目的:比较发酵对天麻Halimasch粉(GTHP)化学成分的影响,建立其指纹图谱和多组分含量测定,并为处理提供基础,处理,以及该草药的临床应用。
方法:采用超高效液相色谱-四极杆-Orbitrap高分辨质谱(UHPLC-Q-OrbitrapHRMS)对发酵前后GTHP中的化学成分进行了初步分析。采用高效液相色谱法(HPLC)测定GTHP的主要成分并建立指纹图谱。聚类分析(CA),采用主成分分析(PCA)进行综合评价。
结果:确定了79种化合物,包括类黄酮,有机酸,核苷,萜类化合物,和其他人。CA和PCA结果显示10个样本分为3组。通过标准对照和HPLC分析,从22个峰中鉴定出10个化合物,即尿嘧啶,鸟苷,腺苷,5-羟甲基糠醛(5-HMF),Daidzin,Genistin,糖素,Daidzein,Genistein,和麦角甾醇.发酵后,GTHP表现出明显较高的尿嘧啶含量,鸟苷,腺苷,5-羟甲基糠醛,和麦角甾醇,并显着降低染料木素和大豆黄酮的含量。
结论:UHPLC-Q-OrbitrapHRMS和HPLC方法可有效鉴定GTHP发酵前后的多种化学成分。本研究为进一步研究GTHP的合理临床应用和质量控制改进提供了有价值的参考。
OBJECTIVE: To compare the effect of fermentation on the chemical constituents of Gastrodia Tuder Halimasch Powder (GTHP), to establish its fingerprinting and multicomponent content determination, and to provide a basis for the processing, handling, and clinical application of this herb.
METHODS: Ultra-high-performance liquid chromatography-quadrupole-Orbitrap high-resolution mass spectrometry (UHPLC-Q-Orbitrap HRMS) was used to conduct a preliminary analysis of the chemical constituents in GTHP before and after fermentation. High-performance liquid chromatography (HPLC) was used to determine some major differential components of GTHP and establish fingerprints. Cluster analysis (CA), and principal component analysis (PCA) were employed for comprehensive evaluation.
RESULTS: Seventy-nine compounds were identified, including flavonoids, organic acids, nucleosides, terpenoids, and others. The CA and PCA results showed that ten samples were divided into three groups. Through standard control and HPLC analysis, 10 compounds were identified from 22 peaks, namely
uracil, guanosine, adenosine, 5-hydroxymethylfurfural (5-HMF), daidzin, genistin, glycitein, daidzein, genistein, and ergosterol. After fermentation, GTHP exhibited significantly higher contents of
uracil, guanosine, adenosine, 5-hydroxymethylfurfural, and ergosterol and significantly lower genistein and daidzein contents.
CONCLUSIONS: The UHPLC-Q-Orbitrap HRMS and HPLC methods can effectively identify a variety of chemical components before and after the fermentation of GTHP. This study provides a valuable reference for further research on the rational clinical application and quality control improvement of GTHP.