基于HPLC的益脑心颗粒特征图谱分析方法的建立

Establishment of HPLC characteristic chromatograms of Yinaoxin granule

目的建立益脑心颗粒的高效液相色谱(HPLC)特征图谱分析方法。方法采用HPLC法,色谱条件:Kromasil C_(18)柱(250 mm×4.6 mm,5μm);流动相为乙腈-0.1%磷酸水溶液,梯度洗脱;流速为1.0 ml/min;检测波长为270 nm;柱温为30℃。结果依据中药色谱特征图谱相似度评价系统(2012版),以二苯乙烯苷为参照峰,通过方法学研究精密度、稳定性与重复性结果良好,确定了益脑心颗粒的HPLC特征图谱共有模式,标定了35个特征峰。10批制剂整体相似度在0.900-0.998之间。对35个特征峰进行指认,鉴别出8个特征峰即腺苷、天麻素、没食子酸、5-羟甲基糠醛、巴利森苷A、二苯乙烯苷、迷迭香酸、丹酚酸B。结论建立了益脑心颗粒HPLC特征图谱分析方法,该方法简便可靠、重复性好,为本品的质量控制提供了科学方法,为制剂临床研究提供依据。

Objective To establish a method for analyzing the characteristic chromatogram of Yinaoxin granules by HPLC.Methods HPLC was performed on Kromasil C_(18)column(250 mm×4.6 mm,5μm).The acetonitrile-0.1%phosphoric acid aqueous solution was used as mobile phase for gradient elution.The flow rate was 1.0 ml/min,and the detection wavelength was 270 nm,and the column temperature was 30℃.Results According to the Similarity Evaluation System of Traditional Chinese Medicine Chromatogram(2012 Edition),the methodological study was carried out with stilbene glycoside as the reference peak,and the results were good in precision,stability and repeatability.The common mode of HPLC chromatogram of Yinaoxin granules was determined,and 35 characteristic peaks were calibrated.The overall similarity of 10 batches of preparations was 0.900-0.998.The sources of 35 characteristic peaks were identified,and eight characteristic peaks were identified,namely adenosine,gastrodin,gallic acid,5-hydroxymethylfurfural,palisenoside A,stilbene glycoside,rosmarinic acid and salvianolic acid B.Conclusion The established method is simple,reliable and reproducible,which provides a scientific method for the quality control of Yinaoxin granules and a basis for clinical research.