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《Chinese Journal of Pharmaceutical Analysis》 2014-02
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Identification and simultaneous determination of caffeoylquinic acids in Huahong capsules

SHEN Shou-mao;SHEN Lian-gang;ZHANG Jing;LI Guang-zhi;ZHANG Yan-hua;LIU Chun-ming;SI Jian-yong1#;Institute of Medicinal Plant Development,Chinese Academy of Medical Sciences,Peking Union Medical College;School of Pharmacy,Yancheng Teachers' University;Guangxi Huahong Pharmaceutical Co. Ltd.;  
Objective: To analyze the caffeoylquinic acids and to develop an HPLC method for simultaneous determination of neochlorogenic acid,chlorogenic acid and cryptochlorogenin acid in Huahong capsules. Methods: A UPLC / Q- TOF- MS / MS method was developed to rapidly identify the caffeoylquinic acids in Huahong capsules. And the UPLC separation was performed on a Thermo BEH C 18 column( 2. 1 mm × 100 mm,1. 7 μm) with 0. 1% formic acid acetonitrile- 0. 1% formic acid as the mobile phase in gradient elution at a flow rate of 0. 4 μL · min- 1. The column temperature was set at 40 °C. The MS spectrometry detection was performed with electrospray ionization in negative ion mode,and using the following operating parameters: scan range,100- 1500 m / z; capillary voltage,2. 5 kV; skimmer voltage,40 kV; ion source temperature,120 °C; drying gas temperature,450 °C and sample collision energy,15- 40 eV. Besides,HPLC was performed on a Phenomenex C18( 250 mm ×4. 6 mm,4 μm)column with 0. 1% formic acid acetonitrile- 0. 1% formic acid as the mobile phase in gradient elution( 0- 7 min,5% →8% A; 7- 17 min,8% →15% A; 17- 45 min,15% →30% A; 45- 55 min,30% →50% A; 55- 60 min,50% →100% A) at a flow rate of 1. 0 mL·min- 1. The detect wavelength was set at 326 nm,and the column temperature was set at 30 °C. Results: Based on the UPLC / Q- TOF- MS / MS analysis,four caffeoylquinic acids,neochlorogenic acid,chlorogenic acid and cryptochlorogenin acid and isochlorogenic acid A,were identified from Huahong capsules. In addition,by using HPLC method,the correlation between the concentration and the chromatographic peak area of neochlorogenic acid,chlorogenic acid and cryptochlorogenin acid was linear in ranges of 3. 54-176. 80 μg·mL- 1( r = 0. 9999),6. 98- 348. 80 μg·mL- 1( r= 0. 9999),4. 06- 202. 80 μg·mL- 1( r =0. 9999),respectively. The average recoveries of the three caffeoylquinic acids were in the range of 101. 5%-102. 2%,and the relative standard deviations( RSD) were in the range of 1. 9%- 2. 4%. Conclusion: The UPLC /Q- TOF- MS / MS and HPLC methods do not only pave a way for further understanding of the chemical constituents in Huahong capsules,but also provide scientific evidences for the quality control of Huahong capsules.
【Fund】: 国家“重大新药创制”科技专项(Nos.2011ZX09307-002-01 2012ZX09301-002-001)
【CateGory Index】: R284.1
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