title: Quantitative profiling of polar metabolites in herbal medicine injections for multivariate statistical evaluation based on independence principal component analysis.
abstract: Botanical primary metabolites extensively exist in herbal medicine injections (HMIs), but often were ignored to control. With the limitation of bias towards hydrophilic substances, the primary metabolites with strong polarity, such as saccharides, amino acids and organic acids, are usually difficult to detect by the routinely applied reversed-phase chromatographic fingerprint technology. In this study, a proton nuclear magnetic resonance (1H NMR) profiling method was developed for efficient identification and quantification of small polar molecules, mostly primary metabolites in HMIs. A commonly used medicine, Danhong injection (DHI), was employed as a model. With the developed method, 23 primary metabolites together with 7 polyphenolic acids were simultaneously identified, of which 13 metabolites with fully separated proton signals were quantified and employed for further multivariate quality control assay. The quantitative 1H NMR method was validated with good linearity, precision, repeatability, stability and accuracy. Based on independence principal component analysis (IPCA), the contents of 13 metabolites were characterized and dimensionally reduced into the first two independence principal components (IPCs). IPC1 and IPC2 were then used to calculate the upper control limits (with 99% confidence ellipsoids) of χ2 and Hotelling T2 control charts. Through the constructed upper control limits, the proposed method was successfully applied to 36 batches of DHI to examine the out-of control sample with the perturbed levels of succinate, malonate, glucose, fructose, salvianic acid and protocatechuic aldehyde. The integrated strategy has provided a reliable approach to identify and quantify multiple polar metabolites of DHI in one fingerprinting spectrum, and it has also assisted in the establishment of IPCA models for the multivariate statistical evaluation of HMIs.
authors.affiliation: Tianjin State Key Laboratory of Modern Chinese Medicine, Tianjin University of Traditional Chinese Medicine, Tianjin, China; Research and Development Center of TCM, Tianjin International Joint Academy of Biotechnology and Medicine, Tianjin, China.
authors.forename: Miaomiao
authors.initials: M
authors.lastname: Jiang
authors.affiliation: Tianjin State Key Laboratory of Modern Chinese Medicine, Tianjin University of Traditional Chinese Medicine, Tianjin, China; Research and Development Center of TCM, Tianjin International Joint Academy of Biotechnology and Medicine, Tianjin, China.
authors.forename: Yujiao
authors.initials: Y
authors.lastname: Jiao
authors.affiliation: Tianjin State Key Laboratory of Modern Chinese Medicine, Tianjin University of Traditional Chinese Medicine, Tianjin, China; Research and Development Center of TCM, Tianjin International Joint Academy of Biotechnology and Medicine, Tianjin, China.
authors.forename: Yuefei
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Tianjin State Key Laboratory of Modern Chinese Medicine, Tianjin University of Traditional Chinese Medicine, Tianjin, China; Research and Development Center of TCM, Tianjin International Joint Academy of Biotechnology and Medicine, Tianjin, China.
authors.forename: Lei
authors.initials: L
authors.lastname: Xu
authors.affiliation: Tianjin State Key Laboratory of Modern Chinese Medicine, Tianjin University of Traditional Chinese Medicine, Tianjin, China; Research and Development Center of TCM, Tianjin International Joint Academy of Biotechnology and Medicine, Tianjin, China.
authors.forename: Meng
authors.initials: M
authors.lastname: Wang
authors.affiliation: China Buchang Hospital for Cardio-Cerebral Vascular Disease, Xi'an, China.
authors.forename: Buchang
authors.initials: B
authors.lastname: Zhao
authors.affiliation: China Buchang Hospital for Cardio-Cerebral Vascular Disease, Xi'an, China.
authors.forename: Lifu
authors.initials: L
authors.lastname: Jia
authors.affiliation: China Buchang Hospital for Cardio-Cerebral Vascular Disease, Xi'an, China.
authors.forename: Hao
authors.initials: H
authors.lastname: Pan
authors.affiliation: Tianjin State Key Laboratory of Modern Chinese Medicine, Tianjin University of Traditional Chinese Medicine, Tianjin, China; Research and Development Center of TCM, Tianjin International Joint Academy of Biotechnology and Medicine, Tianjin, China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Zhu
authors.affiliation: Tianjin State Key Laboratory of Modern Chinese Medicine, Tianjin University of Traditional Chinese Medicine, Tianjin, China.
authors.forename: Xiumei
authors.initials: X
authors.lastname: Gao
mesh_terms: D004365:Drugs, Chinese Herbal / Q000737:chemistry* / Q000378:metabolism; D029001:Herbal Medicine; D007267:Injections; D009682:Magnetic Resonance Spectroscopy / Q000379:methods*; D055442:Metabolome; D015999:Multivariate Analysis; D010946:Plants, Medicinal / Q000737:chemistry* / Q000378:metabolism; D025341:Principal Component Analysis
chemical_list: D004365:Drugs, Chinese Herbal; C540581:danhong