Kobayashi T, Adachi A, Furuta K
J Nutr Sci Vitaminol (Tokyo). 1976;22(3):215-24. doi: 10.3177/jnsv.22.215.
Gas-liquid chromatographic (GLC) determination of vitamin D3 in tuna liver and vitamin D3 resin oils was investigated and a routine method slightly modified from the previously reported methods (1, 2) was established. Since both tuna liver and vitamin D3 resin oils contained large amounts of sterols, digitonin-Celite column chromatography according to SHEPPARD et al.(4) was used to remove the sterol influence from the unsaponifiable matters of the oils. After collecting the eluate and evaporating the solvent, the residue was subjected to thin-layer chromatography (TLC) using Kieselgel GF254 as an adsorbent and a mixture of n-hexane-ethyl acetate (4: 1) as a developing solvent. The scraped zones corresponding to vitamin D3 and pre-D3 were trimethylsilylated and then applied to the GLC using 1.5% OV-17 packed on Shimalite W (80-100 mesh) as a stationary phase. Trimethylsilylation of the gas chromatograms was an essential procedure, because the peaks of unknown substances in tuna liver oils and lumisterol in vitamin D3 resin oils could not be separated from the peak of pyro-D3 without trimethylsilylation. When the proposed method was applied to the samples, satisfactory results were obtained.
研究了用气液色谱法（GLC）测定金枪鱼肝脏和维生素D3树脂油中维生素D3的含量，并建立了一种在先前报道方法（1,2）基础上稍作修改的常规方法。由于金枪鱼肝脏和维生素D3树脂油中都含有大量甾醇，因此采用了SHEPPARD等人（4）的洋地黄皂苷 - 硅藻土柱色谱法来消除油类不皂化物中甾醇的影响。收集洗脱液并蒸发溶剂后，将残渣用硅胶GF254作为吸附剂，正己烷 - 乙酸乙酯（4:1）混合液作为展开剂进行薄层色谱（TLC）分析。将对应于维生素D3和前维生素D3的刮下区域进行三甲基硅烷化，然后使用填充在Shimalite W（80 - 100目）上的1.5% OV - 17作为固定相进行GLC分析。气相色谱图的三甲基硅烷化是一个必不可少的步骤，因为如果不进行三甲基硅烷化，金枪鱼肝脏油中未知物质的峰以及维生素D3树脂油中麦角钙化醇的峰就无法与焦维生素D3的峰分开。当将所提出的方法应用于样品时，获得了满意的结果。
