泽泻炮制前后化学成分及其利尿作用研究

目的分析比较泽泻饮片麸炒、盐炙、土炒、酒炙炮制前后14种三萜类成分的含量,探讨泽泻及其不同炮制品的利尿作用。方法采用超高液相色谱-质谱联用(UPLC-MS),色谱柱为Waters CORTECS UPLC C18(2.1 mm×100 mm,1.6μm),以乙腈-水为流动相进行梯度洗脱,单四级杆质谱采用正离子SIR模式,测定泽泻生品和炮制品饮片中16-羰基-23-乙酰泽泻醇A、16-羰基-24-乙酰泽泻醇A、泽泻醇C、泽泻醇F、23-乙酰泽泻醇C、泽泻醇L、24-乙酰泽泻醇F、泽泻醇A、23-乙酰泽泻醇A、24-乙酰泽泻醇A、泽泻醇G、泽泻醇B、23-乙酰泽泻醇B、11-去氧泽泻醇B共14种三萜类成分的含量,探讨泽泻炮制过程中化学成分含量变化;采用水负荷大鼠代谢笼法,比较泽泻炮制前后的利尿作用。结果经方法学验证,所建立UPLC-MS同时测定泽泻饮片中14种三萜类成分的方法符合分析要求。与生品比较,泽泻炮制品化学成分最主要的变化是泽泻三萜类24、25位环氧环发生开环转化,23-乙酰泽泻醇B、泽泻醇B、11-去氧泽泻醇B和23-乙酰泽泻醇C含量下降,而泽泻醇A类化合物含量升高,且盐炙泽泻的化学成分转化最为明显。动物实验结果显示:生品组和各炮制品组1 h尿量较空白对照组显著增加,且利尿效应酒炙品>土炒品>麸炒品>盐炙品>生品;生品组和盐炙品组2 h尿量与空白对照组比较差异无统计学意义,而酒炙品组、土炒品组、麸炒品组尿量较空白对照组显著增加,且利尿效应酒炙品>土炒品>麸炒品。结论本研究建立的UPLC-MS测定泽泻生品及不同炮制品中化学成分含量方法准确、快速。泽泻炮制后化学成分发生明显变化,泽泻经酒炙、土炒和麸炒后较生品利尿作用显著增强。

Study on Changes of Chemical Composition and Diuretic Effect in Alismatis Rhizoma after Processing

Objective To analyze the contents of 14 kinds of triterpenoids composition and diuretic effect of Alismatis Rhizoma before and after processing,including stir-frying with bran,stir-frying with salt-water,stir-frying with soil,and stir-frying with wine.Methods The UPLC-MS method was performed on a Waters CORTECS UPLC C18 column(2.1 mm×100 mm,1.6μm)with a gradient elution using acetonitrile and water to quantitative analysis of the 14 triterpenoids(16-oxo-23-acetate-alisol A,16-oxo-24-acetate-alisol A,alisol C,alisol F,alisol C 23-acetate,alisol L,alisol F 24-acetate,alisol A,alisol A 23-acetate,alisol A 24-acetate,alisol G,alisol B,alisol B 23-acetate,11-deoxy-alisol B)of crude Alismatis Rhizoma and processed products.The method of metabolizable cage of rats was adopted to study the difference of diuretic effect of Alismatis Rhizoma before and after processing.Results Through methodological verification,the method of simultaneous determination of 14 triterpenoids in Alismatis Rhizoma by the established UPLC-MS met the analytical requirements.Compared with crude Alismatis Rhizoma,the most important processing change in the 14 triterpenoids of Alismatis Rhizoma was the ring-opening at 24,25 epoxy rings in the triterpenoids.The contents of alisol B 23-acetate,alisol B,11-deoxy alisol B and alisol C 23-acetate decreased,and the content of alisol A increased.The conversion was most significant in stir-frying with salt-water.The results of the rats experiment showed that compared with the control group,the urine output of crude and processed products of Alismatis Rhizoma significantly increased within 1 h,and the diuretic effect of Alismatis Rhizoma processed products,stir-frying with wine>stir-frying with soil>stir-frying with bran>stir-frying with salt-water>crude Alismatis Rhizoma;there was no statistical significance in urine output of crude and stir-frying with salt-water within 2 h,Alismatis Rhizoma processed products of stir-frying with wine,soil,and bran significantly increased compared with the control group,and the diuretic effect of stir-frying with wine>stir-frying with soil>stir-frying with bran.Conclusion The established UPLC-MS method is accurate and rapid for the determination the chemical composition of different processed products of Alismatis Rhizoma.The chemical composition of Alismatis Rhizoma processed products changes after the processing,and the diuretic effect of the stir-frying with wine,soil and bran of Alismatis Rhizoma increases.