[关键词]
[摘要]
目的 建立不同产地甘松Nardostachys jatamansi的HPLC指纹图谱及多成分含量测定方法,并结合化学模式识别法探究不同产地甘松化学成分差异,评价不同产地、不同批次甘松药材的质量。方法 采用HPLC法建立甘肃、青海、四川16批次甘松的指纹图谱,并对指标成分进行含量测定。进行相似度评价结合聚类分析(cluster analysis,HCA)、主成分分析(principal component analysis,PCA)和正交偏最小二乘法-判别分析(orthogonal partial least squares-discriminant analysis,OPLS-DA)等模式识别技术进行分析。结果 16批次甘松HPLC指纹图谱共匹配到60个共有峰,通过与对照品比对,指认出绿原酸、丁香酸、蒙花苷、甘松新酮、马兜铃酮5个色谱峰,指纹图谱相似度范围为0.943~0.999。16批次甘松样品中绿原酸、丁香酸、蒙花苷、甘松新酮、马兜铃酮含量测定变化范围分别为1.046 5~4.975 3、0.028 4~0.108 7、0.016 7~0.157 4、7.442 1~37.869 1、0.673 7~4.306 5 mg/g,5种成分在不同产地间均具有显著性差异。HCA将16批次甘松分为3类;PCA得不同批次甘松质量差异较大;经OPLS-DA筛选出了绿原酸、丁香酸、甘松新酮、马兜铃酮等31个差异成分。指纹图谱综合评价结果表明S4、S7、S10甘松样本质量较好。结论 确定了绿原酸、丁香酸、甘松新酮、马兜铃酮4种指标成分作为评价甘松质量差异的潜在标志性成分,建立的甘松HPLC指纹图谱及多成分含量测定方法稳定、可靠,可为甘松药材的质量控制、综合评价和临床应用的有效性提供科学依据。
[Key word]
[Abstract]
Objective To establish the HPLC fingerprints of Gansong (Nardostachys jatamansi DC.) from different origins and determine the content of multi-components, and to investigate the differences of the chemical components of N. jatamansi from different origins in combination with the chemical pattern recognition method, so as to evaluate the quality of N. Jatamansi from different origins and batches of N. jatamansi. Methods The fingerprints of 16 batches of N. jatamansi from Gansu, Qinghai and Sichuan were established by HPLC method, and the content determination of the index components was carried out. Similarity evaluation combined with pattern recognition techniques such as cluster analysis (HCA), principal component analysis (PCA) and orthogonal partial least squares-discriminant analysis (OPLS-DA) were performed. Results The HPLC fingerprints of 16 batches of N. jatamansi were matched to a total of 60 common peaks, and five peaks were identified as chlorogenic acid, cedar acid, acaciin, nardosinone and (−)-aristolone by comparison with the control product, and the similarity of fingerprints was in the range of 0.943—0.999. The variations of the contents of chlorogenic acid, cedar acid, acaciin, nardosinone and (−)-aristolone in 16 batches of N. jatamansi samples were in the range of 1.046 5—4.975 3 mg/g, 0.028 4—0.108 7 mg/g, 0.016 7—0.157 4 mg/g, 7.442 1—37.869 1 mg/g, and 0.673 7—4.306 5 mg/g respectively, and the five components were significantly different among different origins. HCA divided the 16 batches of N. jatamansi into three categories; PCA showed that the quality of different batches of N. jatamansi varied greatly; 31 differential components, including chlorogenic acid, cedar acid, nardosinone and (−)-aristolone, were screened out by OPLS-DA. The results of the comprehensive evaluation of fingerprint mapping showed that the quality of S4, S7 and S10 samples of N. jatamansi was better. Conclusion Four index components, chlorogenic acid, cedar acid, nardosinone and (−)-aristolone, were identified as the potential marker components for evaluating the quality difference of N. jatamansi, and the established HPLC fingerprints of N. jatamansi and the method for determining the content of multi-components were stable and reliable, which can provide scientific basis for the quality control, comprehensive evaluation and the effectiveness of the clinical application of N. jatamansi medicinal herbs.
[中图分类号]
[基金项目]
2022年度甘肃省中医药科研立项课题项目(GZKP-2022-39);甘肃中医药大学科学研究与创新基金重点项目(2020KCZD-2);甘肃省科技计划项目(东西协作专项-鲁甘科技协作)(22CX8NA068);甘肃省教育厅双一流重大科研项目(GSSYLXM-05)