生物发酵骨化二醇相关杂质的制备及结构鉴定

The Preparation and Identification of the Related Impurities from Biotransfomated Calcifediol

摘要:

目的 对生物发酵的骨化二醇相关杂质进行分离纯化及结构鉴定。方法 生物发酵维生素D3的发酵液经过粗提后，采用高压制备色谱多次分离得到骨化二醇的相关杂质，使其达到一定纯度后，再通过DAD扫描、质谱和核磁鉴定化合物的结构。结果 纯化后得到纯度97.5%的杂质单品，结构鉴定此杂质化学结构式为(5Z,7E)-9,10-开环胆甾-5,7,10(19)-三烯-3b,24-二醇，也即24-羟基维生素D3，为骨化二醇的同分异构体。结论 本研究结果为生物发酵的骨化二醇原料药的质量标准的建立提供依据，同时为后续实验工作提供杂质对照品.

Abstract:

OBJECTIVE To isolate, purify and identify the structures of related impurities from biotransfomated calcifediol. METHODS After vitamin D3爁ermentation broth was crudely extracted, high-pressure preparative chromatography was adopted to isolate calcifediol repeatedly, acquiring high purity related impurities from calcifediol. When the impurities reached a certain purity, through DAD Scanning, Mass Spectrometry and Nuclear Magnetic Resonance, the structure of chemical compound was identified. RESULTS After purification, 97.5% purity of impurity item was obtained. After structure identification, the impurity is (5Z,7E)-9,10-ring opening 5,7,10(19)-triene-3b,24-diol which means that 24-hydroxy vitamin D3 is the isomer of calcifediol. CONCLUSION The results of this study provide the basis for the establishment of quality standard of biotransfomated calcifediol as active pharmaceutical ingredients. Meanwhile, they also provide impurity reference for subsequent experiental work.