高效液相色谱-电雾式检测器测定牛黄解毒片中的6种胆汁酸

引用本文: 林楚慧, 柴瑞平, 张璐, 付志博, 章弘扬, 张敏, 胡坪. 高效液相色谱-电雾式检测器测定牛黄解毒片中的6种胆汁酸[J]. 分析化学, 2022, 50(5): 764-771. doi: 10.19756/j.issn.0253-3820.210910 shu

Citation: LIN Chu-Hui, CHAI Rui-Ping, ZHANG Lu, FU Zhi-Bo, ZHANG Hong-Yang, ZHANG Min, HU Ping. Determination of Six Kinds of Bile Acids in NiuhuangJiedu Tablets Using High Performance Liquid Chromatography Coupled with Charged Aerosol Detector[J]. Chinese Journal of Analytical Chemistry, 2022, 50(5): 764-771. doi: 10.19756/j.issn.0253-3820.210910 shu

高效液相色谱-电雾式检测器测定牛黄解毒片中的6种胆汁酸

林楚慧 ^1, ,
柴瑞平 ^2, ,
张璐 ^1, ,
付志博 ^1, ,
章弘扬 ^1, ,
张敏 ^3, ,
胡坪 ^{1,
,}

1.
华东理工大学化学与分子工程学院, 上海市功能性材料化学重点实验室, 上海 200237;
2.
赛默飞世尔科技中国有限公司, 上海 200000;
3.
华东理工大学药学院, 上海市新药设计重点实验室, 上海 200237

通讯作者: 胡坪,E-mail:huping@ecust.edu.cn

基金项目:
国家自然科学基金项目（No.81973285）资助。

摘要: 建立了牛黄解毒片中6种胆汁酸类物质（甘氨胆酸、猪脱氧胆酸、胆酸、甘氨脱氧胆酸、鹅脱氧胆酸与脱氧胆酸）的高效液相色谱-电雾式检测器（HPLC-CAD）测定方法。样品经70%（V/V）甲醇超声提取，氮吹浓缩后，以Agilent Poroshell 120 EC C₁₈色谱柱（100 mm × 4.6 mm，2.7 μm）分离，采用甲醇和5 mmol/L乙酸铵-0.1%甲酸溶液梯度洗脱，设置电雾式检测器的雾化温度为55℃，幂律参数（PFV）为1.35。甘氨胆酸、猪脱氧胆酸、胆酸、甘氨脱氧胆酸、鹅脱氧胆酸和脱氧胆酸分别在2.10~972 μg/mL、2.09~1046 μg/mL、2.05~978 μg/mL、2.56~1022 μg/mL、1.25~658 μg/mL和2.05~1025 μg/mL浓度范围内线性良好，相关系数（R²）大于0.9991，检出限在0.86~1.53 μg/mL之间，定量限在1.20~2.56 μg/mL之间，平均回收率在94.2%~105.3%之间，相对标准偏差（RSD）小于1.6%。将本方法应用于实际牛黄解毒片样品的测定，结果表明，不同厂家生产的牛黄解毒片中胆汁酸的含量差异显著。建立的6种胆汁酸的HPLC-CAD含量测定方法灵敏、准确、可靠，可用于中药复方牛黄解毒片质量评价。

关键词:

English

Determination of Six Kinds of Bile Acids in NiuhuangJiedu Tablets Using High Performance Liquid Chromatography Coupled with Charged Aerosol Detector

1.
Shanghai Key Laboratory of Functional Materials Chemistry, School of Chemistry and Molecular Engineering, East China University of Science and Technology, Shanghai 200237, China;
2.
Thermo Fisher Scientific China Co. Ltd., Shanghai 200000, China;
3.
Shanghai Key Laboratory of New Drug Design, School of Pharmacy, East China University of Science and Technology, Shanghai 200237, China

Corresponding author: HU Ping, huping@ecust.edu.cn

Received Date: 31 December 2021
Revised Date: 27 February 2022
Fund Project:
Supported by the National Natural Science Foundation of China (No.81973285).

Abstract: A high performance liquid chromatography-charge aerosol detector (HPLC-CAD) method for determination of six kinds of bile acids(Glycocholicacid, hyodeoxycholic acid, cholic acid, glycodeoxycholic acid, chenodeoxycholic acid and deoxycholic acid) in Niuhuang Jiedu tablets was developed. The tablet samples were extracted by ultrasonication in 70% methanol and concentrated under nitrogen flow. The separation of bile acids was achieved on an Agilent Poroshell 120 EC C₁₈ column (4.6 mm × 100 mm, 2.7 μm) with gradient elution using methanol and aqueous solution containing 0.1% formic acid and 5 mmol/L ammonium formate. CAD evaporation temperature was set at 55℃ and the power function value (PFV) was 1.35. The linearity ranges were 2.10-972 μg/mL for glycocholic acid, 2.09-1046 μg/mL for hyodeoxycholic acid, 2.05-978 μg/mL for cholic acid, 2.56-1022 μg/mL for glycodeoxycholic acid, 1.25-658 μg/mL for chenodeoxycholic acid and 2.05-1025 μg/mL for deoxycholic acid, respectively. All relative coefficients (R²) were higher than 0.9991. The limits of detection and the limits of quantitation ranged from 0.86 to 1.53 μg/mL and 1.20 to 2.56 μg/mL, respectively. The average recoveries of six kinds of bile acids were in the range of 94.2%-105.3% with RSD ≤ 1.63%. The method was used to determine the real tablet samples, and the results indicated that the contents of six bile acids had an obvious difference among the Niuhuang Jiedu tablets of different manufacturers. The HPLC-CAD method for the determination of six kinds of bile acids was sensitive, accurate and reliable, which could be applied to evaluate the quality of the Niuhuang Jiedu tablets.

Key words:

Niuhuang Jiedu tablet
/ High performance liquid chromatography
/ Charged aerosol detector
/ Bile acid

1. [1]
  Chinese Pharmacopoeia Comission. Chinese Pharmacopoeial (2020, Part I). Beijing:China Medical Science Press, 2020:694. 国家药典委员会. 中华人民共和国药典(2020年版, 一部). 北京:中国医药科技出版社, 2020:694.
2. [2]
  HU Z, HE L C, ZHANG J, LUO G A.J. Chromatogr. B:Anal. Technol. Biomed. Life Sci., 2006, 837(1-2):11-17.HU Z, HE L C, ZHANG J, LUO G A.J. Chromatogr. B:Anal. Technol. Biomed. Life Sci., 2006, 837(1-2):11-17.
3. [3]
  ZHANG Yu-Jing, XIA Jing, QIU Jia-Si, CAO Shuai, MAO Xiu-Hong, JI Shen. J. Int. Pharm. Res., 2016, 43(2):268-274. 张宇静, 夏晶, 仇佳思, 曹帅, 毛秀红, 季申. 国际药学研究杂志, 2016, 43(2):268-274.
4. [4]
  CHARLES T, ROBERTO P, MARK P, JOHAN A, KRISTINA S. Nat. Rev. Drug Discovery, 2008, 7(8):678-693.CHARLES T, ROBERTO P, MARK P, JOHAN A, KRISTINA S. Nat. Rev. Drug Discovery, 2008, 7(8):678-693.
5. [5]
  YANG F, HE Y Q, LIU H X, TSUEI J, JIANG X Y, YANG L, WANG Z T, WAN Y J Y. Biochem. Pharmacol., 2014, 91(4):483-489.YANG F, HE Y Q, LIU H X, TSUEI J, JIANG X Y, YANG L, WANG Z T, WAN Y J Y. Biochem. Pharmacol., 2014, 91(4):483-489.
6. [6]
  PERINO A, POLS T W H, NOMURA M, STEIN S, PELLICCIARI R, SCHOONJANS K. J. Clin. Invest., 2014, 124(12):5424-36.PERINO A, POLS T W H, NOMURA M, STEIN S, PELLICCIARI R, SCHOONJANS K. J. Clin. Invest., 2014, 124(12):5424-36.
7. [7]
  REITER S, DUNKEL A, DAWID C, HOFMANN T. J. Agric. Food Chem., 2021, 69(36):10572-10580.REITER S, DUNKEL A, DAWID C, HOFMANN T. J. Agric. Food Chem., 2021, 69(36):10572-10580.
8. [8]
  ASHLEY C J, OMAR G Q, VALERIE S F, PHILIPPE Z, CARMELO Q, LUIGI B, ANNE T, JULIE C, DANIELA F, MARCUS H, CATHERINE P, VINCENT S, CAMILLE A, SANDRINE Q, VALENTINE G, NATHALIE H, ALESSIA P, ALEXIA D, MARLENE M, THIERRY L L, SAMANTHA C, NATHALIE D, ASTRID C, DELPHINE G, BENOIT D, GILLES M, DAVID D, FREDRIK B, VINCENT P, GIOVANNI M, BART S, KRISTINA S, DANIELA C. Cell Metab., 2021, 33(7):1483-1492.ASHLEY C J, OMAR G Q, VALERIE S F, PHILIPPE Z, CARMELO Q, LUIGI B, ANNE T, JULIE C, DANIELA F, MARCUS H, CATHERINE P, VINCENT S, CAMILLE A, SANDRINE Q, VALENTINE G, NATHALIE H, ALESSIA P, ALEXIA D, MARLENE M, THIERRY L L, SAMANTHA C, NATHALIE D, ASTRID C, DELPHINE G, BENOIT D, GILLES M, DAVID D, FREDRIK B, VINCENT P, GIOVANNI M, BART S, KRISTINA S, DANIELA C. Cell Metab., 2021, 33(7):1483-1492.
9. [9]
  BETHAN L C, LESLIE R S, KEVIN J W, PAULINE M, ANGELA C, KELSEY E J, ALVIVN P C, MADELAINE C B S, ANNIKA W, JULIANNE W A, WILLIAM B, JAMES W, ANNA C C, LIU Y Y, ANDERS T, PETER J M, BRIAN G D, JULIA J M, HANNS U M, ELIZABETH J T, PETER A E, THOMAS Q A V. Cell Metab., 2021, 33(8):1671-1684.BETHAN L C, LESLIE R S, KEVIN J W, PAULINE M, ANGELA C, KELSEY E J, ALVIVN P C, MADELAINE C B S, ANNIKA W, JULIANNE W A, WILLIAM B, JAMES W, ANNA C C, LIU Y Y, ANDERS T, PETER J M, BRIAN G D, JULIA J M, HANNS U M, ELIZABETH J T, PETER A E, THOMAS Q A V. Cell Metab., 2021, 33(8):1671-1684.
10. [10]
  LUO M M, YAN J B, WU L Y, WU J T, CHEN Z, JIANG J P, CHEN Z Y, HE B H. J. Immunol. Res., 2021, 2021:2264737.LUO M M, YAN J B, WU L Y, WU J T, CHEN Z, JIANG J P, CHEN Z Y, HE B H. J. Immunol. Res., 2021, 2021:2264737.
11. [11]
  WU Y M, WANG X C, WU Q Z, WU X P, LIN X C, XIE Z H. Anal. Methods, 2010, 2(12):1927-1933.WU Y M, WANG X C, WU Q Z, WU X P, LIN X C, XIE Z H. Anal. Methods, 2010, 2(12):1927-1933.
12. [12]
  SHI Y, XIONG J, SUN D M, LIU W, WEI F, MA S C, LIN R C. J. Sep. Sci., 2015, 38(16):2753-2762.SHI Y, XIONG J, SUN D M, LIU W, WEI F, MA S C, LIN R C. J. Sep. Sci., 2015, 38(16):2753-2762.
13. [13]
  KONG W J, JIN C, LIU W, XIAO X H, ZHAO Y L, LI Z L, ZHANG P, LI X F. Food Chem., 2010, 120(4):1193-1200.KONG W J, JIN C, LIU W, XIAO X H, ZHAO Y L, LI Z L, ZHANG P, LI X F. Food Chem., 2010, 120(4):1193-1200.
14. [14]
  XIONG J, ZHENG T J, SHI Y, WEI F, MA S C, HE L, WANG S C, LIU X S. J. Pharm. Biomed. Anal., 2019, 174:50-56.XIONG J, ZHENG T J, SHI Y, WEI F, MA S C, HE L, WANG S C, LIU X S. J. Pharm. Biomed. Anal., 2019, 174:50-56.
15. [15]
  CHEN M J, LIU C, SHEN Y M, ZOU J F, ZHANG Z M, WAN Y, YANG L, JIANG S, QIAN D W, DUAN J N. J. Chromatogr. Sci., 2021, 59(9):871-876.CHEN M J, LIU C, SHEN Y M, ZOU J F, ZHANG Z M, WAN Y, YANG L, JIANG S, QIAN D W, DUAN J N. J. Chromatogr. Sci., 2021, 59(9):871-876.
16. [16]
  SHI Yan, WEI Feng, LV Bao-Jun, JIN Yan-Fang, CHEN Nan, DING Fang-Fang, MA Shuang-Cheng. Chin. Pharm. J., 2021, 56(10):790-793. 石岩, 魏锋, 吕宝俊, 靳艳仿, 陈楠, 丁芳芳, 马双成. 中国药学杂志, 2021, 56(10):790-793.
17. [17]
  KAKIYAMA G, MUTO A, TAKEI H, NITTONO H, MURAI T, KUROSA W A T, HOFMANNA F, PANDAK W M, BAJAJJS J. Lipid Res., 2014, 55(5):978-990.KAKIYAMA G, MUTO A, TAKEI H, NITTONO H, MURAI T, KUROSA W A T, HOFMANNA F, PANDAK W M, BAJAJJS J. Lipid Res., 2014, 55(5):978-990.
18. [18]
  ZHENG J P, YE C, HU B F, YANG H B, YAO Q F, MA J, LIU Y, LIU H T. Biotechnol. Appl. Biochem., 2020, 68(6):1332-1341.ZHENG J P, YE C, HU B F, YANG H B, YAO Q F, MA J, LIU Y, LIU H T. Biotechnol. Appl. Biochem., 2020, 68(6):1332-1341.
19. [19]
  LIU W X, CHENG X L, GUO X H, HU X R, WEI F, MA S C. Pharm. Biomed. Anal., 2020, 179:1-8.LIU W X, CHENG X L, GUO X H, HU X R, WEI F, MA S C. Pharm. Biomed. Anal., 2020, 179:1-8.
20. [20]
  SHAFAEI A, REES J, CHRISTOPHERSEN C T, DEVINE A, BROADHURST D, BOYCE M C. Anal. Chim. Acta, 2021, 1150:1-9.SHAFAEI A, REES J, CHRISTOPHERSEN C T, DEVINE A, BROADHURST D, BOYCE M C. Anal. Chim. Acta, 2021, 1150:1-9.
21. [21]
  YANG M T, TAN D P, LU A J, QIN L, WANG C H, LING H, LU Y L, HE Y Q. Int. J. Anal. Chem., 2021, 2021:5209618.YANG M T, TAN D P, LU A J, QIN L, WANG C H, LING H, LU Y L, HE Y Q. Int. J. Anal. Chem., 2021, 2021:5209618.
22. [22]
  WU L L, ZHANG S N, ZHOU L H, XIONG H S, GONG X C, ZHANG S J, PAN J Y, QU H B. Phytochem. Anal., 2021, 32(6):942-956.WU L L, ZHANG S N, ZHOU L H, XIONG H S, GONG X C, ZHANG S J, PAN J Y, QU H B. Phytochem. Anal., 2021, 32(6):942-956.
23. [23]
  LIU A X, XU T T, YANG W N, ZHOU D D, SHA Y W. J. Anal. Methods Chem., 2021, 2021:6616854.LIU A X, XU T T, YANG W N, ZHOU D D, SHA Y W. J. Anal. Methods Chem., 2021, 2021:6616854.
24. [24]
  XIE M J, YU Y T, ZHU Z Y, DENG L P, REN B, ZHANG M. J. Pharm. Biomed. Anal., 2021, 201:114087.XIE M J, YU Y T, ZHU Z Y, DENG L P, REN B, ZHANG M. J. Pharm. Biomed. Anal., 2021, 201:114087.
25. [25]
  XU X Y, WANG S M, WANG H M, HU W D, HAN L F, CHEN B X, LI X, WANG H D, LI H F, GAO X M, GUO D, YANG W Z. J. Chromatogr. A, 2021, 1655:462504.XU X Y, WANG S M, WANG H M, HU W D, HAN L F, CHEN B X, LI X, WANG H D, LI H F, GAO X M, GUO D, YANG W Z. J. Chromatogr. A, 2021, 1655:462504.
26. [26]
  XIE Q, TIAN H M, HUAN X H, CAO L L, WANG Y, CHENG X M, NING C G, HU F D, WANG C H. Phytochem. Anal., 2022, 33:262-271.XIE Q, TIAN H M, HUAN X H, CAO L L, WANG Y, CHENG X M, NING C G, HU F D, WANG C H. Phytochem. Anal., 2022, 33:262-271.
27. [27]
  NGUYEN H T, VU-HUYNH K L, NGUYEN H M, LE H T, LE T H V, PARK J L, NGUYEN M D. Molecules, 2021, 26(17):5373.NGUYEN H T, VU-HUYNH K L, NGUYEN H M, LE H T, LE T H V, PARK J L, NGUYEN M D. Molecules, 2021, 26(17):5373.
28. [28]
  WANG C, CHAO I C, QIN Y, ZHANG W X, ZHAO J, ZHANG Q W. J. Pharm. Biomed. Anal., 2022, 210:114545.WANG C, CHAO I C, QIN Y, ZHANG W X, ZHAO J, ZHANG Q W. J. Pharm. Biomed. Anal., 2022, 210:114545.
29. [29]
  VERTZONI M, ARCHONTAKI H, REPPAS C. J. Lipid Res., 2008, 49(12):2690-2695.VERTZONI M, ARCHONTAKI H, REPPAS C. J. Lipid Res., 2008, 49(12):2690-2695.
30. [30]
  WANG Yi-Bo, WANG Chun-Yu, QU Fan-Na, ZHANG Li-Ying, LIANG Li-Na, YU Yan-Hai, PAN Yuan-Yuan. Chin. J. Anal. Chem., 2014, 42(1):109-112. 王一博, 王春雨, 曲范娜, 张丽英, 梁丽娜, 余彦海, 潘媛媛. 分析化学, 2014, 42(1):109-112.
31. [31]
  PAWELLEK R, MUELLNER T, GAMACHE P, HOLZGRABE U. J.Chromatogr. A, 2021, 1637:461844.PAWELLEK R, MUELLNER T, GAMACHE P, HOLZGRABE U. J.Chromatogr. A, 2021, 1637:461844.
32. [32]
  GAMACHE P H. Charged Aerosol Detection for Liquid Chromatography and Related Separation Techniques. New Jersey:John Wiley & Sons, 2017.GAMACHE P H. Charged Aerosol Detection for Liquid Chromatography and Related Separation Techniques. New Jersey:John Wiley & Sons, 2017.
33. [33]
  AHMAD I A H, BLASKO A, WANG H, LU T, MANGION I, REGALADO E L. J. Chromatogr. A, 2021, 1641:461997.AHMAD I A H, BLASKO A, WANG H, LU T, MANGION I, REGALADO E L. J. Chromatogr. A, 2021, 1641:461997.
34. [34]
  Chinese Pharmacopoeia Comission. Chinese Pharmacopoeial (2020, Part IV). Beijing:China Medical Science Press, 2020:480. 国家药典委员会. 中华人民共和国药典(2020年版, 四部). 北京:中国医药科技出版社, 2020:480.

扫一扫看文章

计量

PDF下载量: 13
文章访问数: 687
HTML全文浏览量: 145

通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142

高效液相色谱-电雾式检测器测定牛黄解毒片中的6种胆汁酸

通讯作者: 胡坪,E-mail:huping@ecust.edu.cn

English

Determination of Six Kinds of Bile Acids in NiuhuangJiedu Tablets Using High Performance Liquid Chromatography Coupled with Charged Aerosol Detector

Corresponding author: HU Ping, huping@ecust.edu.cn

CCS Chemistry：嵌段共聚物自组装成 “三明治”二维有机材料，实现纳米级压力传感功能

CCS Chemistry：颜徐州、鲍哲南：你的皮肤可能是一片SPMs

CCS Chemistry：工作高效不疲劳，只须让催化剂动起来

CCS Chemistry：静电组装导向聚合- 聚电解质纳米凝胶量化制备新策略

CCS Chemistry：金黄色葡萄球菌醛基脱氢酶是如何识别特异性底物的？

计量

通讯作者: 陈斌, bchen63@163.com

中国化学会秘书处

高效液相色谱-电雾式检测器测定牛黄解毒片中的6种胆汁酸

通讯作者: 胡坪,E-mail:huping@ecust.edu.cn

English

Determination of Six Kinds of Bile Acids in NiuhuangJiedu Tablets Using High Performance Liquid Chromatography Coupled with Charged Aerosol Detector

Corresponding author: HU Ping, huping@ecust.edu.cn

CCS Chemistry：嵌段共聚物自组装成 “三明治”二维有机材料，实现纳米级压力传感功能

CCS Chemistry：颜徐州、鲍哲南： 你的皮肤可能是一片SPMs

CCS Chemistry：工作高效不疲劳，只须让催化剂动起来

CCS Chemistry：静电组装导向聚合- 聚电解质纳米凝胶量化制备新策略

CCS Chemistry：金黄色葡萄球菌醛基脱氢酶是如何识别特异性底物的？

计量

文章相关

通讯作者: 陈斌, bchen63@163.com

中国化学会秘书处

CCS Chemistry：颜徐州、鲍哲南：你的皮肤可能是一片SPMs