A rapid LC/MS/MS quantitation assay for naringin and its two metabolites in rats plasma
A rapid LC/MS/MS quantitation assay for naringin and its two metabolites in rats plasma
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DOI：
10.1016/j.jpba.2005.07.031
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发表时间：
2006-02-13
影响因子：
3.4
通讯作者：
Zhao, PY
中科院分区：
文献类型：
--
作者：
Fang, TZ;Wang, YG;Zhao, PY
柚皮苷是一种黄酮类化合物，存在于许多植物和传统中药中。本研究建立了一种高灵敏度、高特异性的电喷雾液相色谱-串联质谱（LC/MS/MS）联用技术，用于柚皮苷及其代谢产物柚皮素和柚皮素葡萄糖醛酸苷的定量分析。以橙皮苷为内标，用乙酸乙酯从大鼠血浆中提取柚皮苷和柚皮素。提取物中的组分在100 mm × 2.0 mm Betabasic 5 μ m C-18 ODS柱上用70%甲醇等度洗脱分离。在多反应监测（MRM）模式下，以m/z 581.3/273.4（柚皮苷）、m/z 273.4/153.1（柚皮素）和m/z 611.5/303.4（橙皮苷）的前体/产物离子对分别对组分进行分析。使用0.1 ml大鼠血浆，在5-1000 ng/ml范围内获得线性校准曲线。在5、50和500 ng/ml（n = 5）下，柚皮苷的日内变异系数（CV）分别为3.1、1.8和2.2%，柚皮素为3.0、3.3和3.1%。在5、50和500 ng/ml时，柚皮苷的日间CV分别为3.4、1.7和4.9%，柚皮素的日间CV分别为4.0、3.0和4.6%（n = 5）。使用基于PEG 400的制剂并经口给予Sprague-Dawley雄性大鼠。用此方法测定血浆药物浓度，用WinNonlin计算机软件进行药代动力学分析。柚皮苷的血药浓度-时间曲线在2 h内迅速上升，24 h后迅速下降，24 h后检测不到。柚皮素和柚皮素葡糖苷酸发生较慢，Tmax分别约为9和7.5小时后。(c)2005 Elsevier B. V.保留所有权利。
Naringin is a flavonoid that exists in many plants and traditional Chinese medicines. In this study, a highly sensitive and specific electrospray ionization (ESI) liquid chromatography-tandem mass spectrometry (LC/MS/MS) method was developed for quantification of naringin and its two metabolites, naringenin and naringenin glucuronide. Naringin and naringenin were extracted from rat plasma with ethyl acetate, using hesperidin as an internal standard. Components in the extract were separated on a 100 mm x 2.0 mm Betabasic 5 mu m C-18 ODS column by isocratic elution with 70% methanol. The components were analyzed in the multiple-reaction-monitoring (MRM) mode in the precursor/product ion pair of m/z 581.3/273.4 for naringin, m/z 273.4/153.1 for naringenin and m/z 611.5/303.4 for hesperidin, respectively. Linear calibration curves were obtained in the range of 5-1000 ng/ml, using 0.1 ml rat plasma. The within-day coefficients of variation (CVs) were 3.1, 1.8 and 2.2% for naringin, 3.0, 3.3, 3.1% for naringenin at 5, 50 and 500 ng/ml (n = 5). The between-day CVs were 3.4, 1.7 and 4.9% for naringin and 4.0, 3.0, 4.6% for naringenin (n = 5) at 5, 50 and 500 ng/ml respectively. A formulation based on PEG400 was used and orally administered to Sprague-Dawley male rats. Plasma drug concentrations were measured by this method and the pharmacokinetics was analyzed by WinNonlin computer software. Plasma concentration-time profiles of naringin were found to increase quickly and decline rapidly within 2 h and could not be detected after 24 h. Naringenin and naringenin glucuronide occurred slower and the T-max were about 9 and 7.5 h later, respectively. (c) 2005 Elsevier B.V. All rights reserved.