秦艽生药中龙胆苦苷和落干酸的含量及其HPLC指纹图谱测定

秦艽生药中龙胆苦苷和落干酸的含量及其HPLC指纹图谱测定

马晓庆, 邓伟峰, 杨东辉^*, 刘广学, 洪浩, 白光男, 蔡少青

1. 北京大学医学部药学院天然药物学系, 北京 100191
2. 怀化医学高等专科学校, 湖南怀化 418000
3. 平壤医科大学药学系, 朝鲜

收稿日期:2010-03-22 修回日期:2010-06-10 出版日期:2010-07-15 发布日期:2010-07-15
通讯作者: 杨东辉*

HPLC fingerprinting and quantification of gentiopicroside and loganic acid in Gentianae Macrophyllae Radix crude drugs

Xiao-Qing Ma, Wei-Feng Deng, Dong-Hui Yang^*, Guang-Xue Liu, Hao Hong,
Kuang-Nam Paek, Shao-Qing Cai

1. Department of Natural Medicines, School of Pharmaceutical Sciences, Peking University Health Science Center, Beijing 100191, China
2. Huaihua Medical Courses in General, Huaihua 418000, China
3. Pharmacy Department, Pyongyang Medical University, P.R.Korea

Received:2010-03-22 Revised:2010-06-10 Online:2010-07-15 Published:2010-07-15
Contact: Dong-Hui Yang*

摘要/Abstract

摘要： 本研究建立了秦艽药材中龙胆苦苷 (GPS) 和落干酸 (LA) 的含量测定及其HPLC指纹图谱分析方法。该方法采用Zorbax SB-C18 柱 (250 mm×4.6 mm, 5 μm), 0.04% 磷酸水 (A)-乙腈 (B) 为流动相梯度洗脱, 检测波长为230 nm, 流速为1.0 mL/min。利用L9 (3⁴) 正交试验确定药材的最佳提取工艺为: 每份秦艽样品 (g) 中加入15倍量的50% 乙醇 (mL), 超声提取30分钟。定量分析结果表明秦艽生药中龙胆苦苷的含量 (14.05 mg/g-74.61 mg/g) 明显高于落干酸 (1.13 mg/g-40.46 mg/g) 的含量。根据样品中龙胆苦苷和落干酸含量的比值 (1.8-11.4) 可将大叶秦艽 (GPS-LA 的比值≤4.3) 与细梗秦艽 (含小秦艽, GPS-LA的比值≥4.8) 进行区分。秦艽生药HPLC指纹图谱的主成分聚类分析结果也显示大叶秦艽与细梗秦艽 (含小秦艽) 可被分为两个不同的类群。本文所建立的HPLC-DAD 分析方法对于秦艽生药的品种鉴定和质量控制具有指导意义和参考价值。

关键词:

秦艽, HPLC指纹图谱, 龙胆苦苷, 落干酸, 主成分分析

Abstract:

HPLC fingerprinting and quantification of gentiopicroside (GPS) and loganic acid (LA) in Gentianae Macrophyllae Radix (GMR) crude drugs were developed in this study. The samples were separated on Zorbax SB-C18 column (250 mm×4.6 mm, 5 μm) with a linear gradient of acetonitrile and 0.04% phosphoric acid. The HPLC flow rate was 1.0 mL/min and a UV absorption was measured at 230 nm. An orthogonal L9 (3⁴) test was applied for the optimization of sample extraction conditions, and an aliquot of GMR sample (g) was extracted with 15-fold of 50% ethanol (mL) for 30 min by sonication. Quantitative analysis showed that the content of GPS (14.05 mg/g-74.61 mg/g) in all samples was obviously higher than that of LA (1.13 mg/g-40.46 mg/g). Based on the content ratio of GPS over LA (1.8-11.4), samples originated from Gentiana macrophylla (with content ratio of GPS over LA ≤4.3) could be distinguished from those from G. dahurica and G. dahurica var. gracilipes (with content ratio of GPS over LA ≥4.8). The principle components analysis of the HPLC fingerprints showed that samples originated from G. macrophylla and G. dahurica (including G. dahurica var. gracilipes) could be divided into two groups. This established HPLC-DAD method could be efficiently used for the species identification and quality control of GMR crude drugs.

Key words: Gentianae Macrophyllae Radix, HPLC fingerprint, Gentiopicroside, Loganic acid, Principle components analysis

中图分类号:

R282

Supporting:

Foundation item: Research Program of Science Technology of Traditional Chinese Medicine (TCM) Sponsored by the State Administration of TCM of China (Grant No. 04-05ZL01).
^*Corresponding author. Tel.: 86-10-82802534

马晓庆, 邓伟峰, 杨东辉^*, 刘广学, 洪浩, 白光男, 蔡少青

. 秦艽生药中龙胆苦苷和落干酸的含量及其HPLC指纹图谱测定[J]. .

Xiao-Qing Ma, Wei-Feng Deng, Dong-Hui Yang^*, Guang-Xue Liu, Hao Hong,
Kuang-Nam Paek, Shao-Qing Cai . HPLC fingerprinting and quantification of gentiopicroside and loganic acid in Gentianae Macrophyllae Radix crude drugs [J]. .

参考文献

[1] Jiangsu College of New Medicine. A Comprehensive Dictionary of the Traditional Chinese Medicines. Shanghai: Shanghai Science and Technology Press. 1977, 1764-1767.
[2] The State Pharmacopoeia Commission of China. Pharmacopoeia of the Peoples Republic of China. Beijing: Chemical Industry Press. 2005, 190-191.
[3] Dong, J.Y.; Li, G.Y.; Li, C.T. J. Gansu Coll. Tradit. Chin. Med. 2006, 23, 17-19.
[4] Kim, K.S.; Rhee, H.I.; Park, E.K.; Jung, K.; Jeon, H.J.; Kim, J.H.; Yoo, H.; Han, C.K.; Cho, Y.B.; Ryu, C.J.; Yang, H.I.; Yoo, M.C. Chin. Med. 2008, 3, 10.
[5] Yu, F.R.; Yu, F.H.; Li, R.D.; Wang, R. J. Ethnopharmacol. 2004, 95, 77-81.
[6] Liu, Y.H.; Li, X.C.; Liu, Y.Q.; Yang, C.R. Acta Bot. Yunnan. 1994, 16, 85-89.
[7] Sun, J.; Chen, G.C.; Li, Y.L.; Zhao, X.E.; Wang, H.L. Chin. J. Anal. Lab. 2006, 25, 28-31.
[8] Sun, J.; Li, Y.L.; Ji, K.J.; Ma, Y.H.; Xu, W.H.; Chen, G.C. Acta Bot. Yunnan. 2006, 28, 219-222.
[9] Tan, R.X.; Wolfender, J.L.; Zhang, L.X.; Ma, W.G.; Fuzzati, N.; Marston, A.; Hostettmann, K. Phytochemistry. 1996, 42, 1305-1313.
[10] Wu, Y.X.; Chen, G.; Yu, C.Y. J. Beijing Univ. Chem. Technol. (Nat. Sci. Ed.). 2008, 35, 64-67.
[11] Lin, P.C.; Ye, Z.H.; Lu, Y.C.; Zhao, S.Q.; Hu, F.Z.; Ji, L.J. Acta Bot. Yunnan. 2004, 27, 819-821.
[12] Chen, C.X.; Liu, Z.W.; Sun, Z.R.; Song, C.Q.; Hu, Z.B. Chin. Tradit. Herb. Drugs. 2003, 34, 814-816.
[13] Christopoulou, C.; Graikou, K.; Chinou, I. Chem. Biodivers. 2008, 5, 318-323.
[14] Kondo, Y.; TakaNo, F.; Hojo, H. Planta Med. 1994, 60, 414-416.
[15] Raj, K.; Vijayavitthal, T.M.; Bhaduri, A.P.; Pandey, C.P.; Patnaik, G.K. Indian J. Chem. 1996, 35(B), 1056-1061.
[16] Recio, M.C.; Giner, R.M.; Manez, S.; Rois, J.L. Planta Med. 1994, 60, 232-234.
[17] Zhang, M.J.; Wu, Q.X.; An, Y. Chin. Tradit. Herb. Drugs. 2007, 38, 759-761.
[18] Chen, Q.L.; Shi, Z.Y.; Sun, W.J. J. Northwest Univ. (Nat. Sci. Ed.). 2007, 37, 253-256.
[19] Wu, Q.X.; An, Y.; Zhang, M.J.; Lu, Y.C.; Bao, J.Y.; Hu, S.Q.; Yu, J. Acta Bot. Boreali. Occident. Sin. 2006, 35, 64-67.
[20] Lou, Z.C.; Qin, B. Species Systematization and Quality Evaluation of Commonly Used Chinese Traditional Drugs (North-China Edition, Volume 3), Beijing: Peking Medical College Peking Union Medical College United Press. 1996, 385-388.
[21] Nakamoto, K.; Otsuka, H.; Yamasaki, K. Phytochemistry. 1988, 27, 1856-1858.