HPLC-DAD法同时测定清开灵注射液中7个酚酸类成分

目的: 建立同时测定清开灵注射液中7个酚酸类成分(新绿原酸、绿原酸、隐绿原酸、咖啡酸、3,4-二咖啡酰奎宁酸、3,5-二咖啡酰奎宁酸、4,5-二咖啡酰奎宁酸)含量的方法。方法: 采用Agilent Zorbax SB-C₁₈色谱柱(250 mm×4.6 mm,5 μm),流动相为0.1%甲酸水溶液(A)-乙腈(B),梯度洗脱,流速为0.5 mL·min^-1,检测波长327 nm,柱温30 ℃。结果: 80 min内7个待测组分分离度良好,在各自的检测范围内与峰面积呈良好的线性,其相关系数均大于0.9998,加样回收率为96.1%～103.9%。应用所建立的方法同时测定了清开灵注射液中新绿原酸、绿原酸、隐绿原酸、咖啡酸、4,5-二咖啡酰奎宁酸、3,4-二咖啡酰奎宁酸、3,5-二咖啡酰奎宁酸共7个成分的含量,并对5个不同批次的注射液进行了含量测定,其结果有一定的差异。结论: 本法可为清开灵注射液中酚酸类成分的质量控制提供参考。

Objective: To establish an HPLC-DAD method for simultaneous determination of 7 phenolic acids (neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, caffeic acid, 3, 4-dicaffeoylquinic acid, 3, 5-dicaffeoylquinic acid and 4, 5-dicaffeoylquinic acid) in Qingkailing injection. Methods: The separation was achieved on an Agilent Zorbax SB-C₁₈ analytical column (250 mm×4.6 mm, 5 μm) with a gradient elution of 0.1% formic acid(A) and acetonitrile(B) at a flow rate of 0.5 mL·min^-1. The detection wavelength was 327 nm and the column temperature was 30 ℃. Results: Perfect chromatogram with good separation was obtained within 80 min. All the seven calibration curves showed good linearity between concentration of each compound and peak area. And the value of r was higher than 0.9998 for all the analytes. The recoveries were 96.1%-103.9%. The developed method was successfully applied for the simultaneous determination of the 7 components (neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, caffeic acid, 4, 5-dicaffeoylquinic acid, 3, 4-dicaffeoylquinic acid and 3, 5-dicaffeoylquinic acid) in Qingkailing injection of five different batches which had great variation on the determination results. Conclusion: The developed method can provide reference for quality control of phenolic acids in Qingkailing injection.

1 ChP(中国药典). 2010. Vol Ⅰ(一部): 1110
2 CAO Jin(曹进), XU Yan(徐燕), WANG Yi-ming(王义明), et al. Determination of 3 components in Qingkailing injection by HPLC under different UV-vis waves(多波长高效液相色谱法测定清开灵注射液中3种有效成分). Chin J Pharm Anal(药物分析杂志), 2004, 24(1): 8
3 GAO Xiao-yan(高晓燕), LI Na(李娜), FAN Qiang (范强), et al. Study on the method of rapid quality control of aqueous solution of Flos Lonicerae Japonicae extraction in the preparation of Qingkailing injection(清开灵注射液生产过程中金银花提取液的在线质量控制方法研究). Spectrosc Spect Anal(光谱学与光谱分析), 2006, 26(5): 904
4 WU Yan-shu(武彦舒), JIN Cheng(金城),ZHANG Qian(张倩), et al. Simultaneous determination of ten active components in Qingkailing injection and relationship with quality control(清开灵注射液中10种有效成分的同时测定及其质量相关性研究). Spectrosc Spect Anal (光谱学与光谱分析), 2009, 29(11): 3112
5 XING Li-hong(邢丽红), LI Wen-long(李文龙), QU Hai-bin(瞿海斌). Rapid determination of 5 kinds of organic acid in Flos Lonicerae extracts by UV spectroscopy (金银花提取物中5种有机酸含量测定的紫外光谱法). Chin J Pharm Anal(药物分析杂志), 2011, 31(3): 547
6 NIU Xiao-xue(牛晓雪), CUI Xu-sheng(崔旭盛), SU He(苏贺) , et al. Simultaneous determination of seven compounds in Lonicera japonica by high performance liquid chromatography (高效液相色谱法同时测定忍冬中7 种成分). Chin J Chromatogr(色谱), 2012, 30(2): 211
7 LUO Qi-zhi(罗奇志), WANG You-zhi(王有志), LUO Jia-bo(罗佳波). HPLC-ESI/MS analysis of chlorogenic acid and its hydrolysate (绿原酸水解产物的高效液相色谱-电喷雾串联质谱分析). Chin J Pharm Anal(药物分析杂志), 2011, 31(7): 1345
8 Ma Yuan-chun, Wang Xiao-qian, Hou Fei-fei,et al. Rapid resolution liquid chromatography(RRLC) analysis and studies on the stability of Shuang-Huang-Lian preparations. J Pharm Biomed Anal, 2011, 54(2): 265
9 GU Li-hong(顾丽红), ZHU Pin-ye(朱品业). Influence of light and temperature on the stability of the solution of chlorogenic acid(日光和温度对绿原酸供试液稳定性的影响). Chin Tradit Pat Med(中成药), 1999, 21(11): 568
10 CHEN Gang(陈钢), HOU Shi-xiang(侯世祥), HU Ping(胡平), et al. Studies on stability of chlorogenic acid in extract of Flos Lonicerae(金银花提取物中绿原酸的稳定性研究). China J Chin Mater Med(中国中药杂志), 2003, 28(3): 223
11 LIU Yi(刘意), ZENG Gui-xian(曾桂先), SONG Feng-lan(宋凤兰), et al. Study on stability of chlorogenic acid(绿原酸稳定性研究). Chem Intermediates(化工中间体), 2009, 5(2): 25