RP-HPLC同时测定保健食品中维生素D_2和D_3的含量

Determination of Vitamin D_2 and Vitamin D_3 in Health Food by RP-HPLC

建立反向高效液相色谱法（RP-HPLC）同时测定保健食品中维生素D_2和D_3的检验方法。片剂和硬胶囊剂采用超声提取,软胶囊采用皂化提取制备,Zorbak ODS色谱柱（4.6 mm×250 mm×5μm）,以甲醇-乙腈（9∶1,V/V）为等度流动相,检测波长为264 nm,流速为1.0 m L/min,柱温35℃。结果表明,维生素D_2在0.07-7.14μg/m L质量浓度范围、维生素D_3在0.08-8.62μg/m L质量浓度范围内呈线性关系（r=0.999 5）,维生素D_2和维生素D_3的回收率分别为83.49%-98.56%和83.71%-98.43%,定量限为0.45μg/100 g。经方法学验证符合相关标准。该方法简便快捷,准确度高,能满足含维生素D类保健食品的含量测定要求。

Simultaneous determination of vitamin D2 and D3 in health food was used by reversed phase high performance liquid chromatography. The tablet and hard capsule were extracted by ultrasonic extraction, soft capsule was extracted by saponification, Zorbak ODS chromatographic column （4.6 mm×250 mm×5 μm） was used, with the mobile phase of methanol and acetonitrile （9 ： 1, V/V）, detection was at 264 nm, the flow rate was 1.0 mL/min, the column temperature RPO was 35 ℃. The results showed that the calibration curve of vitamin D2 was linear in the range of 0.07 to7.14 μg/mL, which the vitamin D3 was linear in the range of 0.08 to 8.62 μg/mL （r=0.999 5）, recoveries of vitamin D2 and vitamin D3 were 83.49%-98.56% and 83.71%-98.43%, limit of quantification was 0.45μg/100 g. After validation, the method was in accordance with the relevant standards. The method was simple, rapid and accurate, and could meet the requirement of the content determination of vitamin D in health foods.