Tsukida K, Kodama A, Saiki K
J Nutr Sci Vitaminol (Tokyo). 1976;22(1):15-20. doi: 10.3177/jnsv.22.15.
The simultaneous determination of vitamin D2 and its isomers by high-speed liquid chromatography (HSLC) is described. Preferred operating conditions for the resolution and quantitation of vitamin D2 5,6-trans-vitamin D2, ergosterol, lumisterol2, isovitamin D2, isotachysterol2, tachysterol2, and previtamin D2 are as follows: column, "Zorbax" SIL (25 cm x 2.1 mm i.d.); pressure, 100-120 kg/cm2; temperature, ambient; detector, UV 254 nm; mobile phase, 0.15% methanol + 2% ether in pentane (for the simultaneous determination), 10% ether in hexane (for quantitation of the practical sets of isomers), or 55% CHCl3 (distilled) in pentane (for separation of isovitamin D2 and 5,6-trans-vitamin D2); sample size, 1 mul (greater than 1-5 ng); internal standard, p-cresol or alpha-naphthol. High-speed, good resolution, precise and accurate trace analysis, greater analysis flexibility, no necessity for preparing derivatives, and gentle operating conditions are main potential advantages of our method which has proved to be very efficient and surpasses all analytical procedures hitherto proposed.
本文描述了用高效液相色谱法（HSLC）同时测定维生素D2及其异构体的方法。维生素D2、5,6-反式维生素D2、麦角固醇、光甾醇2、异维生素D2、异速甾醇2、速甾醇2和前维生素D2分离与定量的优选操作条件如下：色谱柱，“Zorbax”SIL（25 cm×2.1 mm内径）；压力，100 - 120 kg/cm²；温度，室温；检测器，紫外254 nm；流动相，0.15%甲醇 + 2%乙醚的戊烷溶液（用于同时测定）、10%乙醚的己烷溶液（用于实际异构体组的定量）或55%（蒸馏）氯仿的戊烷溶液（用于分离异维生素D2和5,6-反式维生素D2）；进样量，1 μl（大于1 - 5 ng）；内标，对甲酚或α-萘酚。高速、高分辨率、精确准确的痕量分析、更大的分析灵活性、无需制备衍生物以及温和的操作条件是我们方法的主要潜在优势，该方法已被证明非常有效，且优于迄今提出的所有分析方法。
