Extractability-mediated stability bias and hematocrit impact: High extraction recovery is critical to feasibility of volumetric adsorptive microsampling (VAMS) in regulated bioanalysis
Extractability-mediated stability bias and hematocrit impact: High extraction recovery is critical to feasibility of volumetric adsorptive microsampling (VAMS) in regulated bioanalysis
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DOI：
10.1016/j.jpba.2018.04.001
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发表时间：
2018-07-15
影响因子：
3.4
通讯作者：
Bateman, Kevin P.
中科院分区：
文献类型：
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作者：
Xie, Iris;Xu, Yang;Bateman, Kevin P.
体积吸收微进样（VAMS）是一种新的微进样技术，被评价为支持调控生物分析的潜力。我们对MK-0518（雷替重力韦）进行了初步评估，使用直接提取方法，提取回收率为45-52%，红细胞压积（Ht）相关偏差显著，更重要的是，在7天的储存后，稳定性不可接受（与标称浓度偏差为15%）。我们的研究表明，所观察到的偏差不是由于VAMS的吸收、取样技术、批次间的可变性、基质效应和/或化合物的化学稳定性，而是由于提取回收率低。为了提高测定回收率，改进的液液萃取（LLE）方法表现出更一致的性能，最小的高温影响（高温范围为20%至65%），以及可接受的样品稳定性。同样的策略成功地应用于另一种更亲水的模型化合物MK-0431（西格列汀）。这些结果表明，之前观察到的高温效应和“不稳定性”实际上是由于不一致的提取性，优化提取回收率大于80%是确保VAMS性能的关键。我们建议在调控生物分析中实施VAMS之前，将Ht独立回收率作为可行性评估的一部分，以消除长期可提取性介导的稳定性偏差。(C) 2018 Elsevier B.V.版权所有
Volumetric absorptive microsampling (VAMS), a new microsampling technique, was evaluated for its potential in supporting regulated bioanalysis. Our initial assessment with MK-0518 (raltegravir) using a direct extraction method resulted in 45-52% extraction recovery, significant hematocrit (Ht) related bias, and more importantly, unacceptable stability (>15% bias from nominal concentration) after 7-day storage. Our investigation suggested that the observed biases were not due to VAMS absorption, sampling techniques, lot-to-lot variability, matrix effect, and/or chemical stability of the compound, but rather the low extraction recovery. An effort to improve assay recovery led to a modified liquid-liquid extraction (LLE) method that demonstrated more consistent performance, minimal Ht impact (Ht ranged from 20 to 65%), and acceptable sample stability. The same strategy was successfully applied to another more hydrophilic model compound, MK-0431 (sitagliptin). These results suggest that the previously observed Ht effect and "instability" were in fact due to inconsistent extractability, and optimizing the extraction recovery to greater than 80% was critical to ensure VAMS performance. We recommend adding Ht independent recovery as part of feasibility assessment to de-risk the long-term extractability-mediated stability bias before implementing VAMS in regulated bioanalysis. (C) 2018 Elsevier B.V. All rights reserved.