HPLC-MS(TOF)法测定家兔血浆中黄芪甲苷的浓度

Determination of astragaloside Ⅳ in rabbit plasma by HPLC-MS

目的建立测定家兔血浆中黄芪甲苷的HPLCMS(TOF)法。方法含药血浆用C18固相萃取小柱纯化样品,用LC/MS(TOF)联用技术,以电喷雾(ESI)作为接口技术,选择黄芪甲苷的离子([M+Na]+807)和内标人参皂甘Rg1的离子([M+Na]+823)作为测定离子,测定血浆中黄芪甲苷的浓度。结果黄芪甲苷的回归方程AsAi=1.2668×103C+0.096557,r=0.999;线性范围为10～5000ng·mL-1,定量限为5ng·mL-1,回收率大于90%。结论该测定方法灵敏度高、专属性好、快速,可满足黄芪甲苷药代动力学研究要求。

AIM: To establish a sensitive and specific liquid chromatobraphy-mass spectrometry(time-of-flight)[LC-MS(TOF)] for the determination of astragaloside Ⅳ in rabbit plasma. METHODS: The HPLC-MS utilizing solid phase extraction was established to determine the concentration of astragaloside IV and ginsenoside R_~g1 , was used as internal standard. The analysis was carried on Agilent Hypersiol ODS(5 μm, 4.6 mm×250 mm) column with a mobile phase of methanol-water (80∶20, v/v).Detection was performed on a time-of-flight mass spectrometry equipped with an ESI internal and operated in positive-ionization mode. Astragaloside Ⅳ quantitation was realized by computing the peak area ratio (astragaloside Ⅳ-ginsenoside R_~g1 )(astragaloside Ⅳ m/z807[M+Na]+ and ginsenoside R_~g1 m/z823[M+Na]+) and comparing them with calibration curve (r=~0.999 ). RESULTS: The linear calibration curve was obtained in the concentration range of 0.01-5 μg·L^-1 .The detection limit of astragaloside Ⅳ was 0.01 μg·L^-1 .The average recovery was more than 98%.The intra-and inter-run precision was measured to be below 5% of RSD. CONCLUSION: The method is sensitive, simple and rapid ,so, it can meet the need for the pharmacokinetics and bioavailability of astragaloside Ⅳ.