朱砂安神丸毛细管电泳指纹图谱研究

Capillary electrophoresis fingerprints of Zhusha Anshen Wan

目的建立朱砂安神丸(ZSASW)毛细管电泳指纹图谱(capillary electrophoresis fingerprint,CEFP),以系统指纹定量法鉴定其质量,并针对ZSASW-HPCE对照指纹图谱进行不确定度和可靠度评价。方法采用未涂层毛细管(75 cm×75μm,有效长度62 cm),以50 mmol.L-1硼砂-150 mmol.L-1磷酸二氢钠-50 mmol.L-1磷酸氢二钠(体积比为4∶1∶1,pH9.0)为背景电解质(BGE),运行电压12 kV,紫外检测波长203 nm,重力进样20 s。通过对12批样品聚类分析确定用其中9批生成对照CEFP(RCEFP),计算其不确定度和可靠度。以此RCEFP为标准鉴别12批样品质量。结果以小檗碱(BBR)为参照物峰,确定21个共有峰,用系统指纹定量法鉴别出S1、S2、S6、S10和S11质量很好,S5和S9质量好,S12质量良好,S3质量中等,S4、S7和S8质量劣。RCEFP的宏定性宏定量可靠度均>0.96。结论所建立的CEFP稳定可靠,可为ZSASW质量控制提供新参考。

Objective To establish the capillary electrophoresis fingerprint（CEFP） of Zhusha Anshen Wan（ZSASW）,identify the quality of ZSASW with systematic quantified fingerprint method and evaluate the uncertainty and reliability of ZSASW-HPCE referential fingerprint.Methods The electrophoretic separation was performed on a 75 cm ×75 μm column（with effective length of 62 cm） uncoated fused silica capillary with 50 mmol·L^-1 sodium borate-150 mmol·L^-1 sodium dihydrogen phosphate-50 mmol·L^-1 disodium hydrogen phosphate（4∶1∶1,v/v） adjusting to pH 9.0 as the background electrolyte.The running voltage was 12 kV,the wavelength was 203 nm at room temperature,and the time for gravity injection was 20 s.After performing cluster analysis on 12 batches of ZSASW,9 batches were selected to establish the referential CEFP（RCEFP）.The uncertainty and reliability of RCEFP were calculated.The ZSASW quality was evaluated by RCEFP as the criterion.Results Twenty-one co-possessing peaks were selected as the fingerprint peaks of ZSASW when taking berberine peak as the referential peak.Meanwhile,the 12 batches of ZSASW were assessed by systematic quantified fingerprint method,showing that S1,S2,S6,S10 and S11 were very good,S5 and S9 were good,S12 was fine,S3 was moderate,S4,S7 and S8 were inferior.The macro qualitative reliability and macro quantitative reliability of RCEFP were more than 0.96.Conclusion The established CEFP is reliable and can serve as a novel reference to identify and control the quality of ZSASWs.