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KMID : 1149220230310010001
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Korean Journal of Oriental Medical Prescription
2023 Volume.31 No. 1 p.1 ~ p.10
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LC-MS/MS analysis and anti-inflammatory effects of crude extract from Coptidis Rhizoma
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Kim Min-Jung
Yang Ye-Jin
Kim Kwang-Youn
Kim Hun-Hwan
Son Jae-Dong
Yang Ju-Hye
Lee Dong-Bin
Kim Woo-Hyun
Lee Hu-Jang
Park Seon-Been
Park Kwang-Il
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Abstract
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Objectives : The main aim of this study was to examine the LC-MS/MS used to identify phenolic compounds of CRE(Coptidis Rhizoma 70% EtOH Extract). Also, we investigated antioxidative activities and Anti-inflammatory activities.
Methods : LC-MS/MS Analysis HPLC and LC-MS/MS were performed on a 1260 series HPLC system (Agilent Technologies, Inc., California, USA) and 3200 QTrap tandem mass system (Sciex LLC) operated in positive ion mode (spray voltage set at ?4.5 kV). The solvent used was DW and Acetonitrile containing 0.1% formic acid, a gradient system was used at a ?ow rate of 0.5 mL/min for analysis, and a Prontosil C18 column (length, 250 mm; inner diameter, 4.6 mm; particle size, 5 μm; Phenomenex Co., Ltd., California, USA, Biochoff Chromatography) was used. The solvent conditions used in the mobile phases were 0?10 min at 10?15% B, 10?20 min at 20% B, 20?30 min at 25%, 30?40 min at 40%, 40?50 min at 70%, 50?60 min at 95%, and 60?70 min at 95%. The analysis was performed at a wavelength of 284 nm and a temperature of 35℃. The cell viability was measured using a 3-(4,5-dimethyethiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity. We examined the effects of CRE on the lipopolysaccharide (LPS)-induced production of nitric oxide (NO) in a RAW 264.7 cells
Results : The chemical analysis CRE by Liquid chromatography-tandem mass spectrometry (LC-MS/MS) confirmed that Rosmarinic acid, Ferrulic acid, 3-O-feruloylquinic acid, and 5-O-feruloylquinic acid as phenolic components. DPPH radical scavenging activity was the inhibitory activity of CRE showed at 200 μg/mL a statistically significant level. MTT assay demonstrated that the CRE did not have a cytotoxic effect in RAW 264.7 and LPS-induced RAW264.7 cells. Also, CRE reduced NO production in RAW 264.7 cells stimulated with LPS.
Conclusions : Based on these findings, The chemical analysis 4 major components CRE such as Rosmarinic acid, Ferrulic acid, 3-O-feruloylquinic acid, and 5-O-feruloylquinic acid. Moreover, we confirmed that CRE has effects antioxidant and anti-inflammatory. The results demonstrate that CRE can be used as an antioxidant and a powerful chemopreventive ingredient against inflammatory diseases.
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KEYWORD
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LC-MS/MS, Anti-inflammatory, Antioxidant, Coptidis Rhizoma
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