HPLC法同时测定粉防己中汉防己甲素和汉防己乙素的含量

宋俊蓉 娄华勇 方友来 蓝俊杰 何敏 马小攀 潘卫东

摘 要：基于更好的完善粉防己的質量控制标准，以提取时间、方法、溶剂比为主要影响因素，考察了不同提取方法对粉防己中汉防己甲、乙素提取率的影响，建立了一种同时测定粉防己中汉防己甲素和汉防己乙素的高效液相色谱分析方法。色谱条件为：Hypersil GOLD Phenyi柱（250 mm × 4.6 mm， 5 μm），甲醇- 0.1% 三乙胺 （85∶15），检测波长为228 nm，进样量10μL，柱温30℃，流速1 mL/min。样品中汉防己甲素在0.12～2.31 μg范围内呈良好的线性关系，平均加样回收率分别为102.80%、104.46%、103.49%，RSD分别为1.72%、0.38%、1.39%；汉防己乙素在0.05～1.09 μg范围内呈良好的线性关系，平均加样回收率分别为101.00%、100.08%、100.00%，RSD分别为0.40%、1.82%、1.83%。结果表明江西的粉防己中汉防己甲素和汉防己乙素含量高于其他产地。上述测定方法操作便捷、重现性好、结果可靠。

关键词：高效液相色谱法；粉防己；汉防己甲素；汉防己乙素；同时测定

中图分类号：O656.22

文献标识码：A

文章编号：1008-0457（2018）03-0079-05 国际DOI编码：10.15958/j.cnki.sdnyswxb.2018.03.014

Simultaneous Determination of Tetrandrine and Fangchinoline from Stephania tetrandra by HPLC

SONG Junrong1，2， LOU Huayong2， FANG Youlai1，2， LAN Junjie1，2， HE Min2， MA Xiaopan3， PAN Weidong2*

（1. School of Pharmaceutical Sciences， Guizhou University， Guiyang， Guizhou 550002， China； 2. State Key Laboratory of Functions and Applications of Medicinal Plants， Guizhou Medical University， Guiyang， Guizhou 550014， China； 3. Zunyi Medical University， Zunyi， Guizhou 563000， China）

Abstract： In order to Improve the quality control standard of Stephania tetrandra， establish a high performance liquid chromatography （HPLC） method for simultaneously determination the content of Tetrandrine and Fangchinoline in S. tetrandra，was established. The effects of different extraction methods， the extraction time， method， and solvent ratio in particular， on the extraction rate of Tetrandrine and Fangchinoline were investigated. The conditions of HPLC were as below： Hypersil GOLD Phenyi column （250 mm ×4.6 mm， 5μm）， and a mobile phase consisted of methanol and 0.1% triethylamine （85∶15） by gradient elution at a flow rate of 1 mL/min. The detection wavelength was 228 nm and sample volume was 10 μL. The sample showed a good linear relationship with tetrandrine in the range of 0.12～2.31 μg. The mean sample recovery was 102.80%， 104.46%， and 103.49%， respectively， and the RSD was 1.72%， 0.38%， and 1.39%， respectively. The sample showed a linear relationship with Fangchinoline in the range of 0.05～1.09 μg. The average sample recovery rate was 101.00%， 99.08%， and 100.00%， respectively. The RSD was 0.40%， 1.8% and 0.40%， respectively. The above HPLC conditions were used to determine the contents of tetrandrine and Fangchinoline in S. tetrandra of different origins. The results showed that the contents of tetrandrine and Fangchinoline in S. tetrandra from Jiangxi Province were higher than those from other origins. The method described in the present study appears to be convenient， reproducible and reliable.

Key words：HPLC； Stephania tetrandra； tetrandrine； fangchinoline； simultaneous determination

粉防己（Stephania tetrandra S.Moore），又名汉防己、土防己、石蟾蜍、猪大肠等，为防己科植物千金藤属植物粉防己的干燥块根[1]，味苦辛，性寒，归膀胱、肺经，具有解热镇痛、利水消肿，祛风止痛等功效[2]，用于水肿脚气、小便不利、风湿痹痛、湿疹疮毒、高血压等症的治疗。粉防己中生物碱含量占总含量的1.5%～2.3% [3]，以双苄基异喹啉类生物碱为主 [4]，主要为汉防己甲素（Tetrandrine， 简称Tet）和汉防己乙素（Fangchinoline，简称Fan）。现代药理学证明粉防己中的生物碱具有良好的生物活性，在抗肝纤维化、拮抗Ca2+治疗高血压、诱导肿瘤细胞凋亡等方面都有较好的效果[5-6]，汉防己乙素亦可诱导肝癌细胞的自噬性死亡[7]。……