石岩;郑天骄;魏锋;林瑞超;马双成;
SHI Yan;ZHENG Tian-jiao;WEI Feng;LIN Rui-chao;MA Shuang-cheng;National Institutes for Food and Drug Control;School of Traditional Chinese Materia Medica,Shenyang Pharmaceutical University;Chinese Pharmaceutical University;School of Chinese Materia Medica,Beijing University of Chinese Medicine;

• 1:中国食品药品检定研究院
• 2:沈阳药科大学中药学院
• 3:中国药科大学
• 4:北京中医药大学中药学院

摘要(Abstract)：

针对牛胆粉药材中主要活性组分的化学性质,使用具有较好专属性及准确度的HPLC-ELSD建立可准确反映其内在质量的指纹图谱及主要活性组分含量测定方法。采用Phenomenex Gemini C18(4.6 mm×250 mm,5μm)色谱柱,以10 mmol乙酸铵水溶液与乙腈作为流动相进行梯度洗脱,柱温40℃,ELSD漂移管温度为110℃,ELSD氮气流量为2.8 L·h-1。测得13批牛胆粉的甘氨胆酸、甘氨去氧胆酸、牛磺胆酸和牛磺去氧胆酸的含量,并使用化学计量学对牛胆粉药材指纹图谱进行预处理和模式识别分析。结果样品中甘氨胆酸、甘氨去氧胆酸、牛磺胆酸和牛磺去氧胆酸的平均质量分数分别为(25.2±17.0)%,(4.1±3.4)%,(24.5±20.0)%,(5.2±3.8)%,4者质量分数总和占牛胆粉(59.0±26.0)%。化学计量学分析结果表明自制样品与市售样品差异较大,指纹图谱之中有4处区域的色谱信号较小,但对这种差异的影响却不容忽视。所建HPLCELSD方法简便快速,适合从定性和定量的角度对牛胆粉进行质量评价与控制,可以为完善该药材质控标准提供科学依据。
An HPLC-ELSD method with good specificity and good accuracy was used for the studies of fingerprint and quantification of multi-components for cattle bile powder. The chromatographic analysis was carried out on a Phenomenex Gemini C18column( 4. 6mm × 250 mm,5 μm) with a column temperature of 40 ℃ and a liquid flow-rate of 1. 0 m L·min- 1using 10 mmol ammonium acetate solution and acetonitrile as the mobile phase with a linear gradient. An ELSD was used with a nitrogen flow-rate of 2. 8 L·h- 1,at a drift tube temperature of 110 ℃. The average contents of glycocholic acid,glycodeoxycholic acid,taurocholic acid,taurodeoxycholic acid were( 25. 2 ± 17. 0) %,( 4. 1 ± 3. 4) %,( 24. 5 ± 20. 0) % and( 5. 2 ± 3. 8) % respectively,and the total content of the four bile acids was( 59. 0 ± 26. 0) %. Beyond that,the preprocessing and pattern recognition analysis of the chromatographic fingerprints of samples were applied with chemometric method. The results of this chemometric analysis indicated that the samples from market and self-made samples were different signally,and four regions were noteworthy due to their great impact with poor chromatographic signal.All in one,because this HPLC-ELSD method was simple and accurate,it was suitable for the quality assessment and quality control of cattle bile powder and could be the technological base for its standard perfection.

关键词(KeyWords)： 牛胆粉;胆汁酸;高效液相色谱;蒸发光散射检测器;化学计量学
cattle bile powder;bile acids;HPLC;ELSD;chemometrics

Abstract:

Keywords:

基金项目(Foundation): 国家“重大新药创制”科技重大专项(2014ZX09304307-002)

作者(Author): 石岩;郑天骄;魏锋;林瑞超;马双成;
SHI Yan;ZHENG Tian-jiao;WEI Feng;LIN Rui-chao;MA Shuang-cheng;National Institutes for Food and Drug Control;School of Traditional Chinese Materia Medica,Shenyang Pharmaceutical University;Chinese Pharmaceutical University;School of Chinese Materia Medica,Beijing University of Chinese Medicine;

Email:

DOI:

参考文献(References)：

• [1]南京中医药大学.中药大辞典[M].2版.上海:上海科学技术出版社,2006.
• [2]Qiao X,Ye M,Pan D L,et al.Differentiation of various traditional Chinese medicines derived from animal bile and gallstone:simultaneous determination of bile acids by liquid chromatography coupled with triple quadrupole mass spectrometry[J].J Chromatogr A,2011,1218:107.
• [3]张启明.牛胆粉的化学研究[J].药物生物技术,1997,4(1):58.
• [4]张能荣.牛羊胆汁的质量研究[J].中草药,1983,14(7):15.
• [5]Shoji H,Michiko T,Misako T,et al.Thin-layer chromatography and densitometry of bile components[J].Chem Pharm Bull,1964,12(4):483.
• [6]彭新磊.胆汁、胆酸和胆汁酸[J].生物学通报,1998,33(3):23.
• [7]齐晓慧,那日桑.蒙成药乌力吉-18的临床应用概况[J].中国民族医药杂志,1998,10(3):34.
• [8]中华人民共和国卫生部药典委员会.中华人民共和国卫生部药品标准·蒙药分册[S].呼和浩特:内蒙古科技出版社,1998.
• [9]中国药典.一部[S].2015:7.
• [10]许禄,邵学广.化学计量学方法[M].2版.北京:科学出版社,2004.