Department of Drug Metabolism, Institute of Materia Medica, Chinese Academy of Medical Sciences and Peking Union Medical College, 1 Xian Nong Tan Street, Beijing 100050, PR China; Beijing Key Laboratory of Non-Clinical Drug Metabolism and PK/PD Study, Institute of Materia Medica, Chinese Academy of Medical Sciences and Peking Union Medical College, 1 Xian Nong Tan Street, Beijing 100050, PR China; Beijing Key Laboratory of Active Substances Discovery and Drug Ability Evaluation, Institute of Materia Medica, Chinese Academy of Medical Sciences and Peking Union Medical College, 1 Xian Nong Tan Street, Beijing 100050, PR China; State Key Laboratory of Bioactive Substance and Function of Natural Medicines, Institute of Materia Medica, Chinese Academy of Medical Sciences and Peking Union Medical College, 1 Xian Nong Tan Street, Beijing 100050, PR China.
State Key Laboratory of Bioactive Substance and Function of Natural Medicines, Institute of Materia Medica, Chinese Academy of Medical Sciences and Peking Union Medical College, 1 Xian Nong Tan Street, Beijing 100050, PR China.
J Pharm Biomed Anal. 2021 Jan 30;193:113720. doi: 10.1016/j.jpba.2020.113720. Epub 2020 Nov 2.
N6-[(S)-1- (phenyl)-propyl]-adenine riboside (YZG-331) is being developed as a novel sedative and hypnotic agent. The hydroxylated metabolites of YZG-331 have the same mass transition ion pair, making their determination in blood challenging. In this study, a rapid and sensitive liquid chromatography-tandem mass spectrometry method was developed for the simultaneous determination of YZG-331 and its metabolites M1 (hydrolysis), M2 and M4 (hydrolysis and hydroxylation), M3, M5 and M6 (hydroxylation) in monkey blood. Propranolol was used as the internal standard (IS). Blood samples were prepared using a simple protein precipitation with acetonitrile. The chromatographic separation was performed on an Eclipse Plus C18 column (2.1 × 50 mm, 3.5 μm) at a flow rate of 0.3 mL/min with a gradient mobile phase of methanol/water containing 0.5 % formic acid (v/v). Detection was carried out on a triple quadrupole mass spectrometer in positive ion multiple reaction monitoring mode. The optimized mass transition ion pairs for quantitation were 386→254 for YZG-331, 254→136 for M1, 270→136 for M2 and M4, 402→136 for M3, M5 and M6 and 260→183 for IS. Acceptable linearity was obtained for the analytes over the range of 15-2000 ng/mL for YZG-331, 3-400 ng/mL for M1-M6. The lower limits of the quantification were 15 ng/mL for YZG-331, 3 ng/mL for M1-M6. The intra- and inter-day precisions wre within 10.5 % for all analytes, while the accuracy ranged from -8.3 %-8.8 %. There was no obvious matrix effect and the recoveries of the analytes were 90.6 %-118.2 %. The analytes were proved to be stable during all sample storage, preparation and analytic procedures. The sensitive and rapid LC-MS/MS method for YZG-331 in monkey blood has been applied to pharmacokinetic studies of YZG-331 in monkeys. The oral bioavailability of YZG-331 in monkeys is 74.1 %.
N6-[(S)-1- (苯基)-丙基]-腺嘌呤核苷（YZG-331）正在被开发为一种新型的镇静和催眠药物。YZG-331 的羟基化代谢物具有相同的质量转换离子对，这使得它们在血液中的测定具有挑战性。在这项研究中，开发了一种快速灵敏的液相色谱-串联质谱法，用于同时测定猴血中的 YZG-331 及其代谢物 M1（水解）、M2 和 M4（水解和羟基化）、M3、M5 和 M6（羟基化）。普萘洛尔被用作内标（IS）。血液样品用乙腈简单沉淀蛋白后制备。色谱分离在 Eclipse Plus C18 柱（2.1×50mm，3.5μm）上进行，流速为 0.3mL/min，梯度流动相为甲醇/水，含有 0.5%（v/v）甲酸。检测采用三重四极杆质谱仪在正离子多重反应监测模式下进行。定量的优化质量转换离子对分别为 386→254 用于 YZG-331、254→136 用于 M1、270→136 用于 M2 和 M4、402→136 用于 M3、M5 和 M6、260→183 用于 IS。YZG-331 在 15-2000ng/mL 范围内，M1-M6 在 3-400ng/mL 范围内，分析物均获得可接受的线性。YZG-331 的定量下限为 15ng/mL，M1-M6 的定量下限为 3ng/mL。所有分析物的日内和日间精密度均在 10.5%以内，准确度范围为-8.3%-8.8%。无明显基质效应，分析物回收率为 90.6%-118.2%。在所有样品储存、制备和分析过程中，分析物均稳定。该灵敏快速的 LC-MS/MS 法已应用于猴体内 YZG-331 的药代动力学研究。YZG-331 在猴体内的口服生物利用度为 74.1%。
