三七伤药片超高效液相色谱特征图谱的研究

目的: 建立三七伤药片的UPLC特征图谱,为三七伤药片的质量评价提供参考。方法: 采用UPLC法建立特征图谱,通过液质联用对特征峰进行指认。采用ACQUITY UPLC BEH C₁₈(50 mm×2.1 mm,1.7 μm)色谱柱,以乙腈(A)-0.1%甲酸溶液(B)为流动相进行梯度洗脱(0~10 min,4%A→12%A;10~22 min,12%A→16%A;22~35 min,16%A→25%A),流速0.2 mL·min^-1,检测波长230 nm,柱温40 ℃;并采用电喷雾离子源进行正/负离子模式同时扫描,对特征峰进行指认。采用Chempattern^TM软件将三七伤药片的特征图谱生成共有模式,并进行PCA统计分析,以各主要色谱峰的保留时间和峰面积为变量得到Score图和Loading图。结果: 以柚皮苷峰为参照物峰建立三七伤药片UPLC特征图谱,确定了8个共有特征峰,并指认了新北美圣草苷、芍药苷、羟基红花黄色素A和柚皮苷4个特征峰。采用主成分分析(PCA)对特征图谱数据进行分析,结果表明不同样品化学成分存在差异,其中柚皮苷、新北美圣草苷和芍药苷的差异较大。结论: 建立的UPLC特征图谱方法具有较好的稳定性和重现性,能为三七伤药片的质量控制提供参考。

Objective: To establish a UPLC chromatographic fingerprint analysis method of Sanqishangyao tablets. Methods: The characteristic fingerprint was performed by HPLC and characteristic peaks were identified by LC-MS.The samples were separated on the ACQUITY UPLC BEH-C₁₈ column(50 mm×2.1 mm,1.7 μm),and eluted with a gradient elution(0-10 min,4%A→12%A;10-22 min,12%A→16%A;22-35 min,16%A→25%A)of acetonitrile as the mobile phase A and the solution containing 0.1% formic acid as the mobile phase B at a flow rate of 0.2 mL·min^-1.The detection wavelength was 230 nm and the column temperature was 40 ℃.According to the chempattern^TM advanced chemometrics solution,a common pattern was built and statistically analyzed by principal components analysis(PCA).The Score plot and th e Loading plot were obtained with the retention time and the peak area for each main chromatographic peak as the variable.Four characteristic peaks in the fingerprints were identified with applying electrospray ionization(ESI)source in the positive/negative ion mode. Results: The characteristic fingerprint of Sanqishangyao tablets was established with naringin as the reference peak.Four characteristic peaks were identified as hydroxysafflor yellow A,neoeriocitrin,naringin,paeoniflorin.Eight co-possessing peaks were designated as characteristic common peaks.The PCA showed the chemical compositions in the 27 samples were obviously different.The main difference of the chemical compositions was the content of neoeriocitrin,naringin and paeoniflorin. Conclusion: The fingerprint method can reflect the actual quality with good stability and reproducibility and be applied in the quality control of Sanqishangyao tablets.