Estimation of the Volume of Blood in a Small Disc Punched From a Dried Blood Spot Card
Estimation of the Volume of Blood in a Small Disc Punched From a Dried Blood Spot Card
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DOI：
10.1002/ejlt.201700362
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发表时间：
2018-03-01
影响因子：
2.7
通讯作者：
Muhlhausler, Beverly S.
中科院分区：
文献类型：
--
作者：
Hewawasam, Erandi;Liu, Ge;Muhlhausler, Beverly S.
专门的干血斑（DBS）收集系统（PUFAcoat）与液相色谱-串联质谱（LC-MS/MS）相结合，能够在最小体积的血液中测量多种分析物。本研究旨在确定从我们的DBS系统中获得的3 mm和6 mm圆盘中的血量。使用四种不同的方法估计从DBS卡获得的3 mm和6 mm圆盘中的血液体积：（i）重量分析;（ii）LC-MS/MS;（iii）血红蛋白比色测定;和（iv）GC。在方法之间比较估计体积的差异，并且确定个体内和个体之间的估计血容量的变化。对于3 mm和6 mm圆盘，DBS圆盘中的平均血液体积分别计算为1.60.4 L和8.71.9 L。该估计值在直接和间接分析方法之间以及在具有独立于该方法的不同起始体积的DBS样本之间相似，但是来自不同个体的相当大小的DBS圆盘中的血液体积存在相当大的差异。目前的方法能够估计从DBS获得的小圆盘中的血量，但需要一种既能准确测量体积又能在DBS上存储血液的方法。这项研究表明，可以使用一系列不同的方法来评估从干血斑（DBS）卡上打孔的小圆盘中所含的血液体积，但从不同个体采集的样本的估计体积存在相当大的差异。如果假定体积固定，这些变化可能导致高估或低估各种代谢物的内源性水平，这可能具有临床意义。虽然有几种方法来校正DBS上包含的血量，但这些方法的实用性和通用性值得怀疑。理想情况下，需要开发一种工具来更好地确定血液体积和/或精确地点出血液体积，以确保在表达每单位血液的DBS分析结果时的准确性。需要知道从DBS获得的3或6 mm圆盘中的血液体积，以使用液相色谱-串联质谱法准确定量每单位血液的内源性代谢物水平。
A specialized dried blood spot (DBS) collection system (PUFAcoat) in combination with liquid chromatography-tandem mass spectrometry (LC-MS/MS) has enabled the measurement of numerous analytes in minimal volumes of blood. The current study aimed to determine the volume of blood in 3 and 6mm discs obtained from our DBS system. The volume of blood in 3 and 6mm discs obtained from DBS cards is estimated using four different methods: (i) gravimetric analysis; (ii) LC-MS/MS; (iii) a hemoglobin colorimetric assay; and (iv) GC. Differences in the estimated volume are compared between methods, and variations in estimated blood volume within and between individuals are determined. The average volume of blood in a DBS disc is calculated to be 1.60.4L and 8.71.9L for 3 and 6mm discs, respectively. This estimate is similar between direct and indirect analytical methods and between DBS samples with different starting volumes independent of the method, but there is considerable variation in the volume of blood in comparably-sized DBS discs from different individuals. Current methods enable the estimation of the blood volume in a small disc obtained from a DBS but a method that can both accurately measure volume and store blood on a DBS is required.Practical Applications: This study demonstrates that it is possible to evaluate the volume of blood contained in a small disc punched from a dried blood spot (DBS) card using a range of different methods, but there is considerable variation in the estimate of volume in samples collected from different individuals. These variations could potentially result in overestimation or underestimation of endogenous levels of various metabolites if presuming a fixed volume, which may be clinically significant. Although there are several ways to correct for blood volume contained on a DBS, the practicality and the universality of these methods are questionable. Ideally, the development of a tool to better determine the volume of blood and/or to precisely spot a volume of blood is required to ensure accuracy when expressing the results of DBS analyses per unit of blood. The volume of blood in a 3 or 6mm disc obtained from a DBS needs to be known to accurately quantify endogenous levels of metabolites per unit of blood using liquid chromatography-tandem mass spectrometry.