基于指纹图谱结合化学计量学及多成分含量测定的天麻蜜环菌粉质量控制研究

Study on the Quality Control of Gastrodia Tuder Halimasch Powder Based on Fingerprint Combined with Stoichiometry and Multi-component Content Determination

目的建立天麻蜜环菌粉HPLC指纹图谱,结合化学计量学筛选差异性成分,明确其化学成分的含量及潜在质量差异标志物。方法采用HPLC法,色谱柱为Symmetry Shield RP 18(250 mm×4.6 mm,5μm),流动相为乙腈-水溶液,梯度洗脱,体积流量为0.5 mL/min,检测波长为254 nm。对10批天麻蜜环菌粉的指纹图谱,采用《中药色谱指纹图谱相似度评价系统(2012版)》,指认共有峰并进行相似度评价;采用SPSS 26.0软件和SIMCA-P 14.1软件进行聚类分析(CA)、正交偏最小二乘判别分析(OPLS-DA);采用同一HPLC法测定8种成分的含量。结果建立了天麻蜜环菌粉指纹图谱,确定了13个共有峰,并指认出其中8个化学成分,分别为尿嘧啶(2号峰)、胞苷(3号峰)、次黄嘌呤(4号峰)、尿苷(5号峰)、黄嘌呤(6号峰)、5-羟甲基糠醛(10号峰)、鸟苷(11号峰)、腺苷(13号峰);10批样品相似度≥0.968;聚类分析及正交偏最小二乘判别分析结果一致,可将样品分为3组,筛选出7个主要差异性成分,其中6个为尿嘧啶、胞苷、次黄嘌呤、尿苷、鸟苷、腺苷;前期对指认出8个化学成分含量测定及方差分析的显著性与CA和OPLS-DA判别分析结果一致。结论研究建立的天麻蜜环菌粉HPLC指纹图谱结合化学计量学可阐明天麻蜜环菌粉质量特征及差异性,为天麻蜜环菌粉质量控制及探究天麻蜜环菌粉潜在质量标志物提供参考。

Objective To establish HPLC fingerprint of Gastrodia Tuder Halimasch Powder,combined with stoichiometry to screen the different components,and identify the content of chemical components and potential quality difference markers.Methods The chromatographic column was Symmetry Shield RP 18(250 mm×4.6 mm,5μm).The mobile phase was acetonitrile-water solution with gradient elution.The volume flow rate was 0.5 mL/min and the detection wavelength was 254 nm.For the fingerprints of 10 batches of Gastrodia Tuder Halimasch Powder,common peaks were identified and similarity evaluation was carried out using the Similarity Evaluation System of TCM Chromatographic Fingerprint(2012 Edition).SPSS 26.0 software and SIMCA-P 14.1 software were used for cluster analysis(CA)and orthogonal partial least squares discriminant analysis(OPLS-DA).The same HPLC method was used to determine the content of 8 components.Results The fingerprint of Gastrodia Tuder Halimasch Powder was established,13 common peaks were identified,and 8 chemical components were identified,namely uracil(peak 2),cytidine(peak 3),hypoxanthine(peak 4),uridine(peak 5),xanthine(peak 6),5-hydroxymethylfurfural(peak 10),guanosine(peak 11)and adenosine(peak 13).The similarity of 10 batches of samples≥0.968;The results of cluster analysis and orthogonal partial least squares discriminant analysis were consistent.The samples could be divided into 3 groups and 7 main different components were screened out,6 of which were uracil,cytidine,hypoxanthine,uridine,guanosine and adenosine.The significance of content determination and variance analysis of 8 chemical components was consistent with the results of CA and OPLS-DA discriminant analysis.Conclusion The established HPLC fingerprint combined with stoichiometry can clarify the quality characteristics and differences of Gastrodia Tuder Halimasch Powder,and provide reference for the quality control of Gastrodia Tuder Halimasch Powder and the exploration of potential quality markers of Gastrodia Tuder Halimasch Powder.