目的:建立甘地胶囊中咖啡酸、阿魏酸、芦丁、野黄芩苷、黄芩苷、黄芪甲苷、汉黄芩素7个成分的UHPLC—MS/MS测定方法。方法:采用ZORBAXSB—C18(2.1mm×50mm,1.8μm)色谱柱,以乙腈(A)一水(B)为流动相,梯度洗脱(0—4min,5%A-20%A;4~4.5min,20%A→40%A;4.5—8min,40%A→43%A;8—8.5min,43%A→90%A;8.5~9min,90%A→5%A),流速0.4mL·min^-1,柱温35℃;采用电喷雾离子源(ESI),多反应离子监测(MRM),结合正负离子扫描切换,其中咖啡酸、阿魏酸、野黄芩苷采用负离子模式检测,芦丁、黄芩苷、黄芪甲苷、汉黄芩素采用正离子模式检测。结果:咖啡酸、阿魏酸、芦丁、野黄芩苷、黄芩苷、黄芪甲苷、汉黄芩素的定量下限分别为0.22、0.35、0.39、0.48、0.31、1.31、0.11ng·mL^-1,检测下限分别为0.066、0.11、0.12、0.14、0.092、0.39、0.033ng·mL^-1;在相应的线性范围内相关系数〉0.99;日内和日间精密度(RSD)均小于5%,平均回收率均为80%~120%。结论:经方法学验证,本方法简单、快速、灵敏、准确,可用于甘地胶囊中咖啡酸、阿魏酸、芦丁、野黄芩苷、黄芩苷、黄芪甲苷、汉黄芩素7个成分的含量测定。通过对18个批次甘地胶囊进行测定,结果显示各个批次含量有较明显差别,同时随着时间推移,含量略有下降趋势,尤其以芦丁和黄芩苷的变化趋势较明显,可为该药的质量控制提供依据。
Objective: To establish a UHPLC - MS/MS method for simultaneous determination of caffeic acid,ferulic acid, rutin, scutellarin, baicalin, astragaloside IV, wogonin in Gandi capsules. Methods: The chromatographic separation was on ZORBAX SB -C 18 column (2. 1 mm× 50 mm, 1.8 μm) with a mobile phase consisting of acetonitrile (A) -water (B) at a flow rate of 0.4 mL· min^-1(0 -4 min,5%A→20%A;4 -4. 5 min,20% A→40% A;4. 5 -8 min, 40% A→3 % A ; 8 - 8. 5 min,43 % A→90% A ; 8. 5 - 9 min,90% A→5% A), and the column temperature was 35 ℃, using electrospray ionization source (ESI), and muhiple reaction monitoring (MRM), combined with positive and negative scanning switch. Caffeic acid, ferulic acid, and scutellarin were detected using negative ion mode,while rutin, baicalin,as- tragaloside IV,and wogonin were detected using positive ion mode. Results: The quantification limits of caffeic acid ,feru- lie acid, rutin, seutellarin, baicalin, astragaloside IV, and wogonin were 0. 22,0. 35, 0. 39,0. 48,0. 31,1.31,0. 11 ng · mL^-1 ,and the detection limits were 0. 066,0. 11,0. 12,0. 14,0. 092,0. 39,0. 033 ng· mL^-1 ,respectively. Within the linear range,the correlation coefficient was 〉0. 99. Intraday and inter- day precisions (RSDs) were less than 5%. The aver- age recoveries of the seven components were in the range of 80% - 120%. Conclusion: By method validation ,this method is proved to be simple,rapid, sensitive and accurate. It can be used to determine eaffeic acid ,ferulic acid, rutin, scutel-larin ,baicalin, astragaloside IV, and wogonin in Gandi capsules. The determination results of 18 batches of Gandi capsules showed that the content of each batch had significant difference, and the content had a slight decreasing trend over time, especially for rutin and baicalin. These results were helpful for the quality control of drug.