七清败毒颗粒质量标准研究

Study on the quality standard of Qiqing Baidu granules

为完善七清败毒颗粒的质量标准,为其质量评价提供依据,采用反相高效液相色谱法建立七清败毒颗粒特征图谱,标识方中黄芩、虎杖、板蓝根、大青叶各种组分群体特征。通过查阅相关文献及预分析试验,确定以黄芩(黄芩苷)、虎杖(虎杖苷与大黄素)、板蓝根(靛玉红)、大青叶(靛玉红)为指标测定成分。采用Waters XBridge C_(18)色谱柱(4.6 mm×250.0 mm,3.5μm),以乙腈为流动相A,以0.4%磷酸溶液为流动相B,进行梯度洗脱,采用PDA检测器,3D扫描190~450 nm范围,2D检测波长为288 nm,流速为1.0 mL/min,柱温为30℃。供试品色谱中呈现4个特征峰,并与对应的对照品色谱峰保留时间一致,经过精密度试验、稳定性试验、耐用性试验证明该方法灵敏度高,专属性强,结果重复性好。将混合对照品溶液及6批七清败毒颗粒样品的色谱图数据导入“中药色谱指纹图谱相似度评价系统(2012版)”,以S1为参照图谱,采用平均数相关系数法进行多点校正,时间窗为0.1 s,进行自动匹配,计算得6批七清败毒颗粒与对照图谱之间的相似度均大于0.993;通过色谱图与光谱图比对,对照品溶液与七清败毒颗粒样品中相应的4个共有峰均有良好的重合性,符合《中华人民共和国兽药典》要求。该方法节约了试剂、时间,操作简便、环保,可以更有效地对七清败毒颗粒内在质量进行控制。

In order to improve the quality standard of Qiqing Baidu granules and provide a basis for its quality evaluation,the characteristic chromatogram of Qiqing Baidu granules was established by RP-HPLC to identify the population characteristics of various components of Radix Scutellariae,Rhizoma Polygoni Cuspidati,Radix Isatidis and Folium Isatidis.The components of baicalin,polydatin and indatin were determined by referring to relevant literatures and pre-analysis tests.The chromatographic column was Waters XBridge C_(18)(4.6 mm×250.0 mm,3.5μm),and acetonitrile was used as mobile phase A.Gradient elution was performed with 0.4%phosphoric acid solution as mobile phase B.PDA detector was used to scan the range of 190-450 nm in 3D,and the detection wavelength was determined to be 288 nm,the flow rate was 1.0 mL/min,and the column temperature was 30℃.The chromatogram showed 4 characteristic peaks,and the retention time was consistent with the corresponding reference peaks.The precision test,stability test and durability test proved that the method had high sensitivity,strong specificity and good repeatability.The chromatographic data of mixed reference solution and 6 batches of Qiqing Baidu granules samples were imported into the TCM Chromatographic Fingerprint Similarity Evaluation System(2012 edition).With S1 as the reference graph,the mean correlation coefficient method was used for multi-point correction,and the time window was 0.1 s for automatic matching.The similarity between 6 batches of Qiqing Baidu granules and the control graph was calculated.All of them were greater than 0.993.The corresponding 4 common peaks in the reference solution and Qiqing Baidu granules sample were compared by chromatogram and spectrogram,and all of them had good coincidence,which met the requirements of Chinese Veterinary Pharmacopoeia.The method saves reagents and time,is easy to operate and environmentally friendly,and can control the internal quality of Qiqing Baidu granules more effectively.