目的 应用高效液相色谱-离子阱质谱联用技术(HPLC-MS/MS)对熊胆粉、熊胆粉中间体及痰热清胶囊中胆汁酸类成分进行定性分析。方法 以体积分数70%甲醇作为提取溶剂制备供试液,色谱分离选用Ultimate XB C18色谱柱(4.6 mm×250 mm, 5 μm),以乙腈-0.1%甲酸水为流动相进行梯度洗脱,电喷雾离子阱质谱正负离子模式下采集数据,根据化合物的二级质谱裂解碎片和色谱峰的保留时间等信息结合对照品的碎片裂解规律和文献数据进行结构鉴定。结果 从熊胆粉、中间体和胶囊中共鉴定或推测33个化合物,其中牛磺熊去氧胆酸、牛磺鹅去氧胆酸、熊去氧胆酸、鹅去氧胆酸、7α-羟基-3-氧代-5β-胆烷酸以及一未知成分共6种化学成分在痰热清胶囊生产过程中从熊胆粉经由熊胆粉中间体最终转移至痰热清胶囊中。熊胆粉中间体制备过程中新生成成分有21个,主要为游离型胆汁酸,其中19个化合物均转移至胶囊中。结论 通过痰热清胶囊生产过程中熊胆粉化学成分变化情况进行考察,为痰热清胶囊生产过程的质量控制与评价提供了参考依据。
Abstract
OBJECTIVE To identify the chemical components of bile acids from bear bile powder, the intermediate of bear bile powder and Tanreqing capsules by HPLC-MS/MS. METHODS All samples were extracted with 70% methanol(V/V), and an Ultimate XB C18 column (4.6 mm×250 mm, 5 μm) was employed for separation with acetonitrile-0.1% formic acid as mobile phase in gradient elution. The MS spectrum was acquired in both positive and negative ion mode using ESI ion source. The chemical components were identified by the second mass spectrometric pyrolysis fragments, chromatographic peak retention time and fragmentation regularity summarized from the reference standards and the available literature. RESULTS A total of 33 compounds were successfully identified or tentatively predicted, and six chemical compounds including tauroursodeoxycholic acid, taurochenodeoxycholic acid, ursodeoxycholic acid, chenodeoxycholic acid, 7α-hydroxy-3-oxo-5β-cholanic acid and one unknown constituent were finally transferred to Tanreqing capsules through the intermediate of bear bile powder. Moreover, 21 new chemical compounds (major ingredients of free bile acids) were generated during the production process of the intermediate, and 19 components were also detected in Tanreqing capsules. CONCLUSION The investigation of the change of constituents in bear bile powder during Tanreqing capsules production provides a basis for the quality control and evaluation of Tanreqing capsules during production process.
关键词
熊胆粉 /
痰热清胶囊 /
胆汁酸类 /
高效液相色谱-离子阱质谱联用 /
生产过程
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Key words
bear bile powder /
Tanreqing capsules /
bile acid /
HPLC-MS/MS /
production process
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中图分类号：
R917
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参考文献
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脚注
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基金
国家自然科学基金资助项目(81403089)
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