评估并确定用于干血斑液相色谱-串联质谱分析的样品制备程序。
Evaluating and defining sample preparation procedures for DBS LC-MS/MS assays.
作者信息
Liu Guowen, Patrone Laura, Snapp Heidi M, Batog Alicja, Valentine Jack, Cosma Greg, Tymiak Adrienne, Ji Qin C, Arnold Mark E
机构信息
Bioanalytical and Discovery Bioanalytical Sciences, Bristol-Myers Squibb Co., Rte 206 and Province Line Road, Princeton, NJ 08543, USA.
出版信息
Bioanalysis. 2010 Aug;2(8):1405-14. doi: 10.4155/bio.10.106.
BACKGROUND
A defined approach to develop and validate an LC-MS/MS assay using dried blood spot (DBS) samples is of great interest to many scientists who are adopting this technology. We have evaluated three distinct sample preparation procedures of DBS samples for LC-MS/MS assay development.
RESULTS
A new term 'elution efficiency' is introduced to evaluate the effectiveness of eluting compounds from the DBS cards into the liquid phase. Three different types of DBS cards were studied as part of the sample preparation procedures. A DBS LC-MS/MS method was developed, qualified and then applied to a toxicokinetics study.
CONCLUSION
Organic extraction and protein precipitation resulted in significant ion suppression and/or enhancement for FTA(®) Classic or FTA(®) Elute cards. Liquid-liquid extraction produced the least ion suppression/enhancement. Both protein precipitation and liquid-liquid extraction effectively eluted the probe compound from the DBS cards under the conditions tested. However, organic extraction by pure solvents resulted in low elution efficiency.
背景
对于许多采用该技术的科学家而言，开发和验证一种使用干血斑（DBS）样本的液相色谱-串联质谱（LC-MS/MS）测定方法的明确方法具有重大意义。我们评估了用于LC-MS/MS测定方法开发的三种不同的DBS样本制备程序。
结果
引入了一个新术语“洗脱效率”来评估从DBS卡中将化合物洗脱到液相中的有效性。作为样本制备程序的一部分，研究了三种不同类型的DBS卡。开发并验证了一种DBS LC-MS/MS方法，然后将其应用于毒代动力学研究。
结论
对于FTA(®) Classic卡或FTA(®) Elute卡，有机萃取和蛋白质沉淀会导致显著的离子抑制和/或增强。液-液萃取产生的离子抑制/增强作用最小。在测试条件下，蛋白质沉淀和液-液萃取均能有效地从DBS卡中洗脱探针化合物。然而，使用纯溶剂进行有机萃取会导致洗脱效率较低。
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