Lu Jia-Xi1, Zhang Chun-Xia2, Hu Ying2, Zhang Meng-Han1, Wang Ya-Nan3, Qian Yue-Xin2, Yang Jing1, Yang Wen-Zhi4, Jiang Miao-Miao5, Guo De-An6. 1. Tianjin State Key Laboratory of Modern Chinese Medicine, Tianjin University of Traditional Chinese Medicine, 312 Anshanxi Road, Tianjin 300193, China. 2. Tianjin State Key Laboratory of Modern Chinese Medicine, Tianjin University of Traditional Chinese Medicine, 312 Anshanxi Road, Tianjin 300193, China; Tianjin Key Laboratory of TCM Chemistry and Analysis, Tianjin University of Traditional Chinese Medicine, 312 Anshanxi Road, Tianjin 300193, China. 3. Institute of Materia Medica, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing, China. 4. Tianjin State Key Laboratory of Modern Chinese Medicine, Tianjin University of Traditional Chinese Medicine, 312 Anshanxi Road, Tianjin 300193, China; Tianjin Key Laboratory of TCM Chemistry and Analysis, Tianjin University of Traditional Chinese Medicine, 312 Anshanxi Road, Tianjin 300193, China. Electronic address: ywz_0504@163.com. 5. Tianjin State Key Laboratory of Modern Chinese Medicine, Tianjin University of Traditional Chinese Medicine, 312 Anshanxi Road, Tianjin 300193, China; Tianjin Key Laboratory of TCM Chemistry and Analysis, Tianjin University of Traditional Chinese Medicine, 312 Anshanxi Road, Tianjin 300193, China. Electronic address: miaomiaojiang@tjutcm.edu.cn. 6. Tianjin State Key Laboratory of Modern Chinese Medicine, Tianjin University of Traditional Chinese Medicine, 312 Anshanxi Road, Tianjin 300193, China; Shanghai Research Center for Modernization of Traditional Chinese Medicine, National Engineering Laboratory for TCM Standardization Technology, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, 501 Haike Road, Shanghai 201203, China. Electronic address: gda5958@163.com.
Abstract
BACKGROUND: The florets of Carthamus tinctorius L. (safflower) serve as the source of a reputable herbal medicine targeting gynecological diseases. Conventional investigations regarding the quality control of safflower, however, mainly focused on the secondary metabolites with primary metabolites ignored. PURPOSE: To holistically evaluate the quality difference of safflower samples collected from five different producing regions by multiple chemical and biological approaches with both the primary and secondary metabolites considered. METHODS: A precursor ions list-triggered data-dependent MS2 approach was established by ultra-high performance liquid chromatography/Q-Orbitrap mass spectrometry (UHPLC/Q-Orbitrap MS) to comprehensively identify the secondary metabolites from safflower. Primary metabolites were identified by various 1D and 2D nuclear magnetic resonance (NMR) experiments. Similarity evaluation and quantitative assays of all the characterized primary metabolites and a quinochalcone C-glycoside (QCG) marker, hydroxysafflor yellow A (HSYA), were performed by quantitative 1H NMR (qNMR) using an external standard method. Multiple in vitro models with respect to the antioxidant, anti-platelet aggregation, and antioxidant stress injury effects, were assayed to determine the efficacy differences. RESULTS: Totally thirteen primary metabolites (including one nucleoside, two sugars, five organic alkali/acids, and five amino acids) and 135 secondary metabolites (97 QCGs and 38 flavonoids) could be identified or tentatively characterized from safflower. Good chemical consistency was observed between the commercial safflower samples and a standard safflower sample, with similarity varying in the range of 0.95‒0.99. The results from qNMR-oriented quantitative experiments (thirteen primary metabolites and HSYA) and biological assays indicated the quality of safflower samples from Xinjiang (XJ-2 and XJ-4), Hunan (HuN-1 and HuN-2), and Sichuan (SC), was comparable to the standard safflower sample. CONCLUSION: The integration of multiple chemical (using two analytical platforms, UHPLC/Q-Orbitrap MS and NMR) and biological (four in vitro models) approaches by determining both the primary and secondary metabolites demonstrated a powerful strategy that could facilitate the holistic quality evaluation of traditional Chinese medicine.
BACKGROUND: The florets of Carthamus tinctorius L. (safflower) serve as the source of a reputable herbal medicine targeting gynecological diseases. Conventional investigations regarding the quality control of safflower, however, mainly focused on the secondary metabolites with primary metabolites ignored. PURPOSE: To holistically evaluate the quality difference of safflower samples collected from five different producing regions by multiple chemical and biological approaches with both the primary and secondary metabolites considered. METHODS: A precursor ions list-triggered data-dependent MS2 approach was established by ultra-high performance liquid chromatography/Q-Orbitrap mass spectrometry (UHPLC/Q-Orbitrap MS) to comprehensively identify the secondary metabolites from safflower. Primary metabolites were identified by various 1D and 2D nuclear magnetic resonance (NMR) experiments. Similarity evaluation and quantitative assays of all the characterized primary metabolites and a quinochalcone C-glycoside (QCG) marker, hydroxysafflor yellow A (HSYA), were performed by quantitative 1H NMR (qNMR) using an external standard method. Multiple in vitro models with respect to the antioxidant, anti-platelet aggregation, and antioxidant stress injury effects, were assayed to determine the efficacy differences. RESULTS: Totally thirteen primary metabolites (including one nucleoside, two sugars, five organic alkali/acids, and five amino acids) and 135 secondary metabolites (97 QCGs and 38 flavonoids) could be identified or tentatively characterized from safflower. Good chemical consistency was observed between the commercial safflower samples and a standard safflower sample, with similarity varying in the range of 0.95‒0.99. The results from qNMR-oriented quantitative experiments (thirteen primary metabolites and HSYA) and biological assays indicated the quality of safflower samples from Xinjiang (XJ-2 and XJ-4), Hunan (HuN-1 and HuN-2), and Sichuan (SC), was comparable to the standard safflower sample. CONCLUSION: The integration of multiple chemical (using two analytical platforms, UHPLC/Q-Orbitrap MS and NMR) and biological (four in vitro models) approaches by determining both the primary and secondary metabolites demonstrated a powerful strategy that could facilitate the holistic quality evaluation of traditional Chinese medicine.