基于超高效液相色谱指纹图谱、多指标成分定量及化学计量学的小儿感冒颗粒质量评价  

Quality evaluation of Xiao'er Ganmao granules based on UPLC fingerprint,multi-component determination,and chemometric analysis

作　　者：李沁 张聪 陈亮 陈在敏 LI Qin;ZHANG Cong;CHEN Liang;CHEN Zai-min(Fujian Institute for Food and Drug Quality Control,Fuzhou 350001,China)

机构地区：[1]福建省食品药品质量检验研究院,福州350001

出　　处：《药物分析杂志》2023年第9期1612-1622,共11页Chinese Journal of Pharmaceutical Analysis

基　　金：国家药品监督管理局“中药有效性安全性评价及全过程质量控制研究”子课题(NMPAJGKX-2023-092);福建省药品监督管理局科技计划项目(2022008)。

摘　　要：目的:建立小儿感冒颗粒的超高效液相色谱(UPLC)指纹图谱、多指标成分含量测定和化学计量学相结合的质量评价方法。方法:采用Waters Acquity UPLC BEH C18色谱柱(100 mm×2.1 mm,1.7μm),以乙腈-0.2%磷酸水溶液为流动相,梯度洗脱,流速为0.3 mL·min-1,柱温为30℃,多波长切换检测。使用中药色谱指纹图谱相似度评价软件(2012A版)对8家生产企业共30批次小儿感冒颗粒建立指纹图谱,确定共有峰并对其进行相似度评价。测定其中绿原酸、对羟基苯乙酮、木犀草苷、连翘酯苷A、毛蕊花糖苷、4,5-O-二咖啡酰奎宁酸、连翘苷的含量并进行方法学验证。基于指纹图谱共有峰峰面积的测定结果,采用聚类分析、主成分分析、多元统计等方法对不同生产企业的小儿感冒颗粒进行质量评价。结果:建立了小儿感冒颗粒的UPLC指纹图谱,30批样品的相似度均>0.9。确定了17个共有峰,通过与对照品比较指认了其中的8个色谱峰,并对其中7个进行了含量测定。7个成分在各自质量浓度范围内线性关系良好(r≥0.9998),精密度(RSD≤1.1%),重复性(RSD≤1.5%)均良好,供试品在24 h内稳定,平均加样回收率为97.9%~102.6%,RSD为0.71%~1.4%。30批次样品中绿原酸、对羟基苯乙酮、木犀草苷、连翘酯苷A、毛蕊花糖苷、4,5-O-二咖啡酰奎宁酸、连翘苷的含量分别为77.22~160.75、11.04~163.47、26.73~214.65、128.30~1291.02、17.52~120.02、26.51~124.71、57.76~401.96μg·g^(-1)。当欧式距离为5时,8家生产企业可以聚为三类;主成分1~4是影响小儿感冒颗粒生产企业间差异性的主要因子。多元统计分析表明同一生产企业生产的样品,有的生产工艺和原药材的质量控制较为稳定,批间差异性较小;而有的批间差异性较大。其中峰9、1、3、15(连翘苷)、6和11(毛蕊花糖苷)这6个色谱峰所代表的化学成分是引起样品差异性的主要因素。结论:UPLC指纹图谱结合多指标成分�Objective:To establish a quality evaluation method of Xiao'er Ganmao granules based on UPLC fingerprint,multi-component determination,and chemometric analysis.Methods:The Waters Acquity UPLC BEH C_(18)column(100 mm×2.1 mm,1.7μm)was adopted,the mobile phase was acetonitrile-0.2%phosphoric acid aqueous solution with gradient elution.The flow rate was 0.3 mL·min^(-1),the column temperature was 30℃,and multi wavelength switching detection was used.The TCM chromatographic fingerprint similarity evaluation software(2012A version)was used to establish fingerprints of 30 batches of Xiao'er Ganmao granules from 8 manufacturers,the common peaks were identified and the similarity were calculated.The content determination methods for chlorogenic acid,4'-hydroxyacetophenone,luteolin-7-O-glucoside,forsythoside A,verbascoside,3,5-O-dicaffeoylquinic acid,4,5-O-dicaffeoylquinic acid and forsythin were validated.Based on the determination results of the common peak area of the fingerprint,the quality of Xiao'er Ganmao granules from different manufacturers were evaluated by cluster analysis,principal component analysis and multivariate statistics.Results:The UPLC fingerprint of Xiao'er Ganmao granules was established,and the similarities of 30 batches of samples were all above 0.9.Seventeen common peaks were identified,eight of which were identified by comparison with the reference substance,and the contents of seven common peaks were determined.The seven components had good linearity(r≥0.9998),precision(RSD≤1.1%)and repeatability(RSD≤1.5%)within their respective concentration ranges.The samples were stable within 24 h,and the average recoveries were 97.9%-102.6%,with the RSDs of 0.71%-1.4%.The contents of chlorogenic acid,4'-hydroxyacetophenone,luteolin-7-O-glucoside,forsythoside A,verbascoside,3,5-O-dicaffeoylquinic acid,4,5-O-dicaffeoylquinic acid and forsythin in 30 batches of samples were 77.22-160.75,11.04-163.47,26.73-214.65,128.30-1291.02,17.52-120.02,26.51-124.71 and 57.76-401.96μg·g^(-1)respectively.The eight

关 键 词：小儿感冒颗粒 超高效液相色谱 指纹图谱 含量测定 化学计量学 质量评价 

分 类 号：R917[医药卫生—药物分析学]