Pharmacogenomics Laboratory, Centre Hospitalier Universitaire de Québec (CHUQ) Research Center, Canada.
J Chromatogr B Analyt Technol Biomed Life Sci. 2012 Feb 15;885-886:131-7. doi: 10.1016/j.jchromb.2011.12.029. Epub 2012 Jan 9.
A liquid chromatography-tandem mass spectrometry method was developed for the quantification of circulating levels of multiple immunosuppressant drugs including cyclosporine (CsA), tacrolimus, methotrexate (Mtx), prednisone, prednisolone, methylprednisone, total and free mycophenolic acid (MPA), as well as MPA phenolic (MPAG) and acyl (AcMPAG) glucuronide metabolites. Linearity, precision and accuracy were validated within the typical therapeutic range of concentrations for each compound. The assay was linear over 0.125-25ng/mL for tacrolimus, 1-500ng/mL for prednisone/methylprednisone, 2-400ng/mL for Mtx, 2-1000ng/mL for prednisolone and from 7.5 to 1500ng/mL for CsA with the lowest limit of quantification (LLOQ) being 0.125, 1.00, 2.00, 2.00 and 7.5ng/mL, respectively. The calibration curve concentrations for MPA and MPAG ranged from 50 to 50,000ng/mL (LLOQ: 50ng/mL) and 10 to 10,000ng/mL (LLOQ: 10ng/mL) for AcMPAG. Mean recoveries in blood and plasma were 84%±5.7%. The method could measure individual drugs with high sensitivity, accuracy (bias≤14%), and reproducibility (CV≤12.8%). Its clinical application was validated by measuring levels of these drugs in samples obtained from hematopoietic stem cell transplant recipients treated with combined immunosuppressive drug therapy. Our results indicate that this approach is suitable for simultaneous determination of in vivo levels of immunosuppressive drugs commonly used in combined therapies.
建立了一种液相色谱-串联质谱法，用于定量检测环孢素（CsA）、他克莫司、甲氨蝶呤（Mtx）、泼尼松、泼尼松龙、甲基泼尼松龙、总游离麦考酚酸（MPA）以及 MPA 酚（MPAG）和酰基（AcMPAG）葡萄糖醛酸代谢物等多种免疫抑制剂药物的循环水平。该方法在每种化合物的典型治疗浓度范围内验证了线性、精密度和准确性。对于他克莫司，该测定法在 0.125-25ng/mL 范围内呈线性，对于泼尼松/甲基泼尼松龙，在 1-500ng/mL 范围内，对于 Mtx，在 2-400ng/mL 范围内，对于泼尼松，在 2-1000ng/mL 范围内，对于 CsA，从 7.5 至 1500ng/mL 范围内呈线性，最低定量限（LLOQ）分别为 0.125、1.00、2.00、2.00 和 7.5ng/mL。MPA 和 MPAG 的校准曲线浓度范围分别为 50 至 50,000ng/mL（LLOQ：50ng/mL）和 10 至 10,000ng/mL（LLOQ：10ng/mL）对于 AcMPAG。血液和血浆中的平均回收率为 84%±5.7%。该方法可以以高灵敏度、准确性（偏差≤14%）和重现性（CV≤12.8%）测量单个药物。通过测量接受联合免疫抑制药物治疗的造血干细胞移植受者样本中这些药物的水平，验证了该方法的临床应用。我们的结果表明，该方法适用于联合治疗中常用的免疫抑制剂药物的体内水平的同时测定。
