CAJ | 학술논문

目的:建立黄芪甲苷(astragaloside, AGS-Ⅳ)在完整大鼠体内血药浓度的测定方法及初步的药动学评价.方法:运用液质联用仪(HPLC-MS)测定大鼠血浆AGS-Ⅳ浓度.6只雄性SD大鼠,AGS-Ⅳ溶液静脉给药(2.0 mg/kg),单只大鼠连续取血法,取血点分别为 0.025、 0.05、 0.1、 0.25、 0.5、 1、 2、 4、 6、 10、 14和 24 h.固相萃取小柱提取血浆样品,地高辛为内标(I.S.) ,LC-ESI-MS测定血药浓度.AGS-Ⅳ的m/z为 807.5,内标地高辛的m/z为 803.5.结果:AGS-Ⅳ的标准曲线线性范围为1～1 000 ng/mL (r=0.9992),日内和日间精密度分别小于6%和8%,血浆样品AGS-Ⅳ的回收率为 92.8%～98.4%,内标的回收率为 80.0%～90.9%,最低检测限为 0.5 ng/mL.CAPP软件拟合,AGS-Ⅳ的消除符合二室模型,药动学参数t1/2β(h),CL(L·kg-1·h),Vc(L/kg),AUC0-∞(μg·mL-1·h)分别为:3.46±0.52, 0.47±0.02, 0.76±0.16 和4274±186.结论:连续间断取血法结合HPLC-MS技术,适用于测定AGS-Ⅳ在小动物的血药浓度和药动学评价.
목적:건립황기갑감(astragaloside, AGS-Ⅳ)재완정대서체내혈약농도적측정방법급초보적약동학평개.방법:운용액질련용의(HPLC-MS)측정대서혈장AGS-Ⅳ농도.6지웅성SD대서,AGS-Ⅳ용액정맥급약(2.0 mg/kg),단지대서련속취혈법,취혈점분별위 0.025、 0.05、 0.1、 0.25、 0.5、 1、 2、 4、 6、 10、 14화 24 h.고상췌취소주제취혈장양품,지고신위내표(I.S.) ,LC-ESI-MS측정혈약농도.AGS-Ⅳ적m/z위 807.5,내표지고신적m/z위 803.5.결과:AGS-Ⅳ적표준곡선선성범위위1～1 000 ng/mL (r=0.9992),일내화일간정밀도분별소우6%화8%,혈장양품AGS-Ⅳ적회수솔위 92.8%～98.4%,내표적회수솔위 80.0%～90.9%,최저검측한위 0.5 ng/mL.CAPP연건의합,AGS-Ⅳ적소제부합이실모형,약동학삼수t1/2β(h),CL(L·kg-1·h),Vc(L/kg),AUC0-∞(μg·mL-1·h)분별위:3.46±0.52, 0.47±0.02, 0.76±0.16 화4274±186.결론:련속간단취혈법결합HPLC-MS기술,괄용우측정AGS-Ⅳ재소동물적혈약농도화약동학평개.
AIM: To establish a sensitive method for quantitative determination of astragaloside Ⅳ (AGS-Ⅳ) in plasma and a preliminary evaluation of its pharmacokinetics parameters in intact rats. METHODS: A liquid chromatography-electrospray ionization-mass spectrometry (LC-ESI-MS) was applied for determining AGS-Ⅳ in plasma by using digoxin as the internal standard (I.S.). Six rats were given AGS-Ⅳ 2.0 mg/kg by intravenous infusion for 5 min. Blood samples were drawn intermittently with each intact rat from left femoral artery at 0.025, 0.05, 0.1, 0.25, 0.5, 1, 2, 4, 6, 10, 14 and 24 h after medication. The samples were prepared by solid phase extraction and analyzed through a triple quadrupole mass spectrometer equipped with an electrospary probe. The samples were monitored in selected ion recording (SIR) mode of positive ions by using target ions at m/z 807.5 for AS- Ⅳand at m/z 803.5 for I.S. RESULTS: Calibration curves were linear over the ranges 1-1 000 ng/mL for AGS-Ⅳ (r=0.9992). The intra-and inter-day assay variability values were less than 6% and 8%, respectively. Extraction recoveries from plasma were 92.8%-98.4% for AGS-Ⅳ and 80.0%-90.9% for digoxin, respectively. The lower limit of quantitation (LLOQ) for AGS-Ⅳ was 0.5 ng/mL. The concentration-time curves of AGS-Ⅳ for each rat were fitted to an open two-compartment model by CAPP program. The pharmacokinetics parameters of AGS-Ⅳ were as following: the elimination half-life (t1/2β), clearance rate (CL), distribution volume at steady state (Vss), and AUC0-∞ were (3.46±0.52) h, (0.47±0.02) L/h, (0.76±0.16) L/kg and (4.27±0.19) μg·mL-1·h, respectively. CONCLUSION: These results show that this method is satisfied for the measurements of pharmacokinetics study for AGS-Ⅳ.