Liao Sha, Ren Qiuxia, Yang Cuiping, Zhang Tianhong, Li Jinglai, Wang Xiaoying, Qu Xinyan, Zhang Xiaojuan, Zhou Zhe, Zhang Zhenqing, Wang Shengqi
Beijing Institute of Pharmacology and Toxicology , 27 Taiping Road, Haidian District, Beijing, 100850 P. R. China.
J Agric Food Chem. 2015 Feb 25;63(7):1957-66. doi: 10.1021/jf5019615. Epub 2015 Feb 16.
Amentoflavone (AMF) is a biflavone found in many herbal dietary supplements. To investigate the pharmacokinetic profile of AMF in rats, a sensitive, simple, and accurate liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed and used to monitor AMF and its conjugated metabolites in plasma. AMF was administered to rats by oral gavage (po), or by intravenous (iv) or intraperitoneal (ip) injection. Plasma samples (with apiolin as an internal standard) were liquid/liquid extracted after hydrolysis with β-glucuronidase/sulfatase in vitro. Following chromatographic separation on a C18 column with a methanol:water:formic acid (70:30:0.1, v/v/v) mobile phase, AMF and internal standard were determined by electrospray ionization in negative ion mode and their precursor-product ion pairs (m/z 537.1 → 374.9 and m/z 269.2 → 224.9, respectively) were used for measurement. This bioanalytical method was fully validated and showed good linearity (r(2) > 0.99), wide dynamic range (0.93-930 nmol/L), and favorable accuracy and precision. After iv or ip AMF (10 mg/kg) injection, 73.2% ± 6.29% and 70.2% ± 5.18% of the total AMF detected in plasma was present as conjugated metabolites. Furthermore, AMF and AMF conjugates showed similar time courses with no significant differences in the time to reach the maximum plasma concentration (tmax) and terminal half-life (t1/2) (p > 0.05). Following po AMF administration (300 mg/kg), 90.7% ± 8.3% of the total AMF was circulating as conjugated metabolites. When compared with iv administration (with dose correction), the bioavailability of po AMF was very low (0.04% ± 0.01% for free AMF; 0.16% ± 0.04% for conjugated AMF). Collectively, these data provided a preliminary pharmacokinetic profile for AMF that should inform further evaluations of its biological efficacy and preclinical development.
穗花杉双黄酮（AMF）是一种存在于多种草本膳食补充剂中的双黄酮。为了研究AMF在大鼠体内的药代动力学特征，开发了一种灵敏、简便且准确的液相色谱 - 串联质谱（LC-MS/MS）方法，并用于监测血浆中的AMF及其共轭代谢物。通过口服灌胃（po）、静脉注射（iv）或腹腔注射（ip）的方式给大鼠施用AMF。血浆样品（以芹菜素为内标）在体外经β - 葡萄糖醛酸酶/硫酸酯酶水解后进行液 - 液萃取。在以甲醇:水:甲酸（70:30:0.1，v/v/v）为流动相的C18柱上进行色谱分离后，采用负离子模式下的电喷雾电离测定AMF和内标，其前体 - 产物离子对（分别为m/z 537.1 → 374.9和m/z 269.2 → 224.9）用于测量。这种生物分析方法经过了全面验证，显示出良好的线性（r(2) > 0.99）、宽动态范围（0.93 - 930 nmol/L）以及良好的准确度和精密度。静脉注射或腹腔注射AMF（10 mg/kg）后，血浆中检测到的总AMF的73.2% ± 6.29%和70.2% ± 5.18%以共轭代谢物的形式存在。此外，AMF及其共轭物显示出相似的时间进程，在达到最大血浆浓度（tmax）和末端半衰期（t1/2）的时间上无显著差异（p > 0.05）。口服AMF（300 mg/kg）后，总AMF的90.7% ± 8.3%以共轭代谢物的形式循环。与静脉注射（进行剂量校正后）相比，口服AMF的生物利用度非常低（游离AMF为0.04% ± 0.01%；共轭AMF为0.16% ± 0.04%）。总体而言，这些数据提供了AMF的初步药代动力学特征，应为其生物学功效的进一步评估和临床前开发提供参考。
