Herbal Formula Science (대한한의학방제학회지)

The Korean Medicine Society for the Herbal Formula Study (대한한의학방제학회)
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LC-MS/MS analysis and anti-inflammatory effects of crude extract from Coptidis Rhizoma

황련 추출물의 LC-MS/MS 분석 및 항염증 효과

  • Min-Jung, Kim (College of Veterinary Medicine, Gyeongsang National University) ;
  • Ye-Jin, Yang (College of Veterinary Medicine, Gyeongsang National University) ;
  • Kwang-Youn, Kim (Korean Medicine (KM) Application Center, Korea Institute of Oriental Medicine) ;
  • Hun Hwan, Kim (College of Veterinary Medicine, Gyeongsang National University) ;
  • Jae Dong, Son (College of Veterinary Medicine, Gyeongsang National University) ;
  • Ju-Hye, Yang (Korean Medicine (KM) Application Center, Korea Institute of Oriental Medicine) ;
  • Dong bin, Lee (College of Veterinary Medicine, Gyeongsang National University) ;
  • Woo Hyun, Kim (College of Veterinary Medicine, Gyeongsang National University) ;
  • Hu-Jang, Lee (College of Veterinary Medicine, Gyeongsang National University) ;
  • Seon Been, Bak (School of Korean Medicine, Dongguk University) ;
  • Kwang-Il, Park (College of Veterinary Medicine, Gyeongsang National University)
  • 김민정 (경상국립대학교 수의과대학) ;
  • 양예진 (경상국립대학교 수의과대학) ;
  • 김광연 (한국한의학연구원 한의기술응용센터) ;
  • 김훈환 (경상국립대학교 수의과대학) ;
  • 손재동 (경상국립대학교 수의과대학) ;
  • 양주혜 (한국한의학연구원 한의기술응용센터) ;
  • 이동빈 (경상국립대학교 수의과대학) ;
  • 김우현 (경상국립대학교 수의과대학) ;
  • 이후장 (경상국립대학교 수의과대학) ;
  • 박선빈 (동국대학교 한의과대학 방제학교실) ;
  • 박광일 (경상국립대학교 수의과대학)
  • Received : 2023.01.09
  • Accepted : 2023.01.12
  • Published : 2023.02.28
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Abstract

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 flow rate of 0.5 mL/min for analysis, and a Prontosil C18 column (length, 250 mm; inner diameter, 4.6 mm; particle size, 5 ㎛; 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 ㎍/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.

Keywords

Acknowledgement

This work was supported by the National Research Foundation (NRF) grant funded by the Korean government (No. 2022R1I1A3053818), the "Leaders in Industry-university Cooperation 3.0" Project, and supported by the Ministry of Education and National Research Foundation of Korea and by grant (No. 20190055) funded by the Ministry of Oceans and Fisheries, Republic of Korea.

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