液质联用研究骨碎补提取物给予大鼠后柚皮苷及其代谢物柚皮素的药动学行为(英文)

Pharmacokinetics of Naringin and its Metabolite Naringenin in Rats after Oral Administration of Rhizoma Drynariae Extract Assayed by UPLC-MS/MS

目的:建立超高效液相色谱-串联质谱法(UPLC-MS/MS)研究柚皮苷及其代谢物柚皮素在大鼠体内的药动学行为。方法:骨碎补药材中加入50%乙醇加热回流1h,提取物用50% PEG400制成混悬液灌胃给予大鼠。血浆样品处理采用液液萃取法,以乙酸乙酯为提取试剂,对乙酰氨基酚为内标。色谱条件采用ACQUITY UPLCTM BEHC18色谱柱,流动相为乙腈-0.4%冰醋酸(80:20,V/V),流速为0.25mL.min-1,柱温为25℃;质谱条件采用ESI源,负离子检测方式,多反应监测(MRM)。用于定量分析的离子反应分别为m/z578.4→270.7(柚皮苷)、m/z270.7→150.7(柚皮素)和m/z149.9→106.6(内标)。结果:柚皮苷和柚皮素的Cmax分别为(2.56±0.77)×103ng.mL-1和222±45ng.mL-1,Tmax分别为0.67±0.20h和8.0±1.3h,AUC0～t分别为(3.23±0.54)×103ng.mL-1.h和(1.57±0.37)×103ng.mL-1.h,AUC0～∞分别为(3.29±0.54)×103ng.mL-1.h和(1.79±0.43)×103ng.mL-1.h。柚皮苷的t1/2为4.1±0.8h,且在其药时曲线8h处出现另一微小的峰。结论:所建立的UPLC-MS/MS方法符合生物样品分析要求,药动学研究结果为骨碎补的临床应用提供了参考。

AIM： To investigate the pharmacokinetics of naringin and naringenin in rats after oral administration of Rhizoma Drynariae extract. METHODS： The extract of Rhizoma Drynariae were refluxed with 50% ethanol for 1 h and dissolved with 50% PEG 400, and then administered to the rats. Naringin and naringenin were extracted from rat plasma by means of liquid-liquid extrac- tion （LLE） with ethyl acetate. Paracetamol was used as the internal standard （IS）. The analysis was carded out on an ACQUITY UPLCTM BEH C18 column with acetonitrile-0.4% acetic acid （80： 20, V/V） as mobile phase at a flow rate of 0.25 mL.min-l. The detection was carried out via negative electrospray ionization （ESI） mass spectrometry with multiple-reaction monitoring （MRM）. The quantification was performed using the transitions of m/z 578.4→270.7 for naringin, m/z 270.7→150.7 for naringenin and m/z 149.9→106.6 for paracetamol （IS）, respectively. RESULTS： After oral administration of Rhizoma Drynariae extract to rats, the mean Cmax of naringin and naringenin was （2.56 ± 0.77） × 103 ng·mL-1 and 222 ± 45 ng-mL-1, the Tmax was 0.67 ± 0.20 h and 8.0 ± 1.3 h, the AUC0-t was calculated to be （3.23 ± 0.54）× 103 ng·mL-1.h and （1.57 ± 0.37） × 103 ng-mL-1.h, the AUC0-∞was calculated to be （3.29 ± 0.54） × 103 ng.mL-1-h and （1.79 ± 0.43） × 103 ng-mL-1.h. The t1/2 was 4.1 ± 0.8 h for naringin, and another small peak could be seen at 8h in the plasma concentration-time profile of naringin. CONCLUSION： The developed UPLC-MS/MS method was conformed to the criteria for the analysis of biological samples according to the guidance of FDA. The results of pharmacokinetic study on naringin and its active metabolite naringenin would be a suitable reference in clinical application for Rhizoma Drynariae.