基于~1H-NMR代谢组学技术比较熏硫和未熏硫白芷的化学成分差异及化学标志物筛选

Comparison of Chemical Constituents Differences between Angelicae dahuricae with and without Sulphurfumigation and Selection of Chemical Markers Based on^1H-NMR Metabolomics

目的:研究熏硫与未熏硫白芷药材的整体化学成分差异,并筛选可用于两者鉴别的化学标志物。方法:分别以硫熏和未硫熏的白芷药材各16批为检测样品,以氘代甲醇-重水(1∶0.2,V/V)为提取溶剂,采用氢核磁共振(1H-NMR)代谢组学技术检测两种方法处理的白芷药材中的初级和次级代谢产物。利用直观分析、主成分分析(PCA)和t检验比较二者的化学成分差异,并确定化学标志物。结果:从两种白芷药材中同时检测并鉴定出了包括香豆素类、氨基酸类、糖类在内的19种化学成分。直观分析发现,两种白芷药材的化学成分轮廓相似,但香豆素类和糖类成分的1H-NMR信号强度有较大的差异;PCA分析结果显示,所有样品按不同处理方法分成两类;t检验结果显示,白芷熏硫后氧化前胡素、欧前胡素、葡萄糖和蔗糖含量均显著降低(P<0.01或P<0.001),丙氨酸、亮氨酸含量均显著升高(P<0.01或P<0.001),而其余13种成分含量差异无统计学意义(P>0.05)。结论:熏硫和未熏硫白芷药材中氧化前胡素、欧前胡素、葡萄糖和蔗糖、丙氨酸、亮氨酸6种化学成分存在明显的差异,可作为化学标志物用于市场上熏硫白芷的鉴别。

OBJECTIVE:To study the differences in the overall chemical constituents of Angelicae dahuricae with and without sulphur-fumigation,and to select chemical markers that can be used for identification.METHODS:Each 16 batches of A.dahuricae with and without sulphur-fumigation were selected as samples.Deuterated methanol-deuterium oxide(1∶0.2,V/V)was used as extraction solvent,and^1H-NMR metabolomics was used to detect the primary and secondary metabolites in A.dahuricae.In addition,visual analysis,principal component analysis(PCA)and t-test were used to compare the component differences of A.dahuricae by two kinds of pretreatment methods.RESULTS:A total of 19 chemical constituents such as coumarins,amino acids and sugars were simultaneously detected and identified from two kinds of A.dahuricae.Visual analysis showed that the chemical profiles of the two kinds of A.dahuricae were similar,but their coumarins and carbohydrates were quite different in 1H-NMR signal intensity.PCA analysis showed that all samples could be divided into two categories according to different treatment methods.The results of t-test showed that the contents of oxypeucedanin,imperatorin,glucose and sucrose of A.dahuricae were decreased significantly after sulphur fumigation(P<0.01 or P<0.001),while the contents of alanine and leucine were significantly increased(P<0.01 or P<0.001).There was no statistical significance in the contents of other 13 kinds of components(P>0.05).CONCLUSIONS:There are obvious differences in the 6 chemical compounds of oxypeucedanin,imperatorin,glucose,sucrose,alanine and leucine in A.dahuricae with and without sulphur-fumigation,which can be used as chemical markers for the identification of sulphur-fumigated A.dahuricae in the market.