藏药菥蓂子的定性、定量方法研究

目的:建立藏药菥蓂子的定性与定量控制方法。方法:采用薄层色谱、高效液相色谱分别鉴定及测定15批菥蓂子中的黄酮类成分异牡荆素、当药黄素和硫代葡萄糖苷类成分黑芥子苷。两类成分薄层色谱鉴别的固定相分别为聚酰胺薄膜、高效硅胶GF254,展开剂分别为三氯甲烷-甲醇-冰醋酸(11∶1∶1,V/V/V)、乙酸乙酯-甲醇-三乙胺(4∶5∶0.5,V/V/V)。在异牡荆素、当药黄素含量测定的色谱条件中,色谱柱为CAPCELL PAK MGⅡC18,流动相为乙腈-0.4%冰醋酸溶液,梯度洗脱,流速为1.0 mL/min,检测波长为336 nm;在黑芥子苷含量测定的色谱条件中,色谱柱为CAPCELL PAK MGII C18,流动相为乙腈-0.02 mol/L四丁基硫酸氢铵水溶液(15∶85,V/V,pH 6),流速为1.0 mL/min,检测波长为227 nm。结果:在薄层色谱鉴别图谱中,供试品分别检测出与异牡荆素、当药黄素及黑芥子苷对照品相应的斑点;异牡荆素、当药黄素、黑芥子苷检测质量浓度线性范围分别为1.26~79.00、1.21~75.38、12.80~640.00μg/mL(r均≥0.999 5),检测限分别为0.09、0.12、0.15μg/mL,定量限分别为0.39、0.43、0.54μg/mL;精密度、稳定性(24 h)、重复性试验的RSD均≤2.0%(n=6),加样回收率分别为99.1%、97.0%、98.1%,RSD分别为1.9%、1.8%、1.8%(n=6)。15批菥蓂子药材中异牡荆素、当药黄素、黑芥子苷含量范围分别为0.013~0.090、0.020~0.130、18.92~40.75 mg/g。结论:建立的质量控制方法操作简便,重复性及稳定性良好,可用于藏药菥蓂子的质量控制。

Qualitative and Quantitative Study of Tibetan Medicine Thlaspi semen

OBJECTIVE:To establish the qualitative and quantitative control method of Tibetan medicine Thlaspi semen.METHODS:TLC and HPLC method were used to identify and determine flavonoids isovitexin,swertisin and glucosinolates sinigrin from 15 batches of T.semen.The stationary phases identified by TLC of flavonoids and glucosinolates were polyamide film and high performance silica gel GF254.The developing agents were trichloromethane-methanol-glacial acetic acid(11∶1∶1,V/V/V)and ethyl acetate-methanol-triethylamine(4∶5∶0.5,V/V/V).In chromatogram condition of content determination of isovitexin and swertisin,the separation was performed on CAPCELL PAK MGⅡC18 column with mobile phase composed of acetonitrile-0.4%glacial acetic acid solution(gradient elution)at the flow rate of 1.0 mL/min.The detection wavelength was set at 336 nm.In chromatogram condition of content determination of sinigrin,the separation was performed on CAPCELL PAK MGⅡC18 column with mobile phase composed of acetonitrile-0.02 mol/L tetrabutylammonium hydrogen sulfate(15∶85,V/V,pH 6)at the flow rate of 1.0 mL/min.The detection wavelength was set at 227 nm.RESULTS:In TLC identification chromatogram,spots corresponding to isovitexin,swertisin and sinigrin control were detected in test samples.The linear ranges of isovitexin,swertisin and sinigrin were 1.26-79.00,1.21-75.38,12.80-640.00μg/mL,respectively(all r≥0.999 5).The limits of detection(LODs)were 0.09,0.12,0.15μg/mL,and limits of quantitation(LOQs)were 0.39,0.43,0.54μg/mL,respectively.RSDs of precision,stability(24 h)and reproducibility tests were all lower than 2.0%(n=6).The recoveries were 99.1%,97.0%and 98.1%,and RSDs were 1.9%,1.8%,1.8%(n=6),respectively.The contents of isovitexin,swertisin and sinigrin in 15 batches of T.semen were 0.013-0.090,0.020-0.130 and 18.92-40.75 mg/g,respectively.CONCLUSIONS:Established quality control method is simple,reproducible and stable,and can be used for the quality control of Tibetan medicine T.semen.