水线草质量控制体系构建（英文）

Establishing a Quality Control System of Hedyotis corymbosa

目的水线草为中国等东南亚国家的主要抗肿瘤药物,但是目前没有切实可行的质量标准对其进行有效地控制,本研究欲构建水线草全面的质量控制体系。方法采用显微鉴别法对水线草进行粉末和横切面性状鉴别;采用薄层色谱法进行定性鉴别;采用高效液相色谱法建立水线草区别于易混淆品种白花蛇舌草的特征图谱,且对特征峰成分耳草酮A进行定量研究;采用2015年版中国药典规定的方法分别对水线草的水分、总灰分、酸不溶性灰分、重金属和农药残留量进行检测。结果水线草横切面内含草酸钙针晶束,偶见小油滴。薄层色谱法检出水线草样品中区别于白花蛇舌草的3个蓝色荧光斑点;高效液相色谱法(HPLC)建立了含5个特征峰的水线草特征图谱,并测定主要特征峰耳草酮A,该成分在0.0000527～0.00211 mg/mL范围内线性关系良好,线性方程为Y=3.15×10~8X+312.04(r^2=1.000),平均回收率为100.4%,RSD为0.85%,精密度(RSD 0.92%)、重复性(RSD 0.14%)、稳定性(RSD 0.85%)都达标,15批水线草中耳草酮A的质量分数在0.00017～0.00113%;水分、总灰分、酸不溶性灰分分别为8.80%～11.2%、7.00%～15.3%和0.440%～7.70%;重金属铅、砷、汞、镉、铜的含量分别为1.60～4.60、0.650～1.60、0.0300～0.100、0.240～0.690和10.1～16.6μg/g;农药残留含量低于检测限0.00100 ppm。结论此质量标准体系可有效区分水线草和易混淆品种白花蛇舌草,从而全面控制水线草质量,保证水线草的真伪优劣,为水线草临床使用提供参考。

Objective Hedyotis corymbosa(Shui Xian Cao,水线草,HC)is the main antitumor medicine in China and other southeast Asian countries.A comprehensive quality control system of HC was established in this study,as the current system was ineffective.Methods Microscopy was used to observe the characteristics of the powder and cross-sectional tissue.Thin layer chromatography(TLC)was employed to separate HC from Hedyotis diffusa(Bai Hua She She Cao,白花蛇舌草,HD).Simultaneously,high-performance liquid chromatography(HPLC)was used to establish the characteristic spectrum of HC and determine the concentrations of hedyotiscone A.Regarding the issue of systematicness,concentrations of water,total ash,acid-insoluble ash,heavy metals(including lead,arsenic,mercury,cadmium and copper)and pesticide residue were measured by the methods described in the 2015 edition of Chinese Pharmacopoeia.Results Calcium oxalate needle crystal bundles and oil drops were observed in the cross-sectional tissue.Three characteristic blue fluorescent spots were observed in the HC samples by TLC.There were five distinct characteristic peaks in HC but not in HD,and hedyotiscone A was the sharpest characteristic peak.The generated linear equation of hedyotiscone A was Y=3.15×108X+312.04(r2=1.000)between 0.00005270 and 0.002108 mg/mL,and the average recovery was 100.42%.Precision[relative standard deviation(RSD)0.92%],repeatability(RSD 0.14%),and recovery(RSD 0.85%)met the requirements for quantitative analysis.Hedyotiscone A concentrations ranged from 0.00017%~0.00113%across different habitats.Concentrations of water,ash and acid-insoluble ash were 8.80%~11.2%,7.00%~15.3%and 0.440%~7.70%,respectively.Concentrations of Pb,As,Hg,Cd and Cu were 1.60~4.60,0.650~1.60,0.0300~0.100,0.240~0.690 and 10.1~16.6μg/g,respectively.Pesticide residues were less than 0.00100 ppm.Conclusion The quality control system can effectively distinguish HC from HD,so as to comprehensively control the quality of HC,to provide references for the clinical use of HC.