LC-MS/MS同时测定大鼠血浆中黄芪成分芒柄花素、毛蕊异黄酮和异鼠李素的浓度及其药动学研究  

Simultaneous Determination of Formononetin,Calycosin and Isorhamnetin from Astragalus mongholicus in Rat Plasma by LC-MS /MS and Application to Pharmacokinetic Study

作　　者：林青[1] 李媛[1] 谭晓梅[1] 姚向超[2] 

机构地区：[1]中山大学附属第一医院黄埔院区,广东广州510700 [2]中山大学中山眼科中心,广东广州510060

出　　处：《中药材》2013年第4期589-593,共5页Journal of Chinese Medicinal Materials

摘　　要：目的:建立大鼠血浆中芒柄花素、毛蕊异黄酮和异鼠李素浓度的测定方法,并研究其药动学参数。方法:取大鼠10只灌胃给予10 g/kg黄芪,检测给药前和给药后24 h内芒柄花素、毛蕊异黄酮和异鼠李素的血浆浓度,并计算其药动学参数;采用液相色谱-串联质谱法,以芦丁为内标,色谱柱为Alltima C18,流动相:甲醇-水溶液,梯度洗脱,流速为0.3 mL/min,柱温为40℃,电喷雾负离子源,扫描方式为多反应离子监测。芒柄花素、毛蕊异黄酮、异鼠李素和芦丁的选择检测离子质荷比(m/z)分别为m/z 267.0→251.9,m/z 283.1→268.2,m/z 315.4→300.1和m/z 609.4→300.1。结果:芒柄花素、毛蕊异黄酮和异鼠李素检测浓度的线性范围分别为5～1 000(r=0.9996)、3.91～500(r=0.9989)、0.5～100(r=0.9992)ng/mL,最低检测限分别为0.625、0.5和0.1 ng/mL,药动学参数分别为:t1/2β(10.43±2.94)、(6.91±1.33)和(5.07±1.21)h,Cmax(398.5±103.7)、(138.7±32.8)和(58.3±14.5)ng/mL,AUC0→12h(1238.8±311.3)、(669.5±159.7)和(274.1±83.9)ng.h/mL。结论:本方法专属性强、灵敏度高、准确性好,可用于芒柄花素、毛蕊异黄酮和异鼠李素的血药浓度测定,大鼠灌胃给10 g/kg黄芪后,芒柄花素、毛蕊异黄酮和异鼠李素在大鼠体内的药动学符合二室模型特征。Objective：To establish a method to determine the concentration of formononetin, calycosin and isorhamnetin from Astragalus mongholicus in rats＇ plasma using LC-MS/MS and calculate their pharmacokinetic parameters. Methods：The contents of formononetin, calycosin and isorhamnetin in plasma were detected before and 24 h after 10 rats were treated with 10 g/kg Astragalus mongholicus. Rutin was used as internal standard. Agilent 1 200 HPLC system with Alltima C18 （150 mm × 2. 1 mm,5 μm）was used. Mobile phase was methanol-water solution with gradient elute at a flow rate of 0. 3 mL/min. The column temperature was 40℃. The LC-MS/ MS system was operated using an electrospray ionization probe in negative ion mode; Scan mode： multiple reaction ion monitoring （ MRM ） mode. The ion of monitor： m/z 267. 0-251.9 for formononetin, m/z 283.1-268. 2 for calycosin, m/z 315.4-300. 1 for isorhamnetin and m/z 609.4-300. 1 for rutin （ internal standard）, respectively. Results： The linear range of formononetin, calycosin and isorhamnetin was 5 - 1 000（ r =0. 9996） ,3.91 - 500 （ r =0. 9989） and 0. 5 - 100 ng/mL（ r =0. 9992）, respectively. The lowest limit of quantification （LLOQ） of formononetin, calycosin and isorhamnetin was 0. 625,0. 5 and 0. 1 ng/mL, respectively. The pharmacokinetic parameter, t1/2β, of formononetin, calycosin and isorhamnetin was （ 10. 43 ± 2. 94 ）, （6. 91 ± 1.33 ） and （ 5.07 ＋ 1.21 ） h, respectively. The Crux of formononetin, calycosin and isorhamnetin was （398. 5 ±103.7 ）, （ 138. 7± 32. 8 ） and （58.3 ± 14. 5 ）ng/mL, respectively. The AUC1-12h of formononetin,calycosin and isorhamnetin was（ 1238. 8 ±311.3）, （669. 5 ± 159.7）and（274. 1±83.9）ng ·h/mL, respectively. Conclusion：A sensitive,accuracy and suitable LC-MS/MS method for determination of formononetin,calycosin and isorhamnetin is developed and successfully applied to the pharmacokinetic study of 10 g/kg Astragalus mongholicus after oral administration in rats.

关 键 词：黄芪 芒柄花素 毛蕊异黄酮 异鼠李素 液相色谱-串联质谱法 药动学参数 

分 类 号：R285.5[医药卫生—中药学]