摘要
目的:建立同时测定金藤清痹颗粒中青藤碱、新绿原酸、木兰花碱、隐绿原酸、绿原酸、异绿原酸B、异绿原酸A、异绿原酸C、哈巴俄苷、藁本内酯和甘草酸铵11个成分含量的UHPLC法,并结合主成分分析对制剂进行质量评价.方法:采用UHPLC波长切换法,色谱柱为Waters XSelect ® CSH C18柱(150 mm ×4.6 mm,2.5 µm),以甲醇-乙腈(1∶1)为流动相A,0.2％磷酸水溶液为流动相B,梯度洗脱,流速1.0 mL·min-1,柱温25℃,检测波长218 nm(0～17 min时,检测青藤碱)、326 nm(17～25 min和34～98 min时,检测新绿原酸、隐绿原酸、绿原酸和异绿原酸A、异绿原酸B、异绿原酸C)、263 nm(25～34 min和98～125 min时,检测木兰花碱、哈巴俄苷、藁本内酯、甘草酸铵).利用SPSS27.0软件对20批金藤清痹颗粒中成分含量进行多元统计分析.结果:青藤碱、新绿原酸、木兰花碱、隐绿原酸、绿原酸、异绿原酸B、异绿原酸A、异绿原酸C、哈巴俄苷、藁本内酯和甘草酸铵11个成分的质量浓度分别在 14.36～143.61 µg·mL-1(r=0.999 9)、7.71～77.15 μg·mL-1(r=0.999 8)、9.18～91.83 μg·mL-1(r=0.999 7)、10.71～107.07 μg·mL-1(r=0.999 8)、12.88～128.80 μg·mL-1(r=0.999 8)、5.20～51.95 μg·mL-1(r=0.999 7)、5.18～51.84μg·mL-1(r=0.999 8)、5.40～53.95 μg·mL-1(r=0.999 8)、2.62～26.16 μg·mL-1(r=0.999 9)、6.31～63.06 μg·mL-1(r=0.999 9)和 11.13～111.26 μg·mL-1(r=0.997 6)范围内与峰面积的线性关系良好;平均加样回收率(n=6)分别为98.3％、98.3％、98.5％、98.9％、99.2％、101.0％、98.1％、97.1％、96.8％、98.0％和 98.7％,RSD 均小于 3.0％.20 批金藤清痹颗粒样品中,上述青藤碱等11个成分的含量测定结果(n=6)依次为2.206～2.704、1.071～1.403、2.096～2.487、1.321～1.724、2.241～2.612、0.605～0.749、0.363～0.412、0.835～1.020、0.151～0.191、0.791～1.188和1.008～1.363 mg·g-1.主成分分析结果显示,连续生产的金藤清痹颗粒批次间质量差异较小,且以样品S1、S5和S7的综合质量相对更好.结论:建立的UHPLC多指标成分含量测定方法简便、准确、稳定,结合主成分分析,可全面地评价金藤清痹颗粒的产品质量.
Abstract
Objective:To establish a UHPLC method for simultaneously determining sinomenine,neochlorogenic acid,magnolflorine,cryptochlorogenic acid,chlorogenic acid,isochlorogenic acid B,isochlorogenic acid A,isochlorogenic acid C,harpagoside,ligustilide and ammonium glycyrrhizate in Jinteng Qingbi granules,and evalu-ate the quality of Jinteng Qingbi granules combined with principal component analysis.Methods:UHPLC wave-length switching method was employed in the study.The chromatographic separation was performed on a Waters XSelect ® CSH C18 column(150 mm ×4.6 mm,2.5 μm)using methanol-acetonitrile(1∶1)as mobile phase A and 0.2％phosphoric acid aqueous solution as mobile phase B in a gradient mode at 25 ℃.The flow rate was 1.0 mL·min-1,and the UV detection wavelength was chosen at 218 nm for sinomenine during 0-17 min,326 nm for neochlorogenic acid,cryptochlorogenic acid,chlorogenic acid,isochlorogenic acid B,isochlorogenic acid A and isochlorogenic acid C during 17-25 min and 34-98nm,263 nm for magnolflorine,harpagoside,ligustil-ide and ammonium glycyrrhizate during 25-34 min and 98-125nm,respectively.Furthermore,multiple statisti-cal analysis was conducted on the contents of 11 components in 20 batches of Jinteng Qingbi granules using SPSS27.0 software.Resluts:Satisfactory linearities of sinomenine,neochlorogenic acid,magnolflorine,crypto-chlorogenic acid,chlorogenic acid,isochlorogenic acid B,isochlorogenic acid A,isochlorogenic acid C,harpago-side,ligustilide and ammonium glycyrrhizate were in the ranges of 14.36-143.61 μg·mL-1(r=0.999 9),7.71-77.15 μg·mL-1(r=0.999 8),9.18-91.83 μg·mL-1(r=0.999 7),10.71-107.07 μg·mL-1(r=0.999 8),12.88-128.80 μg·mL-1(r=0.999 8),5.20-51.95 μg·mL-1(r=0.999 7),5.18-51.84 μg·mL-1(r=0.999 8),5.40-53.95 μg·mL-1(r=0.999 8),2.62-26.16 μg·mL-1(r=0.999 9),6.31-63.06 μg·mL-1(r=0.999 9)and 11.13-111.26 μg·mL-1(r=0.997 6),respec-tively.The average recoveries(n=6)were 98.3％,98.3％,98.5％,98.9％,99.2％,101.0％,98.1％,97.1％,96.8％,98.0％and 98.7％,respectively,with RSDs less than 3.0％.The contents ranges of sinome-nine and other 10 components of 20 batches of Jinteng Qingbi granules samples were 2.206-2.704,1.071-1.403,2.096-2.487,1.321-1.724,2.241-2.612,0.605-0.749,0.363-0.412,0.835-1.020,0.151-0.191,0.791-1.188 and 1.008-1.363 mg·g-1,respectively.The results of principal component a-nalysis showed that the quality differences between batches of continuously produced Jinteng Qingbi granules were relatively smaller,and the comprehensive quality of samples S1,S5 and S7 was relatively better.Conclusion:The established UHPLC method of multi-index component determination is simple,accurate and stable.Combined with principal component analysis,it can be used for quality evaluation of Jinteng Qingbi granules comprehensively.
基金项目
山东省重点研发计划(重大科技创新工程)(2021CXGC010508)
山东省新旧动能转换重大产业攻关项目(鲁动能办[2021]23号)
NETL
NSTL
万方数据