in the daily diet.     

Characterization of a vitamin B12 compound from unicellular coccolithophorid alga (Pleurochrysis carterae).

A unicellular coccolithophorid alga, Pleurochrysis carterae, contained 125.4 +/- 1.2 microg of vitamin B12 per 100 g dry cell weight of the lyophilized algal cells. A vitamin B12 compound was purified from the lyophilized algal cells and partially characterized. The silica gel 60 TLC and reversed-phase HPLC patterns of the purified pink-colored compound were identical to those of authentic vitamin B12, but not those of vitamin B12 analogues inactive for humans. When 22-week-old B12-deficient rats which excreted substantial amounts of methylmalonic acid (75.5 +/- 12.3 mg/day) in urine were fed the P. carterae (10 g per kg diet)-supplemented diet for 12 d, urinary methylmalonic acid excretion (as an index of vitamin B12 deficiency) of the rats became undetectable and hepatic vitamin B12 level of the rats was significantly increased.     

Automated analysis of vitamin C in pharmaceutical products.

The determination of total vitamin C in the form of both l-ascorbic acid (AA) and dehydroascorbic acid (DHAA) present in pharmaceutical preparations has been automated. Total vitamin C (completely oxidized to DHAA) was determined by reaction with 2,4-dinitrophenylhydrazine while blanks utilized the same reagent after reducing all DHAA to AA. The automated method was applicable to a variety of multivitamin preparations including those containing iron and copper. The mean recovery of L-ascorbic acid added to 11 multivitamin preparations was 99.4% with a coefficient of variation of 2.5%. In the analysis of these products, results obtained by the automated method were essentially the same as those obtained by the original manual method. For preparations containing no copper salts, the results were also comparable to those obtainable by titration with 2,6-dichloroindophenol except in 1 product which contained some DHAA.     

Titrimetric determination of vitamin B6 using chloramine-T, bromamine-T, and bromamine-B

Simple titrimetric methods have been developed for determination of the vitamin pyridoxine (PRX) in solution, using aromatic N-haloamines, chloramine-T, bromamine-T, and bromamine-B. The method is based on oxidation of PRX by the N-haloamines; the oxidation involves a 2-electron change. The reaction products, pyridoxal and sulfonamides, were identified. The effects of variables, such as pH of the medium and the presence of foreign ions, and pyridoxal, pyridoxalamine, glycerol, and alcohol, on the rate of oxidation were studied. The methods are useful for determining PRX in pharmaceutical formulations and for assaying its cadmium complex. Statistical evaluation showed that analytical results are accurate to within 1%.