目的 通过OPA-FMOC在线自动衍生化,建立土鳖虫的HPLC指纹图谱分析方法。方法 色谱柱为Durashell-AA柱(150 mm×4.6 mm,3 μm);流动相组成为A:0.01 mol/L磷酸氢二钠及四硼酸钠水溶液(盐酸调节pH值至8.2);流动相B:甲醇-乙腈-水(45:45:10);体积流量1.6 mL/min;检测波长:0~24 min,338 nm;24~27 min,228 nm;柱温45℃;梯度洗脱。结果 共得到28个共有峰,指认了其中15个化学成分,建立了土鳖虫及其混淆品HPLC指纹图谱并计算相似度,根据SPSS聚类分析结果,将其分为2类,第Ⅰ类药材为地鳖或冀地鳖,第Ⅱ类为金边地鳖。结论 所建立的土鳖虫HPLC指纹图谱快速、准确、重复性好,为土鳖虫的质量控制提供科学依据。

Objective To establish the chromatographic fingerprint analysis for the quality control of Eupolyphaga steleophaga with OPA-FMOC by on-line derivatization. Methods Analysis was completed with a gradient elution of 0.01 mol/L sodium dihydrogen phosphate and 0.01 mol/L sodium tetraborate, pH=8.2(A) and methanol-acetonitrile-water was 45:45:10(B). The wavelength was set at 338 nm from 0-24 min, and at 228 nm from 24-27 min. Efficient and symmetrical peaks were obtained with data flowrate of 1.6 mL·min. Results A total of 28 peaks were detected, while 15 ingredients were identified. The 18 batches of samples were classified into two groups based on the results of cluster analysis. The first category was E. sinensis and Steleophaga plancyi; The second category was Opisthoplata orientalis and it was identified as the adulterants. Conclusion The HPLC fingerprint method is repeatable, feasible in analysis of E. steleophaga and could be used scientifically to evaluate the quality of E. steleophaga with quality control.

教育部科学技术研究重点项目(212032);辽宁省教育厅创新团队-中药质量控制关键技术(LT2012018)