摘要
目的:基于高效液相色谱(HPLC)指纹图谱及聚类分析对市售阿胶进行质量评价,以期探索可靠的质量控制方法。方法:收集市售的10批阿胶商品药材,采用盐酸水解法制备水解氨基酸样品溶液,以异硫氰酸苯酯(PITC)和三乙胺柱前衍生化后进行HPLC分析。采用Welch ultimate amino acid(4.6 mm×250 mm,5μm)色谱柱,流动相A为0.1 mol/L醋酸钠溶液(调pH至6.5)-乙腈(93∶7)、流动相B为乙腈-水(80∶20),流速为1.0 ml/min,检测波长为254 nm,柱温为40℃。经梯度洗脱,得到各批阿胶中氨基酸含量,测定不同批次阿胶药材中17种氨基酸类成分的含量;使用相似度评价软件及SPSS 21.0软件进行相似度分析和聚类分析(HCA)。结果:阿胶中氨基酸衍生溶液36 h内保持稳定,17种氨基酸在线性范围内相关系数r均>0.995,加样回收率在85.8%~120.2%;10批阿胶的氨基酸指纹图谱具有专属性,相似度均>0.95,聚类分析结果与相似度结果相似。结论:该方法可靠,操作简便,精密度和重复性良好,可用于阿胶中17种水解氨基酸含量测定;10批阿胶中所含的氨基酸分布及比例比较稳定,将指纹图谱与聚类分析运用于不同来源阿胶的质量鉴别,可为药材特征评价方法的建立提供依据。
Objective： To evaluate the quality of commercially available Colla Corii Asini based on high performance liquid chromatography （HPLC） fingerprint and cluster analysis, and explore the reliable method for quality control. Methods： 10 batches of commercial medicinal material of CoUa Corii Asini were collected from the market. The sample solution of hydrolyzed amino acids was prepared by hydrochloric acid hydrolysis. Alter the pre column derivatization with phenyl isothioeyanate（ PITC ）and triethylamine, the HPLC analysis was performed on a Welch ultimate amino acid column （4.6 mm×250 mm,51μm）. The mobile phase A was 0.1 mol/L sodium acetate solution （ pH 6.5 ） -acetonitrile （93：7 ） and the mobile phase B was acetonitrile-water （ 80 ： 20）. The flow rate was 1.0 ml/min, the detection wavelength was 254 nm, and the column temperature was 40℃. The amino acid contents were detected after gradient elution, and the contents of 17 kinds of amino acid in different batches of medicinal material were determined. The similarity analysis and hierarchical clustering analysis（ HCA ） were conducted by similarity evaluation system for chromatographic fingerprint of TCM and SPSS 21.0 software. Results： The solution of amino acids derivatives in Colla Corii Asini remained stable within 36 hours. Within the linear range, the correlation coefficients of 17 kinds of amino acid were all more than 0. 995 ,and the recovery rate was 85.8% -120.2%. The fingerprint of amino acids in 10 batches of Colla Corii Asini was specific. All the similarities were greater than 0.95. The results of cluster analysis were similar to that of similarity analysis. Conclusion： The method is reliable, simple and convenient with good accuracy and repeatability ,which can be used in the content determination of 17 kinds of hydrolytic amino acids in Colla Corii Asini. The distribution and proportion of amino acids in 10 batches of Colla Corii Asini are relatively stable. The fingerprint and cluster analysis can be used f
出处
《上海中医药大学学报》
CAS
2017年第6期91-96,共6页
Academic Journal of Shanghai University of Traditional Chinese Medicine
基金
上海中医药大学第9批大学生科创项目(SHUTCM2016199)
