^(1)H NMR ERETIC2结合多变量分析法定量分析生麻黄与蜜炙麻黄差异

Quantitative analysis of the difference between raw and honey-fried Ephedra sinica Stapf by ^(1)H NMR ERETIC2 combined with multivariate analysis

目的利用^(1)H NMR ERETIC2定量方法对14个批次的生麻黄和6个批次的蜜炙麻黄进行定性和定量分析,并筛选出差异成分,为麻黄质量标准的提升提供科学依据。方法以特征性成分麻黄碱信号强度为优选指标,对提取方法和复溶溶剂进行优化;建立基于^(1)H NMR ERETIC2方法的生麻黄和蜜炙麻黄特征成分绝对定量方法;采用^(1)H NMR谱图结合多变量分析法寻找不同批次生麻黄和蜜炙麻黄潜在的差异成分。结果最佳样品前处理方法为以甲醇为溶剂,物料比为1∶5(g/mL),超声提取5 min,氘代甲醇作为复溶溶剂。^(1)H NMR图谱共鉴定了11个化学成分,其中8个共有成分被选作定量指标。定量结果显示,生麻黄和蜜炙麻黄中异亮氨酸、蔗糖、葡萄糖和丙氨酸的含量差异有统计学意义(P<0.05);多元统计分析结果也表明,生麻黄和蜜炙麻黄之间存在明显差异,葡萄糖和蔗糖是主要的差异贡献成分,且在蜜炙麻黄中含量较高,而蜜炙麻黄中丙氨酸和异亮氨酸的含量则明显下降(P<0.05)。结论本研究建立的^(1)H NMR ERETIC2定量方法结合多元统计分析,可筛选出生麻黄和蜜炙麻黄的差异成分,能够有效区分生麻黄和蜜炙麻黄,为后续麻黄质量标准的完善提供了科学依据。

Objective To qualitatively and quantitatively analyze 14 batches of raw Ephedra sinica(E.sinica)Stapf and 6 batches of honey-fried E.sinica with ^(1)H NMR ERETIC2,and screen the differential components to provide a scientific basis for the improvement of quality standards of E.sinica.Methods The extraction method and the redissolve solvent were optimized with the signal intensity of the ephedrine as the index.A strategy for the absolute quantification of the characteristic components of raw and honey-fried E.sinica based on the ^(1)H NMR ERETIC2 method was established.^(1)H NMR spectroscopy together with multivariate analysis was utilized to determine the potential differential components in different batches of raw and honey-fried E.sinica.Results The best sample preparation method was optimized as follows:methanol as the solvent,the material-solvent ratio of 1∶5(g/mL),ultrasonic extraction for 5 min,and deuterated methanol as the redissolve solvent.Totally 11 components were identified by ^(1)H NMR spectroscopy,8 of which were common components were selected for quantification indexes.The quantitative analysis showed significant difference(P<0.05)in the content of isoleucine,sucrose,glucose and alanine between the raw and honey-fried E.sinica.The multivariate analysis indicated significant difference between the raw and honey-fried E.sinica.Glucose and sucrose were both higher in honey-fried E.sinica,while the content of alanine and isoleucine was significantly decreased(P<0.05).Conclusion The quantitative method of ^(1)H NMR ERETIC2 combined with multivariate statistical analysis helps reveal the differential components of raw and honey-fried E.sinica,which can effectively distinguish the raw and honey-fried E.sinica and provide a scientific basis for the subsequent improvement of quality control of E.sinica.