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炮制方法对地黄化学成分的影响:基于超高效液相色谱-线性离子阱-静电场轨道阱质谱分析

[Effect of processing method on chemical constituents of Rehmanniae Radix: based on UHPLC-LTQ-Orbitrap MS].

作者信息

Lu Xing-Mei, Zhong Ling-Yun, Wang Shuo, Deng Yan-Wen, Liu Hong, Chen Ming-Xia, Huang Yi, Tong Heng-Li

机构信息

Jiangxi University of Chinese Medicine Nanchang 330004,China.

Jianchangbang Pharmaceutical Co., Ltd. Fuzhou 344700, China Bei Jing Scrianen Pharmaceutical Co., Ltd. Beijing 100176, China.

出版信息

Zhongguo Zhong Yao Za Zhi. 2023 Jan;48(2):399-414. doi: 10.19540/j.cnki.cjcmm.20220816.301.

Abstract

This study aims to explore the chemical composition of Rehmanniae Radix braised with mild fire and compare the effect of processing method on the chemical composition of Rehmanniae Radix. To be specific, ultra-high performance liquid chromatography with linear ion trap-orbitrap mass spectrometry(UHPLC-LTQ-Orbitrap MS) was used to screen the chemical constituents of Rehmanniae Radix. The chemical constituents were identified based on the relative molecular weight and fragment ions, literature information, and Human Metabolome Database(HMDB). The ion peak area ratio of each component before and after processing was used as the index for the variation. SIMCA was employed to establish principal component analysis(PCA) and orthogonal partial least squares discriminant analysis(OPLS-DA) models of different processed products. According to the PCA plot, OPLS-DA plot, and VIP value, the differential components before and after the processing were screened out. The changes of the content of differential components with the processing method were analyzed. A total of 66 chemical components were identified: 57 of raw Rehmanniae Radix, 55 of steamed Rehmanniae Radix, 55 of wine-stewed Rehmanniae Radix, 51 of repeatedly steamed and sundried Rehmanniae Radix Praeparata, 62 of traditional bran-braised Rehmanniae Radix, and 63 of electric pot-braised Rehmanniae Radix. Among them, the 9 flavonoids of braised Rehmanniae Radix were from Citri Reticulatae Pericarpium. PCA suggested significant differences in the chemical composition of Rehmanniae Radix Praeparata prepared with different processing methods. OPLS-DA screened out 32 chemical components with VIP value >1 as the main differential components. Among the differential components, 9 were unique to braised Rehmanniae Radix(traditional bran-braised, electric pot-braised) and the degradation rate of the rest in braised(traditional bran-braised, electric pot-braised) or repeatedly steamed and sundried Rehmanniae Radix was higher than that in the steamed or wine-stewed products. The results indicated the chemical species and component content of Rehmanniae Radix changed significantly after the processing. The 32 components, such as rehmapicrogenin, martynoside, jionoside D, aeginetic acid, hesperidin, and naringin, were the most important compounds to distinguish different processed products of Rehmanniae Radix. The flavonoids introduced by Citri Reticulatae Pericarpium as excipient may be the important material basis for the effectiveness of braised Rehmanniae Radix compared with other processed products.

摘要

本研究旨在探究文火炖制熟地黄的化学成分,并比较炮制方法对熟地黄化学成分的影响。具体而言,采用超高效液相色谱-线性离子阱-轨道阱质谱联用技术(UHPLC-LTQ-Orbitrap MS)筛选熟地黄的化学成分。基于相对分子量、碎片离子、文献信息以及人类代谢组数据库(HMDB)对化学成分进行鉴定。将各成分炮制前后的离子峰面积比作为变化指标。运用SIMCA建立不同炮制品的主成分分析(PCA)模型和正交偏最小二乘法判别分析(OPLS-DA)模型。根据PCA图、OPLS-DA图及VIP值,筛选出炮制前后的差异成分。分析差异成分含量随炮制方法的变化情况。共鉴定出66种化学成分:生地黄57种、熟地黄55种、酒炖熟地黄55种、九蒸九晒熟地黄51种、传统麸炒熟地黄62种、电饭煲炖熟地黄63种。其中,炖制熟地黄中的9种黄酮类成分来自陈皮。PCA表明不同炮制方法制备的熟地黄在化学成分上存在显著差异。OPLS-DA筛选出VIP值>1的32种化学成分作为主要差异成分。在差异成分中,9种是炖制熟地黄(传统麸炒、电饭煲炖)所特有的,其余成分在炖制(传统麸炒、电饭煲炖)或九蒸九晒熟地黄中的降解率高于清蒸或酒炖制品。结果表明,熟地黄炮制后化学物种及成分含量发生了显著变化。地黄苦苷元、麦角硫因苷、梓醇苷D、京尼平酸、橙皮苷和柚皮苷等32种成分是区分不同熟地黄炮制品的最重要化合物。作为辅料的陈皮引入的黄酮类成分可能是炖制熟地黄与其他炮制品相比发挥药效的重要物质基础。

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