背景:由于其高特异性和敏感性,液相色谱-串联质谱(LC-MS/MS)是治疗药物监测中免疫抑制剂定量的金标准方法。在这种情况下,干血斑(DBS)已成为一种有前途的策略作为样品收集程序。尽管DBS优于静脉穿刺的优势是众所周知的,这种方法的局限性极大地影响了分析结果的接受度.其中,最重要的是血细胞比容(Ht)。克服此问题的最简单方法是分析完整的斑点。在这个战略中,称为纸盘上的干燥基质(DMPD),血液在体积上施加在预打孔的圆盘上。
目的:为了验证LC-MS/MS方法对他克莫司的定量,西罗莫司,依维莫司和环孢菌素A使用DMPD。
方法:该程序根据国际指南使用商业试剂盒进行验证。评估了以下性能参数:选择性,结转,线性度准确度,精度,定量下限,相对回收率,可交换性和稳定性。此外,我们进行了一项方法比较研究,以评估Ht对结果的临床影响.
结果:所有性能参数均在验收标准范围内,因此,经确定,经验证的方法符合预期目的.同样,在医学决策水平上计算出的偏倚值显示,Ht对结果没有临床影响.
结论:与已发布的其他类似方法不同,在这里,一种简单的方法已得到充分验证。这是第一个LC-MS/MS方法,适用于使用DMPD作为采样策略的商业试剂盒。
BACKGROUND: Due to its high specificity and sensitivity, liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) is the gold standard method for immunosuppressant quantification in therapeutic drug monitoring. In this context, dried blood spots (DBS) have become a promising strategy as a sample collection procedure. Although the advantages of DBS over venipuncture are well known, this approach has limitations that strongly influence the acceptance of analytical results. Among them, the most important is hematocrit (Ht). The easiest way of overcoming this problem is by analyzing complete spots. In this strategy, called dried matrix on paper discs (DMPD), blood is volumetrically applied on pre-punched discs.
OBJECTIVE: To validate an LC-MS/MS method for the quantification of tacrolimus, sirolimus, everolimus and cyclosporin A using DMPD.
METHODS: The procedure was validated according to international guidelines using a commercial kit. The following performance parameters were evaluated: selectivity, carryover, linearity, accuracy, precision, lower limit of quantitation, relative recovery, commutability and stability. In addition, a method comparison study was performed to evaluate the clinical influence of Ht on the results.
RESULTS: All performance parameters were within acceptance criteria and, hence, it was determined that the validated method is fit for the intended purpose. Likewise, calculated bias values on medical decision levels showed that there was no clinical influence of Ht on the results.
CONCLUSIONS: Unlike other similar methodologies that have been published, here, a simple method has been fully validated. This is the first LC-MS/MS methodology adapting a commercial kit to use DMPD as a sampling strategy.