SPE-HPLC法检测维生素D滴剂(软胶囊)中的有关物质
Detection of related substances in vitamin D drops(soft capsules)by SPE-HPLC
解植彩 1杨丽娜 1潘寒 1徐庆君 1卢大峰 1李树英 1谢强胜2
作者信息
- 1. 青岛双鲸药业股份有限公司企业技术中心,青岛 266109
- 2. 山东省食品药品检验研究院,济南 250101
- 折叠
摘要
目的:建立固相萃取-高效液相色谱法(SPE-HPLC法)快速检测维生素D滴剂(软胶囊)中维生素D3有关物质反式维生素D3(杂质A)、7-去氢胆固醇(杂质B)、光甾醇3(杂质C)、异速甾醇3(杂质D).方法:取内容物约1.2 g(约相当于维生素D31 548 IU),精密称定,置于试管中,加入异辛烷4 mL,涡旋混匀,采用固相萃取小柱进行净化,以正己烷-乙酸乙酯(85∶15)为洗脱剂,无水乙醇为解吸剂;采用Luna Sil-ica(2)100 Å硅胶柱(150 mm ×4.6 mm,3 μm),以正己烷-正戊醇(996.5∶3.5)为流动相,采用加校正因子主成分自身对照法计算维生素D3各杂质的含量.结果:前维生素D3与植物油分离度＞1.5.维生素D3的线性范围为 0.02～0.80 µg·mL-1,r=0.999 5;杂质 A 的线性范围为 0.02～0.80 μg·mL-1,r=0.999 9;杂质B的线性范围为0.02～0.80 μg·mL-1,r=0.999 9;杂质C的线性范围为0.02～0.80 μg·mL-1,r=0.999 9;杂质D的线性范围为0.02～0.80 μg·mL-1,r=0.999 9.维生素D3杂质A～D的平均回收率(n=9)为93.2％～102.9％.对3批维生素D滴剂(软胶囊)样品进行检测,结果已知杂质及其他最大单个杂质含量均≤0.5％,杂质总含量≤1.0％.结论:本方法经系统方法学验证,适用于维生素D滴剂(软胶囊)中维生素D3有关物质的检测,且具有操作简便、快速、准确的优势,为脂溶性维生素D3制剂的质量控制提供了技术保证.
Abstract
Objective:To establish a solid-phase extraction-high performance liquid chromatography(SPE-HPLC)method for the rapid detection of related substances of vitamin D3,trans vitamin D3(impurity A),7-de-hydrocholesterol(impurity B),lumisterol 3(impurity C),and isotachysterol 3(impurity D),in vitamin D drops(soft capsules).Methods:The content,which was approximately equivalent to vitamin D3 1 548 IU,was taken in about 1.2 g,weighed accurately,put into a test tube,4 mL of isooctane was added to it,and it was swirled and mixed well.Purification was performed using solid phase extraction columns,with n-hexane-ethyl acetate(85∶15)as the elution agent and anhydrous ethanol as the desorption agent.A Luna Silica(2)100 Å silica gel column(150 mm×4.6 mm,3 μm)was used,with n-hexane-n-pentanol(996.5∶3.5)as the mobile phase.The content of vitamin D3 impurities was calculated using the adding standard calibration factors principal compo-nent self-control method.Results:The separation degree between pre-vitamin D3 and vegetable oil was greater than 1.5.The linear range of vitamin D3 was 0.02-0.80 μg·mL-1,with r=0.999 5.The linear range of im-purity A was 0.02-0.80 μg·mL-1,with r=0.999 9.The linear range of impurity B was 0.02-0.80μg·mL-1,with r=0.999 9.The linear range of impurity C was 0.02-0.80 μg·mL-1,with r=0.999 9.The linear range of impurity D was 0.02-0.80 μg·mL-1,with r=0.999 9.The average recovery rate of impurities A-D(n=9)was 93.2％-102.9％.The detection results of 3 batches of vitamin D drops(soft capsules)showed that the content of known impurities and other maximum single impurities were less than 0.5％,and the total content of impurities was less than 1.0％.Conclusion:The results indicate that the established method is suitable for the detection of vitamin D3 related substances in vitamin D drops(soft capsules).It has the advanta-ges of being easy to operate,providing rapid and accurate results,and providing technical assurance for the quali-ty control of fat-soluble vitamin D3 preparations.
关键词
固相萃取/高效液相色谱法/维生素D3有关物质/工艺杂质/降解杂质Key words
solid phase extraction/high performance liquid chromatography/vitamin D3-related substances/process impurities/degradation impurities引用本文复制引用
出版年
2024
NETL
NSTL
万方数据