Berm E J J, Brummel-Mulder E, Paardekooper J, Hak E, Wilffert B, Maring J G
Department of Pharmacy, Section Pharmacotherapy and Pharmaceutical Care, University of Groningen, Antonius Deusinglaan 1, Groningen, 9713AV, The Netherlands,
Anal Bioanal Chem. 2014 Apr;406(9-10):2349-53. doi: 10.1007/s00216-014-7619-9. Epub 2014 Feb 4.
Dried blood spot (DBS) sampling and quantitative analyses of many current therapeutic drug monitoring (TDM)-guided drugs are advantageous because of the minimal invasive sampling strategy. Here, a fast and robust LC-MS/MS method was developed and analytically validated for simultaneous determination of venlafaxine (VEN) and O-desmethylvenlafaxine (ODV) in DBS. Six-millimeter circles were punched out from DBS collected on Whatman DMPK-C paper, and the DBS was extracted with acetonitrile/methanol at 1:3. The total run time was 4.8 min. The assay was linear in the range of 20-1,000 μg/L for both VEN and ODV. Assay accuracy and precision was well within limits of acceptance (LLOQ = 20 μg/L). Normal hematocrit concentrations (0.30-0.50) did not influence the results neither did a normal spot volume (40-80 μL). Punch position at the perimeter instead of the center of the blood spot gave a bias ranging from 2.4 to 10.4%. Correlation between plasma and spiked DBS samples was high. The concentrations found in spiked DBS samples were higher than those in plasma, indicating that a conversion factor for translation of DBS to plasma values is needed. This analytically validated method is suitable for determination of VEN and ODV in DBS and applicable for TDM. The method will be used for TDM of VEN in the Dutch CYSCE multicenter trial (NCT01778907).
由于采用微创采样策略，干血斑（DBS）采样及对许多当前治疗药物监测（TDM）指导用药进行定量分析具有优势。在此，开发了一种快速且稳健的液相色谱-串联质谱（LC-MS/MS）方法，并对其进行分析验证，用于同时测定DBS中的文拉法辛（VEN）和O-去甲基文拉法辛（ODV）。从Whatman DMPK-C纸上收集的DBS中冲出6毫米的圆片，DBS用乙腈/甲醇按1:3进行提取。总运行时间为4.8分钟。该测定法对VEN和ODV在20 - 1000μg/L范围内呈线性。测定的准确度和精密度均在可接受限度内（LLOQ = 20μg/L）。正常血细胞比容浓度（0.30 - 0.50）和正常斑点体积（40 - 80μL）均不影响结果。血斑周边而非中心的打孔位置产生的偏差范围为2.4%至10.4%。血浆与加标DBS样品之间的相关性较高。加标DBS样品中测得的浓度高于血浆中的浓度，表明需要一个将DBS值转换为血浆值的转换因子。这种经过分析验证的方法适用于测定DBS中的VEN和ODV，适用于TDM。该方法将用于荷兰CYSCE多中心试验（NCT01778907）中VEN的TDM。
