Niu Chao, Sun Jia, Zheng Yongquan, Wang Linlin, Zhang Jian, Chen Ruijie, Ye Weijian
The Second Affiliated Hospital and Yuying Children's Hospital of Wenzhou Medical University, Wenzhou, 325027, China.
School of Pharmaceutical Science, Wenzhou Medical University, Wenzhou, 325000, China.
J Pharm Biomed Anal. 2020 May 30;184:113210. doi: 10.1016/j.jpba.2020.113210. Epub 2020 Feb 26.
Isosinensetin is a polymethoxyflavone existing in various kinds of citrus. It has exhibited significant anti-proliferative activity and herb-drug interaction. To date, a specific determination method to quantify isosinensetin concentration in biological matrix has not been developed. In the present study, a highly specific, simple and sensitive ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) approach was developed and validated for quantification of isosinensetin in rat plasma with subsequent application to a pharmacokinetic study. Isosinensetin and lysionotin (internal standard, IS) were extracted from rat plasma by a single step protein precipitation using acetonitrile as precipitation agent. The chromatographic separation was conducted using an Agilent C18 column with a gradient elution system (0.1 % formic acid aqueous solution and acetonitrile) within 3.5 min. An electrospray ionization (ESI) source operating in positive mode and multiple reaction monitoring (MRM) were used to monitor the transitions of m/z 373.1 → 343.1 for isosinensetin and m/z 345.1 → 315.1 for IS. The developed method was linear within the range of 1-1000 ng/mL and fully validated according to FDA guidelines. The accuracy values reported as relative errors were between 2.0 and 10.0 % for three quality control levels (2, 400 and 800 ng/mL) and lower limit of quantification (LLOQ). The precisions were ≤11.1 % for quality controls and ≤18.1 % for LLOQ. The recoveries and matrix effects of isosinensetin were in the range of 83.4-87.7 % and 105.6-108.8 %, respectively. Other parameters such as selectivity, carryover effect, dilution integrity and stability were also validated and met the acceptance criteria. The method was applied to a pharmacokinetic study in rats following oral and intravenous administration of isosinensetin. Isosinensetin was rapidly absorbed with a poor bioavailability of 2.19 % and quickly eliminated with mean half-life of 1.40 h and 1.76 h for oral and intravenous route, respectively.
异橙皮素是一种存在于多种柑橘类水果中的多甲氧基黄酮。它已表现出显著的抗增殖活性和药草-药物相互作用。迄今为止，尚未开发出一种用于定量生物基质中异橙皮素浓度的特定测定方法。在本研究中，开发并验证了一种高特异性、简单且灵敏的超高效液相色谱-串联质谱（UHPLC-MS/MS）方法，用于定量大鼠血浆中的异橙皮素，并随后应用于药代动力学研究。使用乙腈作为沉淀剂，通过单步蛋白沉淀从大鼠血浆中提取异橙皮素和裂环马钱子苷（内标，IS）。使用安捷伦C18柱在3.5分钟内采用梯度洗脱系统（0.1%甲酸水溶液和乙腈）进行色谱分离。采用正模式运行的电喷雾电离（ESI）源和多反应监测（MRM）来监测异橙皮素的m/z 373.1 → 343.1转变以及内标的m/z 345.1 → 315.1转变。所开发的方法在1-1000 ng/mL范围内呈线性，并根据FDA指南进行了全面验证。对于三个质量控制水平（2、400和800 ng/mL）以及定量下限（LLOQ），以相对误差报告的准确度值在2.0%至10.0%之间。质量控制的精密度≤11.1%，LLOQ的精密度≤18.1%。异橙皮素的回收率和基质效应分别在83.4-87.7%和105.6-108.8%范围内。其他参数如选择性、残留效应、稀释完整性和稳定性也经过验证并符合验收标准。该方法应用于大鼠口服和静脉注射异橙皮素后的药代动力学研究。异橙皮素吸收迅速，生物利用度较差，为2.19%，消除迅速，口服和静脉途径的平均半衰期分别为1.40小时和1.76小时。
