高效液相色谱-串联三重四极杆质谱法测定维生素D/AD滴剂(软胶囊)中维生素D_(3)的含量

Determination of vitamin D_(3)in vitamin D/AD drops(soft capsules)by high performance liquid chromatography-tandem triple quadrupole mass spectrometry

目的:建立简单快速的高效液相色谱-串联三重四极杆质谱法测定维生素D/AD滴剂(软胶囊)中维生素D_(3)含量。方法:取制剂内容物适量,直接用正己烷稀释至合适浓度后进行检测;采用Agilent Zorbax Rx-Sil(100 mm×3.0 mm,1.8μm)色谱柱,流动相为正己烷-正戊醇(997∶3),流速1 mL·min^(-1),柱温35℃,大气压化学电离(APCI)源,MRM模式,m/z 385.3→259.1(+)(定量离子),流路0~5 min切废液,5 min后进质谱。结果:前维生素D_(3)与维生素D_(3)在质谱检测中有相同的响应因子,可直接合并峰面积进行含量计算;维生素D_(3)总量的线性范围0.01~0.5μg·mL^(-1),r=0.9995;4种维生素D/AD滴剂(软胶囊)样品中维生素D_(3)平均回收率(n=9)在100.5%~102.3%。对4种维生素D/AD滴剂(软胶囊)进行多批次测定,有效期内的制剂样品中的维生素D_(3)均达到标示量的90.0%~105.0%。结论:本法经方法学验证,适用于维生素D/AD滴剂(软胶囊)中的维生素D_(3)含量测定,无需任何前处理步骤,无需测定前维生素D_(3)的相对校正因子,操作简便、快速准确,为其他油溶性维生素D_(3)制剂的含量检测提供参考。

Objective:To establish a simple and rapid method to determine the content of vitamin D_(3)in vitamin D/AD drops(soft capsules).Methods:The appropriate amount of the content of the preparation was directly diluted with n-hexane to a suitable concentration for high performance liquid chromatography-tandem triplequadrupole mass spectrometry(LC-QQQ)detection.The chromatographic columnwas Agilent Zorbax Rx-Sil(100 mm×3.0 mm,1.8μm).The mobile phase was n-hexane-n-pentane alcohol(997∶3).Flow rate:1 mL·min^(-1),column temperature:35℃,APCI source,MRM mode,m/z 385.3→259.1(+)(quantitative ion).0-5 min to waste,and then after 5 min to mass.Results:The pre-vitamin D_(3)and vitamin D_(3)had the same response factor in the mass spectrometry detection,and their peak area could be directly combined to calculate the content;the linear range of total vitamin D_(3)was 0.01-0.5μg·mL^(-1)(r=0.9995);the average recoveries(n=9)of vitamin D_(3)in the four vitamin D/AD drops(soft capsules)samples were between 100.5%and 102.3%.Multi-batch of the four vitamin D/AD drops(soft capsules)were detected,and the vitamin D_(3)in the preparation samples were within the validity period reached 90.0%-105.0%of the labeled amount.Conclusion:This method has been validated by methodology and is suitable for the determination of vitamin D_(3)content in vitamin D/AD drops(soft capsules)without any pre-treatment steps or determination of the relative correction factor of the precholecalciferol.The operation is simple,fast and accurate,and provides a reference for the content detection of other oil-soluble vitamin D_(3)preparations.