Thalidomide measurement in plasma and dried plasma spot by SPE combined with UHPLC-MS/MS for therapeutic drug monitoring: Supplementary files

Aims: To validate an SPE-ultra-HPLC-MS/MS method for thalidomide (THD) measurement in dried plasma spot (DPS). Methods: Extraction included acetonitrile/water clean-up and online SPE. The LOD, LLOQ, linearity, precision, accuracy, recovery, matrix effect, process efficiency, carryover, stability, drug interference and dilution integrity were assessed. Results: The method was linear from 50 to 2000 ng/ml with a LOD of 20 ng/ml and LLOQ of 50 ng/ml. The coefficient of variation for precision was 0.4–7.9% for intra-assay and 1.3–8.9% for interassay and accuracy was 81.4–97.1%. Adequate matrix effect (100.6–107.0%), recovery (88.7–105.0%) and process efficiency (91.3–109.3%) were registered. DPS was stable for 14 days at room temperature and 45◦C and for 4 months at -80◦C. The method was applied to quantify THD in both wet plasma and DPS from patients with cutaneous lupus receiving THD treatment. The difference between THD wet plasma and DPS concentration was suitable to quantify THD in DPS.

研究目的：验证一种用于干血浆斑（dried plasma spot，DPS）中沙利度胺（thalidomide，THD）检测的固相萃取-超高效液相色谱串联质谱（SPE-ultra-HPLC-MS/MS）方法。实验方法：采用乙腈-水溶液净化结合在线固相萃取的提取方案，对该方法的检出限（limit of detection，LOD）、定量下限（lower limit of quantitation，LLOQ）、线性范围、精密度、准确度、回收率、基质效应、过程效率、残留效应（carryover）、稳定性、药物干扰及稀释完整性进行了系统性评估。研究结果：该方法在50~2000 ng/ml浓度范围内呈现良好线性关系，检出限为20 ng/ml，定量下限为50 ng/ml。批内精密度的变异系数为0.4%~7.9%，批间精密度变异系数为1.3%~8.9%，准确度为81.4%~97.1%。基质效应为100.6%~107.0%，回收率为88.7%~105.0%，过程效率为91.3%~109.3%，各项指标均符合分析方法验证要求。干血浆斑样本在室温及45℃条件下可稳定保存14天，在-80℃超低温条件下可稳定保存4个月。本方法已应用于接受沙利度胺治疗的皮肤狼疮患者的湿血浆及干血浆斑样本中沙利度胺的定量检测，湿血浆与干血浆斑中的沙利度胺浓度差值满足干血浆斑样本定量检测的适用性要求。