[高效液相色谱法对藏药手参中4种活性成分的定量分析]

[Quantitative analysis of four active constituents in Tibetan herb Gymnadenia conopsea by high-performance liquid chromatography].

作者信息

Yang Beibei, Li Shuai, Zhang Ruiping, Wang Yan, Shi Jiangong

机构信息

Institute of Materia Medica, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100050, China.

出版信息

Zhongguo Zhong Yao Za Zhi. 2009 Jul;34(14):1819-22.

PMID:19894516

Abstract

OBJECTIVE

To develop a high performance liquid chromatography method for simultaneous determination of tour active constituents, dactylorhin B, loroglossin, dactylorhin A, militarine present in a Tibetan herb Gymnadenia conopsea.

METHOD

The analysiswas achieved on a Pravil C18 column (4.6 mm x 150 mm, 5 microm) eluted with the mobile phases of methanol (A)-water (B) in gradient elution. The initial condition was 15% A, chsnhrf to 20% A in 10 min, to 45% A in 10 min, to 70% A in 15 min. The detection wavelength was set at 222 nm. The flow rate was 0.7 mL x min(-1). Orthogonal test was adopted to optimize extraction process of four active compounds from Gymnadenia conopsea.

RESULT

The assay displayed good linearity over the concentration ranges of 0.20-4.05 microg (r = 0.9999), 0.10-5.11 micro g (r = 0.9999), 0.10-5.11 microg (r = 0.9999) and 0.09-1.79 microg (r = 0.9999), respectively. The average recoveries (n=9) were 99.19%, 99.16%, 99.52% and 99.00% for dactylorhin B, loroglossin, dactylorhin A and militarine respectively.

CONCLUSION

The method is simple, sensitive, reliable and reproducible which can be used for the quality study of Gymnadenia conopsea.

摘要

目的

建立一种高效液相色谱法，用于同时测定藏药手参中4种活性成分，即细距手参苷B、舌唇兰苷、细距手参苷A和手参碱。

方法

采用Pravil C18柱（4.6 mm×150 mm，5μm），以甲醇（A）-水（B）为流动相进行梯度洗脱。初始条件为15% A，10分钟内升至20% A，再10分钟内升至45% A，15分钟内升至70% A。检测波长设定为222 nm。流速为0.7 mL·min⁻¹。采用正交试验优化手参中4种活性成分的提取工艺。

结果

该测定方法在0.20 - 4.05μg（r = 0.9999）、0.10 - 5.11μg（r = 0.9999）、0.10 - 5.11μg（r = 0.9999）和0.09 - 1.79μg（r = 0.9999）浓度范围内分别呈现良好的线性关系。细距手参苷B、舌唇兰苷、细距手参苷A和手参碱的平均回收率（n = 9）分别为99.19%、99.16%、99.52%和99.00%。