滇龙胆FTIR&HPLC特征指纹图谱表征及多元评价体系研究

Natural products in traditional Chinese medicines (TCM) are complex and diverse, which need sensitive and reliable analytical methods to control the quality of product entirely, and ensure the clinical application safe and effective. TCM fingerprint is the foundation of the construction of modern Chinese medicine quality standard system, which provides an opportunity for systemic and comprehensive evaluation of TCM quality. Gentiana rigescens is the genuine medicinal material in Yunnan. Currently, the main problems in the quality evaluation of Gentiana rigescens are that evaluation method is not systematic and sampling is not representative. In this study, we will collect the resource of Gentiana rigescens in southwestern China, apply Fourier transform infrared spectroscopy (FTIR) and high performance liquid chromatography (HPLC), and combine chemometrics to (1) build FTIR and HPLC characteristic fingerprints and characterize their consistency, in order to construct the qualitative discrimination model of multidimensional fingerprint; (2) screen the characteristic peaks of FTIR spectra, with the help of multivariate statistics analysis method, the FTIR quantitative model of effective constituents will be set up based on the fitting of FTIR spectra and the contents of effective constituents; (3) multiple evaluation system will be integrated by confluence analysis of qualitative discriminant model and quantitative model. The system could be used to identify Gentiana rigescens effectively, evaluate the quality systematically in the chemical composition level, clarify the characteristics of stoichiometirc, discus the excellent provenance selection model, and provide theoretical basis for quality standardization of Gentiana rigescens.

中药天然产物复杂多样，亟需灵敏、可靠的分析方法整体控制产品质量，保证临床用药的安全有效。中药指纹图谱是现代中药质量标准体系建构的基础，为系统、全面评价中药质量提供契机。滇龙胆为云南道地药材，目前其质量评价主要存在评价方法缺乏系统性、取样不具有代表性等问题。本项目拟收集我国西南地区滇龙胆资源，采用傅里叶变换红外光谱（FTIR）和高效液相色谱（HPLC），结合化学计量学：（1）构建FTIR和HPLC特征指纹图谱，对其一致性进行表征，建立滇龙胆多维指纹图谱定性判别模型；（2）筛选FTIR光谱特征峰，利用多元统计分析方法将FTIR光谱与有效成分含量相拟合，建立有效成分的FTIR定量模型；（3）将定性判别模型与定量模型整合分析，集成滇龙胆多元评价体系。该体系可有效鉴别滇龙胆，在化学成分层面系统评价其质量，阐明种质资源化学计量特征，探讨优质种源筛选模型，为滇龙胆药材质量标准化提供理论依据。

滇龙胆（Gentiana rigescens Franch.）为龙胆科（Gentianaceae）多年生草本植物，其干燥根及根茎为中药龙胆来源之一，为云南道地药材，具有清热燥湿、泻肝胆火的功效。滇龙胆质量评价研究存在的主要问题有：（1）质量评价方法单一，缺乏系统性；（2）我国西南地区地理环境复杂、气候多变，不同产地药材质量参差不齐，取样缺乏代表性。为保证滇龙胆药材质量的一致性和稳定性，本项目以我国西南地区野生和栽培滇龙胆为研究对象，采用傅里叶变换红外光谱（FTIR）和高效液相色谱（HPLC）结合化学计量学方法，构建了不同产地、部位和生长年限的滇龙胆FTIR定性判别模型，以及有效成分含量的FTIR快速预测定量模型。研究结果显示滇龙胆产区间、部位间和部位内主要化学成分差异显著，表明滇龙胆的质量高度依赖地理环境。对HPLC和FTIR指纹图谱的一致性和相关性进行表征，HPLC和FTIR指纹图谱都显示GA-PLS-DA方法建立的模型是最稳健的识别滇龙胆产地的模型，说明HPLC和FTIR具有一致性。HPLC-FTIR二维异质相关分析结果显示，HPLC和FTIR具有物质相关性，说明光谱相关色谱用于多组分比较是可行的，就滇龙胆的定性识别而言，利用FTIR较HPLC更加方便，快捷，环境友好及费用低。通过项目的实施，集成滇龙胆定性判别与定量分析为一体的多元评价体系，为中药材的质量控制提供一种综合的参考方法，相关研究结果发表SCI论文8篇。项目实施期间，2人晋升副高级职称。

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