胆南星发酵制品与混合蒸制品的鉴别研究

Study on Identification of Fermentation Products and Mixed Steamed Products of Arisaema Cum Bile

目的:以猪胆汁及天南星、制天南星为原料分别制备发酵制胆南星及混合蒸制胆南星,以胆汁酸类成分为指标,建立以猪胆汁为炮制辅料的胆南星2种炮制品的薄层鉴别及HPLC-ELSD特征图谱。方法:采用硅胶G薄层板,以石油醚:丙酮:冰醋酸(11∶8∶0. 8)为展开剂,10%硫酸乙醇溶液为显色剂,建立薄层色谱鉴别方法。采用Agilent Eclipse XDB C_(18)(4. 6 mm×250 mm,5μm)色谱柱,以乙腈与0. 1%冰醋酸水为流动相进行梯度洗脱,流速为1 m L/min,柱温为30℃,ELSD:漂移管温度为105℃,氮气流量为2. 5 m L/min,建立胆南星HPLC-ELSD特征图谱鉴别方法。结果:TLC和HPLC-ELSD图谱均显示发酵制胆南星可检测到游离型胆汁酸类成分猪胆酸(HCA)、猪去氧胆酸(HDCA)和鹅去氧胆酸(CDCA)3个特征成分,而混合蒸制胆南星中未检测出;混合蒸制胆南星可检测到结合型胆汁酸类成分甘氨猪去氧胆酸(GHDCA)和甘氨鹅去氧胆酸(GCDCA) 2个特征成分,而发酵制胆南星中未检测出。结论:所建胆汁酸类成分的薄层色谱鉴别法(TLC)和高效液相色谱-蒸发光散射法(HPLC-ELSD)均可有效鉴别胆南星的发酵制品和混合蒸制品,为胆南星质量控制提供依据。

Objective:Using pig bile and Arisaematis Rhizome and Arisaematis Rhizoma Preparatum as raw materials,the fermenting and mixed evaporating of arisaema cum bile were prepared respectively,and the bile acids were used as the index.To establish thin layer identification and HPLC-ELSD characteristic map of 2 processed products from porcine bile.Methods Silica gel G thin layer plate was applied,and petroleum ether:acetone:glacial acetic acid(11∶8∶0.8)were used as developing agent and 10%sulfuric acid ethanol solution as chromogenic agent,a TLC identification method was established.An Agilent Eclipse XDB C 18(4.6 mm×250 mm,5μm)column was used,with acetonitrile and 0.1%glacial acetic acid water as mobile phase for gradient elution.The flow rate was 1 mL/min,at 30℃and ELSD:drift tube at 105℃.The identification method of HPLC-ELSD characteristic map of arisaema cum bile was established by nitrogen flow rate of 2.5 mL/min.Results:TLC and HPLC-ELSD spectra showed that three characteristic components of free hyocholic acid(HCA),hyodeoxycholic acid(HDCA)and chenodeoxycholic acid(CDCA),could be detected in fermented arisaema cum bile,but not in mixed evaporation.Two characteristic components of conjugated bile acids,(GHDCA)and(GCDCA),could be detected by mixed evaporation of arisaema cum bile,but not in the fermentation of arisaema cum bile.Conclusion:TLC and HPLC-ELSD can be used to identify the fermented and mixed distilled products of arisaema cum bile which can provide the basis for the quality control of arisaema cum bile.