二维液相色谱法测定维生素D2软胶囊中速甾醇的含量

Determination of the Content of Tachysterol in Vitamin D2 Soft Capsules by Two-dimensional Liquid Chromatography

关键词:  二维液相色谱法  速甾醇  维生素D₂软胶囊  主成分自身对照法

Key words:  two-dimensional liquid chromatography  tachysterol  vitamin D₂ soft capsules  self-principal component contrast method

摘要:

目的 建立二维液相色谱法检测维生素D₂软胶囊中速甾醇含量的方法。方法 检测波长281 nm；流速0.5 mL·min^-1；柱温40℃；一维色谱条件：采用Thermo Acclaim HILIC丙基酰胺键合硅胶柱（2.1 mm×150 mm，3 μm），以正己烷-正戊醇-异丙醇（998：0.5：1.5）为流动相；二维色谱条件：采用Thermo硅胶柱（3 mm×100 mm，1.8 μm），以正己烷（A）-正己烷-正戊醇-异丙醇（96：2：2）（B）为流动相，梯度洗脱。结果 速甾醇在0.032 5~0.325 μg·mL^-1内与峰面积的线性关系良好，r=0.999 9；回收率为99.35%（RSD=1.4%，n=9），速甾醇的相对校正因子为0.047 6，检测限为1.6 ng·mL^-1，定量限为4.8 ng·mL^-1。结论 建立的方法符合杂质测定的要求，可以准确测定维生素D₂软胶囊中速甾醇的含量。

Abstract:

OBJECTIVE To establish a method for the determination of tachysterol in vitamin D₂ soft capsules using two-dimensional liquid chromatography. METHODS Test wavelength: 281 nm; velocity of flow 0.5 mL·min^-1; column temperature: 40 ℃. One-dimensional chromatographic condition: Thermo Acclaim HILIC propylamide bonded silica gel column (2.1 mm×150 mm, 3 μm) was used, hexane-pentyl alcohol-isopropanol(998：0.5：1.5) as mobile phase. Two-dimensional chromatographic condition: Thermo silica column(3 mm×100 mm, 1.8 μm) was used, hexane(A)-hexane-pentyl alcohol- isopropanol(96：2：2)(B) as mobile phase, gradient elution. RESULTS In the range of 0.032 5-0.325 μg·mL^-1, tachysterol could obtain a good linear relationship(r=0.999 9), the recovery of tachysterol was 99.35%(RSD=1.4%, n=9). The relation correction factor of tachysterol was 0.047 6, the LOD was 1.6 ng·mL^-1, the LOQ was 4.8 ng·mL^-1. CONCLUSION The method conform to the requirement of impurity determination, and the content of the tachysterol in vitamin D₂ soft capsules can be accurately measured.