Identification of Chemical Constituents in Coptis chinensis Inflorescence by UPLC-Q-Orbitrap HRMS / 中国实验方剂学杂志

Objective:An ultra-high performance liquid chromatography coupled with quadrupole-orbitrap high resolution mass spectrometry （UPLC-Q-Orbitrap HRMS） was developed to analyze and identify the chemical constituents in <italic>Coptis chinensis</italic> inflorescence. Method:The chromatographic separation was performed on ACQUITY UPLC BEH C₁₈ column （2.1 mm×100 mm， 1.7 μm） with the mobile phase of 0.1% formic acid aqueous solution （A）-acetonitrile （B） for gradient elution （0-15 min， 10%-22%B； 15-20 min， 22%B； 20-25 min， 22%-44%B； 25-35 min， 44%-50%B； 35-40 min， 50%-60%B； 40-55 min， 60%-85%B）， the flow rate was 0.15 mL·min^-1， the injection volume was 3 μL and the column temperature was 30 ℃. HRMS was equipped with electrospray ionization （ESI） and scanned in positive and negative ion modes by means of full scan/data dependent secondary scan （Full MS/dd-MS²）. Compound Discoverer 3.0 software combined with mzCloud， mzVault， ChemSpider databases and HRMS database of components in traditional Chinese medicine were used to analyze and identify the collected data by HRMS， based on accurate relative molecular mass， retention time and characteristic ion fragmentation of the compounds， as well as literature information and relevant reference materials. Result:A total of 51 chemical constituents were identified in <italic>C</italic>.<italic> chinensis</italic> inflorescence， including 16 alkaloids， 14 flavonoids， 7 phenylpropanoids， 7 organic acids and 7 others. Among them， 10 components ［berberine， palmatine， coptidine， rutin， quercetin， isoquercitrin， chlorogenic acid， cryptochlorogenic acid，<italic> D</italic>-（-） quinic acid and <italic>D</italic>-proline］ were unambiguously identified by comparing with reference standards. Conclusion:The established UPLC-Q-Orbitrap HRMS can be used to accurately analyze and identify chemical constituents of <italic>C. chinensis</italic> inflorescence. A total of 41 chemical constituents are reported from <italic>C. chinensis</italic> inflorescence for the first time and 6 alkaloids are found from the <italic>C. chinensis</italic> for the first time. These findings can provide methodological reference and experimental basis for the basic research of quality evaluation and efficacy materials of <italic>C. chinensis</italic> inflorescence， and lay a foundation for its further development and utilization.

Establishment of HPLC Fingerprint of Coptis chinensis Inflorescence and Its Spectrum-effect Relationship with Antioxidant and Antibacterial Effects / 中国药房

OBJECTIVE：To establish HPLC fingerprint of Coptis chinensis inflorescence，and study its spectrum-effect relationship with antioxidant and antibacterial effects. METHODS ：Taking 14 batches of C. chinensis inflorescence from different producing areas as the object ，HPLC method was adopted. The determination was performed on Supersil C 18 column with mobile phase consisted of acetonitrile- 0.1％ phosphoric acid solution（gradient elution ）at the flow rate of 1.0 mL/min. The column temperature was set at 25 ℃. The detection wavelength was set at 329 nm，and sample size was 10 μL. The fingerprints of 14 batches of C. chinensis inflorescence were established by Similarity Evaluation System of TCM Fingerprint （2012 A edition ），and the similarity evaluation and common peak identification were carried out. Taking DPPH free radical scavenging rate and hydroxyl radical scavenging rate as antioxidant effects index ，relative antibacterial activity （Escherichia coli ）as antibacterial effect index ， SPSS 21.0 software was adopted to analyze the Pearson correlation between common peaks of C. chinensis inflorescence and above efficacy indexes ；their spectrum-effect relationship was established and validated. RESULTS ：A total of 7 common peaks were obtained in HPLC fingerprint of C. chinensis inflorescence，and the similarity was no less than 0.916. No. 5 peak was identified as berberine hydrochloride. Seven common peaks were positively correlated with DPPH free radical scavenging rate ；No. 1-3，4，6，7 peaks were positively correlated with hydroxyl radical scavenging rate ，while No. 5 peak was negatively correlated with hydroxyl radical scavenging rate. There was a positive correlation between No. 5 peak and antibacterial activity in vitro . After validation ， relative error between the predicted values and the measured values of DPPH free radical scavenging rate ，hydroxyl radical scavenging rate and relative antibacterial activity was 0.92％- 14.5％ . CONLUSIONS ：The established spectrum-effect relationship model can be used to evaluate antioxidant andantibacterial effects of C. chinensis inflorescence. The chemical components represented by No. 1，2，3，4，6，7 peaks are the material basis of antioxidant effect of C. chinensis inflorescence， and berberine hydrochloride is the material basis of antibacterial effect.