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目的:建立高效液相色谱法(high performance liquid chromatography,HPLC)测定等量复份膳食中维生素A含量的测定方法。方法:称取10 g样品于平底烧瓶中,加入淀粉酶,混匀,保持45℃30 min。经没食子酸乙醇-氢氧化钠皂化,石油醚提取,40℃通氮气情况下旋转蒸发近干,石油醚溶解、定容,再用氮气吹干,甲醇定容。HPLC法分析,保留时间定性,标准曲线法定量。分析柱为ODS柱(250 mm×46 mm,5μm),流动相:甲醇∶水(98∶2),检测波长325 nm。结果:维生素A在0.25μg/ml～110μg/ml之间线性良好(r>0.9994),平均回收率为97.3%,RSD为1.2%。结论:HPLC法测定等量复份膳食中维生素A的含量简便、快速,灵敏度高,准确性好。
Objective: To establish a method for the determination of vitamin A content in aliquots of duplicate meals by high performance liquid chromatography (HPLC). Methods: Weigh 10 g sample in a flat-bottomed flask, add amylase, mix and maintain 45 ℃ 30 min. The saponification of gallic acid ethanol - sodium hydroxide, extraction of petroleum ether, near 40 ℃ by rotary evaporation near dry, petroleum ether dissolved, fixed volume, and then dried with nitrogen, methanol volume. HPLC analysis, retention time qualitative, standard curve method. The analytical column was ODS column (250 mm × 46 mm, 5 μm). The mobile phase was methanol: water (98: 2) and the detection wavelength was 325 nm. Results: Vitamin A showed a good linearity (r> 0.9994) between 0.25μg / ml and 110μg / ml with an average recovery of 97.3% and a RSD of 1.2%. Conclusion: HPLC method is simple, rapid, sensitive and accurate for the determination of vitamin A content in the duplicate diet.