诃子制草乌HPLC特征图谱研究及指标性成分定量测定

目的建立诃子制草乌的HPLC特征图谱及指标性成分的定量分析方法,分析辅料炮制对草乌物质基础的影响。方法采用HPLC色谱法,色谱柱Waters SunFire C_(18)(250 mm×4.6 mm,5μm);流动相为0.1%甲酸乙腈-10 mmol/L的乙酸铵溶液(含0.1%甲酸),梯度洗脱,体积流量1mL/min;柱温为35℃,进样量10μL。特征图谱检测波长为254 nm,定量检测波长分别为271 nm和235 nm,对诃子制草乌进行特征图谱分析及指标性成分的定量研究。结果建立了诃子制草乌的HPLC特征图谱,确定了28个共有峰,其中17个峰来自生草乌,11个峰来自炮制辅料诃子。对照品指认了新增加的6号峰和18号峰分别为没食子酸和诃子鞣酸。本试验建立的HPLC特征图谱分析方法可同时定量检测诃子制草乌中没食子酸、诃子鞣酸、苯甲酰新乌头原碱和新乌头碱的含量。结论建立的草乌及其炮制品的HPLC特征图谱方法简便、可靠,可对生草乌、制草乌及诃子制草乌进行快速鉴别区分,较为全面的反映草乌及不同炮制品的物质组成差异。

Specific analysis of Aconiti Kusnezoffii Radix processed with Fructus Chebulae with HPLC and quantification of its quality control ingredients

Objective To develop a high-performance liquid chromatography(HPLC), and conduct an qualitative analysis of quality control ingredients of Radix Aconiti Kusnezoffii (Kusnezoff Monkshood Root, Caowu) processed with Fructus Chebulae (Medicine Terminalia Fruit, Hezi), and analyzed the effects of auxiliary materials and processing on the material basis of Aconiti Kusnezoffii Radix (Zhicaowu).Methods HPLC specific chromatogram was performed using Waters SunFire C18 column (250 mm×4.6 mm, 5 μm), gradient elution using formic acid in acetonitrile (A) and 10 mmol/L ammonium acetate solution (B) with flow rate at 1 mL/min, column temperature at 35℃ and injection volume of 10 μL;detection wave length with a UV detector at 254 nm for fingerprint analysis, 271 nm and 235 nm for the determination.Results HPLC specific diagram of caowu processed with hezi was established, and 28 communal peaks were confirmed including 17 from caowu and 11 from hezi.Two new peaks (peak 6 and peak 16) were identified as gallic acid and chebulagic acid based on reference.This HPLC specific analysis method also quantified gallic acid, chebulagic acid, benzoylmesaconine and mesaconitine.Conclusion This specific HPLC method is easy to perform and also reliable.It could quickly identify raw, processed, and hezi-processed caowu, and distinguish different ingredients of caowu and its processed products.