提出了用反相高效液相色谱法(RP-HPLC)分离和测定胶囊、油状试剂中维生素D2及维生素D3的含量。样品用甲醇超声提取,提取液用甲醇定容至100 mL,经0.45 μm滤膜过滤后供HPLC分析。采用Zorbax SB-C18反相色谱柱及C18保护柱作为分离柱,乙腈作为流动相,流量为1.0 mL·min-1,在波长265 nm处作紫外检测,进样量为20 μL。维生素D2和维生素D3的质量浓度在0.50~20 mg·L-1范围内与相应的峰面积呈线性关系,测得检出限(3S/N)依次为0.030,0.026 mg·L-1。此方法应用于实样分析并在此基础用标准加入法做回收试验,同时进行精密度试验,测得回收率在95.0%~99.4%之间,测定值的相对标准偏差(n=6)在1.2%~2.6%之间。

Vitamin D2 and D3 either in capsules or in oil preparations were determined by RP-HPLC. The sample was extracted ultrasonically with methanol. The extract was diluted to 100 mL with methanol, which was used for HPLC analysis after filtering on 0.45 μm filtering membrane. The reverse phase chromatographic column Zorbax SB-C18 and the protective column C18 were used for separation with pure acetonitrile as mobile phase (flow rate 1.0 mL·min-1). UV detection at the wavelength of 265 nm was adopted in the determination. Volume of sample solution taken for introduction was 20 μL. Linear relationships between values of peak area and mass concentration of both Vitamin D2 and D3 were found in the same range of 0.50-20 mg·L-1. Detection limit (3S/N) of 0.030 mg·L-1 was found for Vitamin D2 and 0.026 mg·L-1 for Vitamin D3. This method was used in analysis of substancial samples and using these samples as matrixes, tests for recovery were made by standard addition method, values of recovery were found in the range of 95.0%-99.4%, with values of RSD′s (n=6) in the range of 1.2%-2.6%.