Department of Pharmaceutical Analysis, Pharmacy School, Shenyang Pharmaceutical University, Shenyang 110016, China.
J Chromatogr B Analyt Technol Biomed Life Sci. 2012 Jun 15;899:81-5. doi: 10.1016/j.jchromb.2012.05.005. Epub 2012 May 8.
Decitabine is chemically unstable at physiological temperature and pH. In addition, the bioanalysis of decitabine is easily interfered by endogenous 2-deoxycytidine. A simple, sensitive and specific LC-MS/MS method was developed for the analysis of decitabine in rat plasma. No exogenous stabilizers were used to prevent the degradation of decitabine in rat plasma. After deproteinized with acetonitrile at room temperature, rat plasma samples were analyzed on a Hypersil APS-2 NH₂ column interfaced with a triple quadrupole tandem mass spectrometer in positive electrospray ionization mode. Decitabine was completely separated from 2-deoxycytidine using gradient elution of water (solvent A) and acetonitrile (solvent B) at a flow rate of 1 mL/min. To quantify decitabine and daidzin (internal standard), respectively, multiple reaction monitoring (MRM) transitions of m/z 251.1→134.7 and m/z 417.3→255.3 was performed. The assay was linear over the concentration range of 5.0-2000 ng/mL. The intra- and inter-day precision was within 12.0% in terms of relative standard deviation (RSD%) and the accuracy within 5.9% in terms of relative error. The LC-MS/MS method was fully validated for its sensitivity, selectivity, stability study, matrix effect and recovery. The data indicate that this LC-MS/MS method is a specific and effective method for the pharmacokinetic study of decitabine in rat plasma. Compared with the previously reported analytical methods, this method showed easy and economic sample preparation, good specificity and high sensitivity with less plasma (50 μL).
地西他滨在生理温度和 pH 值下化学不稳定。此外，地西他滨的生物分析容易受到内源性 2-脱氧胞苷的干扰。本研究建立了一种简单、灵敏、特异的 LC-MS/MS 法用于分析大鼠血浆中的地西他滨。未使用外源性稳定剂来防止大鼠血浆中地西他滨的降解。室温下用乙腈沉淀蛋白后，采用 Hypersil APS-2 NH₂柱，以水（溶剂 A）和乙腈（溶剂 B）为流动相，在正电喷雾离子化模式下，以 1mL/min 的流速进行梯度洗脱，实现了地西他滨与 2-脱氧胞苷的完全分离。分别以 m/z 251.1→134.7 和 m/z 417.3→255.3 进行多重反应监测（MRM）转换，定量分析地西他滨和大豆苷元（内标）。地西他滨的浓度在 5.0-2000ng/mL 范围内呈线性关系。日内和日间精密度以相对标准差（RSD%）表示，均在 12.0%以内，准确度以相对误差表示，均在 5.9%以内。该 LC-MS/MS 方法经过充分验证，具有灵敏度、选择性、稳定性研究、基质效应和回收率。数据表明，该 LC-MS/MS 方法是一种用于大鼠血浆中地西他滨药代动力学研究的特异性和有效方法。与之前报道的分析方法相比，该方法具有简便、经济的样品制备、良好的特异性和较高的灵敏度，所需血浆量较少（50μL）。
