高速液体クロマトグラフィーによる栄養機能食品中の水溶性ビタミンの分析

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Rapid and simple methods for the determination of 9 water soluble vitamins in foods with nutrient function claims, so-called dietary supplements, were developed by means of high performance liquid chromatography (HPLG). Calcium pantothenate (PaCa), nicotinic acid (NA), nicotinamide (NAA), thiamine hydrochloride (B_1), riboflavine (B_2) and pyridoxine hydrochloride (B_6-HCl) were extracted from supplements with 0.1% phosphoric acid solution in an 80℃ water bath for 20 min. Folic acid (FA) was extracted with 0.01 N sodium hydroxide solution in an 80℃ water bath for 20 min. These components were separated on an Inertsil ODS-3V column (4.6mm i.d. × 15cm) using a gradient method with 0.1% phosphoric acid solution containing 5mM sodium heptanesulfonate and 0.1% phosphoric acid solution-acetonitrile (70 : 30 v/v) containing 5mM heptanesulfonate. PaCa was detected at UV210nm, NA, NAA, B_1 and B_2 were detected at UV260nm, and B_6-HCl and FA were detected at UV280nm. The recovery of PaCa, NA, NAA, B_1, B_6-HCl and FA (15mg, 7.5mg, 7.5mg, 12.5mg, 5mg and 0.1mg, respectively) added to 3g of supplement was more than 92%. The recovery of B_2 (6mg) from 200mg of the supplement was 99.2%. Cyanocobalamine (B_<12>) was extracted with 0.1% phosphoric acid solution at room temperature, separated on an Inertsil ODS-3V column by using 0.05M potassium dihydrogen phosphate solution-acetonitrile (88 : 12 v/v), and detected at UV360nm. The recovery of B_<12> (30μg) added to 3g of supplement was 94.6%. Dibenzoylthiamine (DBT) was extracted with hydrochloric acid-ethanol-water (8.73 : 700 : 291.27 v/v) solution in an 80℃ water bath for 30 min., degraded with IN sodium hydroxide solution, and thiochrome was produced by oxidation with potassium ferricyanide. Thiochrome was extracted with isobutyl alcohol and separated on an Inertsil ODS-3V column using ethanol-isobutyl alcohol (70 : 30 v/v), and detected with fluorescence detection. The recovery of DBT (200μg) added to 50mg of supplement was 87.3%. Nineteen commercial dietary supplements purchased in the Nara prefecture were determined by the proposed methods. The total number of 9 kinds of water soluble vitamin indicated in these supplements was 119. Sixty one water soluble vitamins in these supplements were indicated the concentration and nutritional function. Fourteen components (23.0%) were less than 80% of the indicated concentrations, and 1 component (1.6%) was more than 180% of the indicated concentration. The disintegration times of these supplements were measured. As a result, one tablet disintegrated over a period greater than 99.9min.
日本食品化学学会の論文
2004-04-27

高速液体クロマトグラフィーによる栄養機能食品中の水溶性ビタミンの分析

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