Effect of postharvest processing on quality traits of Radix Gentianae Macrophyllae: A integrative analysis of metabolomics and proteomics.

The dried roots and rhizomes of Radix Gentianae Macrophyllae are widely used as food material or medicinal crops. "Sweating" is a traditional postharvest processing method, the basic processing procedure consists of softening, stacking and drying. The aim of this paper is to unveil the scientific connotation responsible for the "Sweating" processing in Radix Gentianae Macrophyllae during postharvest. Thus, the effect of different postharvest processing methods on the metabolic pathways of Radix Gentiasnae Macrophyllae was studied by the non-targeted metabolomic technique in combination with the label-free proteomics approach. The results showed that the differentially accumulated metabolites and abundant proteins were mainly enriched in the pathways of phenylalanine, tyrosine and tryptophan biosynthesis, polyphenols and terpenoids biosynthesis. "Sweating" has a greater up-regulation effect on these pathways than "Non-sweating", and can induce protein expression and metabolite accumulation associated with the quality traits of Radix Gentianae Macrophyllae. The results provide a detailed explanation of the scientific connotation of crucial steps of "Sweating" processing wherein opportunities existed for taking appropriate measures to enhance the accumulation of bioactive ingredients. These findings will serve as significant references for enhancing the postharvest processing technology of Radix Gentianae Macrophyllae and similar plants, resulting in higher product quality for food or plant materials production.

A new and effective evaluation method for Radix Gentianae Macrophyllae herbs based on 2-phenylethyl ß-d-glucopyranoside, 2-methoxyanofinic acid and gentioxepine by UPLC-QTOF-MS exploring and HPLC-DAD quantification.

Radix Gentianae Macrophyllae (RGM) is a traditional Chinese medicine belonging to the Gentiana genus and including four species of herbs. Owing to the lack of characteristic constituents, it is difficult to discriminate RGMs of different origins or even differentiate between herbs in the same genus. The current research aimed to explore the characteristic aromatic compounds, verify their significance in distinguishing different origins of RGM herbs and provide a simple and effective quality evaluation method. A selective extraction method was developed for noniridoid compounds and the extract was then subjected to UPLC-QTOF-MS analysis to obtain the RGM-MS ion pair database for noniridoid components. An HPLC-DAD quantitative analysis method was further developed based on characteristic aromatic compounds (2-phenylethyl ß-d-glucopyranoside, 2-methoxyanofinic acid and gentioxepine) after the ion screening in the MS database. By means of principal component analysis and hierarchical clustering analysis analysis, the significant relationship between aromatic compounds contents and different species of RGM was revealed. As a result, the significance of 2-phenylethyl ß-d-glucopyranoside, 2-methoxyanofinic acid and gentioxepine in distinguishing four species of RGM herbs was verified and a sensitive and reproducible HPLC-DAD method was established using these markers, which can be used for the classification and quantitative analysis of RGMs.

Pharmacokinetic Analysis of Four Bioactive Iridoid and Secoiridoid Glycoside Components of Radix Gentianae Macrophyllae and Their Synergistic Excretion by HPLC-DAD Combined with Second-Order Calibration.

An HPLC-DAD method combined with second-order calibration based on the alternating trilinear decomposition (ATLD) algorithm with the aid of region selection was developed to simultaneously and quantitatively characterize the synergistic relationships and cumulative excretion of the four bioactive ingredients of Radix Gentianae Macrophyllae in vivo. Although the analytes spectra substantially overlapped with that of the biological matrix, the overlapping profiles between analytes and co-eluting interferences can be successfully separated and accurately quantified by the ATLD method on the basis of the strength of region selection. The proposed approach not only determined the content change but also revealed the synergistic relationships and the cumulative excretion in vivo of the four ingredients in urine and feces samples collected at different excretion time intervals. In addition, several statistical parameters were employed to evaluate the accuracy and precision of the method. Quantitative results were confirmed by HPLC-mass spectrometry. Satisfactory results indicated that the proposed approach can be utilized to investigate the pharmacokinetics of Radix Gentianae Macrophyllae excretion in vivo.

Studies on genetic diversity in endangered medicinal plant Radix Gentianae Macrophyllae of Gansu / 中华中医药杂志

Objective:To study genetic diversity in endangered medicinal plant Radix Gentianae Macrophyllae of Gansu,then to provide theoretical evidence for the sustainable development and reseanable use.Methods:The-20℃Radix Gentianae Macrophyllae iced leaves for enzyme extraction,electrophoresis,staining and analysis POPGENE software.Results:9 of Radix Gentianae Macrophyllae wild populations of 151 individuals through the eight enzyme systems and 12 enzyme loci analysis showed that lower levels of genetic diversity existed in Radix Gentianae Macrophyllae.Each point the average number of alleles locus(A)was 1.1,the percentage of polymorphic loci(P)was 20.17,and the GST was 0.1352,suggesting that 86.48%of the total genetic diversity was distributed within populations.Conclusion:Considering the relatively low levels of genetic deversity in Radix Gentianae Macrophyllae populations and the many seedlings regenerating in the smallest population,we recommend in situ conservation in Tianshui,Ziwuling to maintain viable Radix Gentianae Macrophyllae populations in their original habitats.

Determination of Gentiopicroside in Radix Gentianae Macrophyllae Produced in Gansu Province by HPLC / 中药新药与临床药理

Objective To determine the content of gentiopicroside and to evaluate the quality of Radix Gentianae Macrophyllae in Gansu province.Methods The HPLC method was performed on Waters C18 column.Methanol-water(3 ∶7) were used as the mobile phase,the flow rate was at 1.0 mL?min-1 with the detection wavelength being 254 nm.Results For gentiopicroside,the linear range was 1.7352～17.3520 ?g and the average recovery was 97.06 %with RSD=2.5 %.Conclusion The content of gentiopicroside in all of the samples produced in Gansu province is higher than or approach to the standard for Radix Gentianae Macrophyllae in Chinese Pharmacopoeia.