Beijing City Key Laboratory of Active Substances Discovery and Drugability Evaluation, State Key Laboratory of Bioactive Substance and Function of Natural Medicines, Department of Drug Metabolism of Institute of Materia Medica, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100050, China.
Beijing City Key Laboratory of Active Substances Discovery and Drugability Evaluation, State Key Laboratory of Bioactive Substance and Function of Natural Medicines, Department of Drug Metabolism of Institute of Materia Medica, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100050, China.
J Chromatogr B Analyt Technol Biomed Life Sci. 2014 Jan 15;945-946:193-8. doi: 10.1016/j.jchromb.2013.11.005. Epub 2013 Nov 26.
SYL-1119 is a sphingosine-1-phosphate receptor 1 modulator for the treatment of autoimmune disease with better selectivity, while SYL-1119-P is its active phosphate. A sensitive and specific liquid chromatography-tandem mass spectrometry method was developed and validated for the simultaneous determination of SYL-1119 and SYL-1119-P in rat plasma. SYL-1110, an analogue of SYL-1119, was used as the internal standard. Plasma samples were prepared by protein precipitation using acetonitrile. The analytes and internal standard were separated on a Zorbax SB-C18 column (3.5μm, 100mm×2.1mm) with a gradient mobile phase consisting of methanol and water containing 0.1% formic acid at a flow rate of 0.2ml/min with an operating temperature of 20°C. The detection was performed on a triple quadrupole tandem mass spectrometer with positive electrospray ionization in multiple reaction monitoring mode of the transitions at m/z 364→259 for SYL-1119, m/z 444→259 for SYL-1119-P, and m/z 378→273 for the IS. Calibration curves were linear in the range of 0.2-50ng/ml for SYL-1119 and 10-1000ng/ml for SYL-1119-P. The lower limit of quantification (LLOQ) was 0.2ng/ml for SYL-1119 and 10ng/ml for SYL-1119-P. The intra- and inter-day precisions were 5.4-12.8% for two analytes with accuracies within ±10%. The recoveries for two compounds were 91.3-104.5%. The analytes were proved to be stable during all sample storage, preparation, and analytic procedures. The method was successfully applied to the pharmacokinetic study of SYL-1119 and SYL-1119-P in rats after oral administration of SYL-1119.
SYL-1119 是一种用于治疗自身免疫性疾病的鞘氨醇-1-磷酸受体 1 调节剂，具有更好的选择性，而 SYL-1119-P 则是其活性磷酸酯。本文建立并验证了一种灵敏、特异的液相色谱-串联质谱法，用于同时测定大鼠血浆中的 SYL-1119 和 SYL-1119-P。SYL-1110 是 SYL-1119 的类似物，用作内标。采用乙腈沉淀蛋白法处理血浆样品。采用甲醇和含 0.1%甲酸的水作为梯度流动相，在 Zorbax SB-C18 柱（3.5μm，100mm×2.1mm）上进行分离，流速为 0.2ml/min，柱温为 20°C。采用正离子电喷雾串联三重四极杆质谱，以多反应监测模式检测，SYL-1119 的监测离子对为 m/z 364→259，SYL-1119-P 的监测离子对为 m/z 444→259，内标的监测离子对为 m/z 378→273。SYL-1119 的线性范围为 0.2-50ng/ml，SYL-1119-P 的线性范围为 10-1000ng/ml。SYL-1119 的定量下限（LLOQ）为 0.2ng/ml，SYL-1119-P 的定量下限（LLOQ）为 10ng/ml。两种分析物的日内和日间精密度均为 5.4-12.8%，准确度在±10%范围内。两种化合物的回收率均为 91.3-104.5%。结果表明，在所有样品储存、制备和分析过程中，分析物均稳定。该方法成功应用于大鼠口服 SYL-1119 后 SYL-1119 和 SYL-1119-P 的药代动力学研究。
