Jing Qi, Zhang Min, Yao Yi-Qing, Wang Lian-Xing, Tian Xiao-Jin, Xu Xiao-Yan, Wang Hong-da, Chen Qin-Hua, Liu Er-Wei, Gao Xiu-Mei, Yang Wen-Zhi
State Key Laboratory of Chinese Medicine Modernization, Tianjin University of Traditional Chinese Medicine, 10 Poyanghu Road, Tianjin 301617, China; Haihe Laboratory of Modern Chinese Medicine, 10 Poyanghu Road, Tianjin 301617, China.
Shenzhen Baoan Authentic TCM Therapy Hospital, Shenzhen 518101, China.
J Pharm Biomed Anal. 2025 Nov 15;265:117041. doi: 10.1016/j.jpba.2025.117041. Epub 2025 Jun 26.
Erzhi formula (EZF) consists of wine-processed Ligustri Lucidi Fructus (W-LLF) and Ecliptae Herba (EH), while Jiawei Erzhi formula (JWEZF) is derived by newly incorporating Spatholobi Caulis (SC) and Achyranthis Bidentatae Radix (ABR). To date, the chemical difference between EZF and JWEZF has been seldom investigated. This work was designed to qualitatively and quantitatively evaluate the chemical difference between JWEZF and EZF by dimension-enhanced liquid chromatography-mass spectrometry approaches. An on-line comprehensive two-dimensional liquid chromatography/quadrupole time-of-flight mass spectrometry (2D-LC/QTOF-MS) approach was developed for the multi-component characterization of both EZF and JWEZF, while scheduled multiple reaction monitoring (sMRM) was employed for quantitative assay of the potentially differential components. Especially, the vivid 2D-LC contour plots and 4D data plots were utilized to embody the holistic chemical differentiation. A total of 289 compounds in JWEZF and 225 compounds in EZF were identified or tentatively characterized (e.g. iridoids, flavonoids, triterpenoids, phenolic acids, alkaloids, and organic acids). Compared to EZF, we could newly characterize 52 compounds in JWEZF which originated from SC and ABR, together with 12 newly generated compounds. The quantitative assay experiment determined the contents of six compounds exclusively found in JWEZF (catechin, epicatechin, chikusetsusaponin IVa, ginsenoside Ro, achyranthoside E, and β-ecdysterone), and validated the increased contents of 11 compounds in JWEZF that were common for these two formulas. The co-decoction of W-LLF with SC/ABR might cause the observed content variation for these 11 compounds between EZF and JWEZF. The developed dimension-enhanced approaches can better decipher the chemical difference between different TCM compound formulas.
二至丸（EZF）由酒制女贞子（W-LLF）和墨旱莲（EH）组成，而加味二至丸（JWEZF）是在二至丸的基础上新增了鸡血藤（SC）和牛膝（ABR）。迄今为止，很少有研究探讨EZF和JWEZF之间的化学差异。本研究旨在通过维度增强液相色谱-质谱联用方法对JWEZF和EZF之间的化学差异进行定性和定量评估。开发了一种在线全二维液相色谱/四极杆飞行时间质谱（2D-LC/QTOF-MS）方法用于同时对EZF和JWEZF进行多成分表征，同时采用定时多反应监测（sMRM）对潜在的差异成分进行定量分析。特别地，利用直观的二维液相色谱等高线图和四维数据图来体现整体化学差异。共鉴定或初步表征了JWEZF中的289种化合物和EZF中的225种化合物（如环烯醚萜类、黄酮类、三萜类、酚酸类、生物碱类和有机酸类）。与EZF相比，在JWEZF中新增鉴定出52种源自SC和ABR的化合物，以及12种新生成的化合物。定量分析实验测定了仅在JWEZF中发现的6种化合物（儿茶素、表儿茶素、竹节人参皂苷IVa、人参皂苷Ro、牛膝甾苷E和β-蜕皮甾酮）的含量，并验证了JWEZF中11种在两个方剂中均存在的化合物含量增加。W-LLF与SC/ABR的共煎可能导致这11种化合物在EZF和JWEZF之间出现含量差异。所开发的维度增强方法能够更好地解读不同中药复方之间的化学差异。
