不同年份老香黄定量分析及其化学模式识别研究

建立了一种联合化学标志物定量分析和化学模式识别技术快速判别不同年份老香黄的方法,并筛选其差异性化学标志物。采用超高效液相色谱-串联质谱(UPLC-MS/MS)建立了对21种与老香黄相关的化学标志物及其内标物山柰酚-3-O-芸香糖苷的检测方法,该方法的线性关系良好,灵敏度高,专属性强,可用于对不同年份老香黄的区别分析。根据检出的不同年份老香黄的15个化学标志物含量,结合主成分分析(PCA)、聚类分析(HCA)、正交偏最小二乘法判别分析(OPLS-DA)法进行分析,可快速判别不同年份的老香黄,并筛选出6个差异性化学标志物。结果表明,贡献最大的差异化学标志物为5,7-二甲氧基香豆素,其中0年含量为(47.465±1.987)μg/g,3年含量为(17.069±0.542)μg/g,5年含量为(11.073±1.862)μg/g,8年含量为(41.905±2.240)μg/g,10年含量为(21.079±2.270)μg/g,15年含量为(43.267±2.432)μg/g,20年含量为(51.711±1.439)μg/g,该标志物的含量在一定程度上体现了不同年份的老香黄,可用于区分老香黄的贮藏年份。为了验证方法的可靠性,进行盲样适用性能力测试,测定结果所得年份和厂家提供的保持一致。该研究给市面上老香黄的质量控制及其品质鉴定提供了快速可靠的方法。

Quantitative Analysis and Chemical Pattern Recognition of Lao-Xiang-Huang Preserved in Different Years

A quantitative analysis method of chemical markers coupled with pattern recognition algorithms was established for the rapid discrimination of Lao-Xiang-Huang preserved in different years.A ultra performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS)method was developed for the analysis of 21 chemical markers related to Lao-Xiang-Huang and the internal standard kaempferol-3-O-rutinoside.The developed method was applied to the distinguishing of different preserving years of Lao-Xiang-Huang with good linearity,high sensitivity and strong specificity.According to the amounts of 15 detected chemical markers and combining analysis of principal component analysis(PCA),hierarchical cluster analysis(HCA)and orthogonal partial least squares discriminant analysis(OPLS-DA),six differential chemical markers were identified significantly in contributing to the discrimination.Results showed that the chemical marker 5,7-dimethoxycoumarin was the uppermost contributor in distinguishing the preserving year of Lao-Xiang-Huang,which amounts were(47.465±1.987)μg/g in the 0th year,(17.069±0.542)μg/g in the 3rd year,(11.073±1.862)μg/g in the 5th year,(41.905±2.240)μg/g in the 8th year,(21.079±2.270)μg/g in the 10th year,(43.267±2.432)μg/g in the 15th year and(51.711±1.439)μg/g in the 20th year.The amount of 5,7-dimethoxycoumarin could reflect the preserving year of Lao-Xiang-Huang in a certain extent.To verify the reliability of the developed method,the proficiency test was carried out,and the calibrated year was in line with that provided by the manufacturer.This study provides a rapid and reliable method for the quality identification and control of Lao-Xiang-Huang in the market.