Novilytic Laboratories , West Lafayette, Indiana 47906, United States.
Anal Chem. 2013 Dec 3;85(23):11501-8. doi: 10.1021/ac402735y. Epub 2013 Nov 7.
A rapid plasma extraction technology that collects a 2.5 μL aliquot of plasma within three minutes from a finger-stick derived drop of blood was evaluated. The utility of the plasma extraction cards used was that a paper collection disc bearing plasma was produced that could be air-dried in fifteen minutes and placed in a mailing envelop for transport to an analytical laboratory. This circumvents the need for venipuncture and blood collection in specialized vials by a phlebotomist along with centrifugation and refrigerated storage. Plasma extraction was achieved by applying a blood drop to a membrane stack through which plasma was drawn by capillary action. During the course of plasma migration to a collection disc at the bottom of the membrane stack blood cells were removed by a combination of adsorption and filtration. After the collection disc filled with an aliquot of plasma the upper membranes were stripped from the collection card and the collection disc was air-dried. Intercard differences in the volume of plasma collected varied approximately 1% while volume variations of less than 2% were seen with hematocrit levels ranging from 20% to 71%. Dried samples bearing metabolites and proteins were then extracted from the disc and analyzed. 25-Hydroxy vitamin D was quantified by LC-MS/MS analysis following derivatization with a secosteroid signal enhancing tag that imparted a permanent positive charge to the vitamin and reduced the limit of quantification (LOQ) to 1 pg of collected vitamin on the disc; comparable to values observed with liquid-liquid extraction (LLE) of a venipuncture sample. A similar study using conventional proteomics methods and spectral counting for quantification was conducted with yeast enolase added to serum as an internal standard. The LOQ with extracted serum samples for enolase was 1 μM, linear from 1 to 40 μM, the highest concentration examined. In all respects protein quantification with extracted serum samples was comparable to that observed with serum samples obtained by venipuncture.
一种快速血浆提取技术，可在三分钟内从指尖采血获得的 2.5μL 血浆样本中提取。该血浆提取卡的优点是可以生成一个载有血浆的纸质收集盘，该收集盘可在 15 分钟内风干，并放入邮寄信封中运输到分析实验室。这样就避免了由采血员进行静脉穿刺和在专门的小瓶中采集血液，也避免了离心和冷藏储存的需要。通过将一滴血液施加到膜堆栈上，通过毛细作用将血浆提取出来，从而实现血浆提取。在血浆迁移到膜堆栈底部的收集盘的过程中，通过吸附和过滤的组合去除血细胞。当收集盘充满血浆样本后，将上部膜从收集卡上剥离，然后将收集盘风干。在收集卡之间，收集的血浆体积差异约为 1%，而在红细胞压积水平为 20%至 71%的范围内，体积变化小于 2%。然后从收集盘上提取含有代谢物和蛋白质的干燥样品并进行分析。在与甾族信号增强标签衍生化后，通过 LC-MS/MS 分析定量 25-羟维生素 D，该标签赋予维生素永久正电荷，并将定量下限 (LOQ) 降低到收集盘上的 1pg 维生素；与静脉穿刺样本的液-液萃取 (LLE) 观察到的值相当。使用传统的蛋白质组学方法和光谱计数进行了类似的研究，将酵母烯醇酶作为内标添加到血清中。对于提取的血清样品中的烯醇酶，LOQ 为 1μM，线性范围为 1 至 40μM，是检查的最高浓度。在所有方面，提取的血清样品的蛋白质定量与通过静脉穿刺获得的血清样品相当。
