Simultaneous determination of four active components in Alisma orientale (Sam.) Juz. by HPLC–DAD using a single reference standard

A rapid, simple and practical high-performance liquid chromatography method coupled with diode array detector（HPLC–DAD） was developed to evaluate the quality of Alisma orientale（Sam.） Juz.through a simultaneous determination of four major active triterpenes using a single standard to determine the multi-components（SSDMCs）. Alisol B 23-acetate was selected as the reference compound for calculating the relative response factors. All calibration curves showed good linearity（R240.9998） within test ranges. RSDs for intra- and inter-day of four analytes were less than 3.6% and 2.3%; the overall recovery was 92.1–110.2%（SSDMC）. The proposed method was successfully applied to quantify the four components in 20 samples from different localities in China. Moreover, significant variations were demonstrated in the content of these compounds. In addition, hierarchical clustering analysis（HCA） and principal components analysis（PCA） were performed to differentiate and classify the samples based on the contents of Alisol C 23-acetate, Alisol A, Alisol A 24-acetate and Alisol B 23-acetate. This simple,rapid, low-cost and reliable HPLC–DAD method using SSDMC is suitable for routine quantitative analysis and quality control of A. orientale（Sam.） Juz.

A UFLC/MS/MS method for simultaneous quantitation of alisol A and alisol B 23-acetate from Alisma orientale (Sam.) Juz. in rat plasma

A sensitive and reliable ultra fast liquid chromatography tandem mass spectrometry(UFLC-MS/MS)method has been developed and validated for simultaneous quantitation of alisol A and alisol B 23-acetate from Alisma orientale(Sam.)Juz.in rat plasma using diazepam as an internal standard(IS).The plasma samples were extracted by liquideliquid extraction with methyl tert-butyl ether and separated on a Venusil MP C18 column(100 mm×2.1 mm,3.0 mm)(Venusil,China)using gradient elution with the mobile phase consisting of methanol and 0.1%acetic acid in water at a flow rate of 0.4 ml/min.The two analytes were monitored with positive electrospray ionization by multiple reaction monitoring mode(MRM).The lower limit of quantitation was 5.00 ng/ml for alisol A and 5.00 ng/ml for alisol B 23-acetate.The calibration curves were linear in the range of 5.00 e2500 ng/ml for alisol A and 5e2500 ng/ml for alisol B 23-acetate.The mean extraction recoveries were above 63.8%for alisol A and 68.0%for alisol B 23-acetate from biological matrixes.Both intra-day and inter-day precision and accuracy of analytes were well within acceptance criteria(15%).The validated method was successfully applied to the pharmacokinetic study of alisol A and alisol B 23-acetate in rat plasma after oral administration of alcohol extract of Alismatis Rhizoma.

Amelioration effects of traditional Chinese medicine on alcohol-induced fatty liver

AIM： To examine the effects of traditional Chinese medicine （TCM） on alcohol-induced fatty liver in rats. TCM consists of Astragalus membranaceus, Morus alba, Crataegus pinnatifida, Alisma orientale, Salvia miltiorrhiza, and Pueraria lobata. METHODS： The rats were separated randomly into five groups. One （the CD group） was fed a control diet for 10 wk, another （the ED group） fed an ethanol-containing isocaloric liquid diet for 10 wk, and the last three （the TCM group） were fed an ethanol-containing isocaloric liquid diet for 10 wk and dosed orally with TCM （222 mg/kg.d, TCM222; 667 mg/kg·d, TCM667; and 2 000 mg/kg.d, TCM2000, respectively） weekly during the last 4 wk. RESULTS： ED group developed fatty liver according to lipid profile and liver histological findings. Compared with the control group, liver/body weight, serum triglyceride （TG） and total cholesterol （TC）, liver TG and TC, serum alanine aminotransferase （ALT） and aspartic aminotransferase （AST） significantly increased in the ED group. Whereas, in the rats administered with TCM, liver/body weight, serum TG and TC, liver TG and TC, serum ALT and AST were significantly decreased, and the degree of hepatic lipid droplets was markedly improved compared with those in the ED group. CONCLUSION： TCM treatment causes significant reduction in alcohol-induced lipid hepatic accumulation, reversing fatty liver and liver damage, and can be used as a remedy for alcoholic fatty liver.