LC-MS/MS analysis and evaluation of the anti-inflammatory activity of components from BushenHuoxue decoction.
BACKGROUND
BushenHuoxue decoction (BSHXD) is a Chinese medicine prescription, which is composed of nine Chinese medical materials, used to treat osteoarthritis (OA).


OBJECTIVE
This study develops sensitive and convenient LC-MS/MS methods to analyze chemical components from BSHXD, and assess the anti-inflammatory activities thereof.


METHODS
The chemical composition from BSHXD water extract was qualitative analyzed by high-performance liquid chromatography coupled with electrospray ionization quadrupole time-of-flight mass spectrometry (HPLC-ESI-Q-TOF-MS). Twelve reference compounds were analyzed by UPLC-ESI-MS/MS. Anti-inflammatory activities of target components were assessed by ELISA at 20 and 100 μg/mL.


RESULTS
It is the first time that 88 compounds were qualitatively identified from BSHXD, of which 12 with potential in treating OA according to the literature were quantified. Within BSHXD the contents of quercetin, isopsoralen, icarisideII, osthole, and isoimperatorin increased remarkably compared with those in single herb which make up BSHXD, the contents were 0.1999, 0.4634, 0.0928, 0.5364, and 0.1487 mg/g. ELISA data displayed that BSHXD and the five compounds mentioned inhibited the expressions of TNF-α, IL-6 and NO released from LPS-stimulated RAW264.7 cell, with maximum inhibition rates of 104.05% (osthole, 100 μg/mL), 100.03% (osthole, 100 μg/mL), and 93.46% (isopsoralen, 20 μg/mL), respectively.


CONCLUSIONS
Content changes of 12 compounds in BSHXD and single herbs which comprise the prescription were measured and analyzed. Contents of five compounds increased may be explained by solubilization between drugs and chemical reaction. ELISA results reported that the increased contents of the five compounds could inhibit expression of the inflammatory factors.
a Translational Medicine Center of Nanjing University of Chinese Medicine , Nanjing , China.|b Jiangsu Collaborative Innovation Center of Traditional Chinese Medicine (TCM) Prevention and Treatment of Tumor , Nanjing , China.
Dongdong
D
Sun
a Translational Medicine Center of Nanjing University of Chinese Medicine , Nanjing , China.|c Key Laboratory of Famous Doctors' Proved Recipe Evaluation and Transformation of State Administration of Traditional Chinese Medicine , Nanjing , China.
Qiuying
Q
Yan
b Jiangsu Collaborative Innovation Center of Traditional Chinese Medicine (TCM) Prevention and Treatment of Tumor , Nanjing , China.
Xiaofang
X
Xu
a Translational Medicine Center of Nanjing University of Chinese Medicine , Nanjing , China.|c Key Laboratory of Famous Doctors' Proved Recipe Evaluation and Transformation of State Administration of Traditional Chinese Medicine , Nanjing , China.
Weixing
W
Shen
a Translational Medicine Center of Nanjing University of Chinese Medicine , Nanjing , China.|c Key Laboratory of Famous Doctors' Proved Recipe Evaluation and Transformation of State Administration of Traditional Chinese Medicine , Nanjing , China.
Changliang
C
Xu
a Translational Medicine Center of Nanjing University of Chinese Medicine , Nanjing , China.|c Key Laboratory of Famous Doctors' Proved Recipe Evaluation and Transformation of State Administration of Traditional Chinese Medicine , Nanjing , China.
Jiani
J
Tan
b Jiangsu Collaborative Innovation Center of Traditional Chinese Medicine (TCM) Prevention and Treatment of Tumor , Nanjing , China.
Haibin
H
Zhang
b Jiangsu Collaborative Innovation Center of Traditional Chinese Medicine (TCM) Prevention and Treatment of Tumor , Nanjing , China.
Liu
L
Li
a Translational Medicine Center of Nanjing University of Chinese Medicine , Nanjing , China.|b Jiangsu Collaborative Innovation Center of Traditional Chinese Medicine (TCM) Prevention and Treatment of Tumor , Nanjing , China.
Haibo
H
Cheng
D000818:Animals; D000893:Anti-Inflammatory Agents / Q000032:analysis / Q000494:pharmacology*; D002138:Calibration; D002851:Chromatography, High Pressure Liquid / Q000592:standards; D004365:Drugs, Chinese Herbal / Q000032:analysis / Q000494:pharmacology*; D004797:Enzyme-Linked Immunosorbent Assay; D018836:Inflammation Mediators / Q000378:metabolism; D015850:Interleukin-6 / Q000378:metabolism; D016014:Linear Models; D008070:Lipopolysaccharides / Q000494:pharmacology; D008264:Macrophages / Q000187:drug effects* / Q000378:metabolism; D051379:Mice; D009569:Nitric Oxide / Q000378:metabolism; D000067996:RAW 264.7 Cells; D012015:Reference Standards; D021241:Spectrometry, Mass, Electrospray Ionization / Q000592:standards; D053719:Tandem Mass Spectrometry / Q000592:standards; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism
D000893:Anti-Inflammatory Agents; D004365:Drugs, Chinese Herbal; D018836:Inflammation Mediators; D015850:Interleukin-6; D008070:Lipopolysaccharides; D014409:Tumor Necrosis Factor-alpha; C527851:bushen huoxue; C577686:interleukin-6, mouse; D009569:Nitric Oxide
ELISA; osteoarthritis; qualitative analysis; simultaneously determine