Centre for Biospectroscopy, Monash University, Clayton, 3800, Victoria, Australia.
Department of Analytical Chemistry, University of Valencia, Burjassot, Spain.
Anal Methods. 2021 Dec 9;13(47):5756-5763. doi: 10.1039/d1ay01242h.
Blood screening is a fundamental part of disease diagnosis and monitoring health. Attenuated total reflectance Fourier transform infrared (ATR-FTIR) spectroscopy offers an innovative solution to streamlining the process, especially for multianalyte detection in aqueous samples. However, samples always undergo a storage phase before they are processed for testing and blood transfusion. In this study, we investigated the effect of standard storage procedures on the macromolecular composition of whole blood, and plasma collected in blood tubes for diagnostic purposes and initial screening of blood products. Periphery blood samples were collected from 10 volunteers and then stored for 14 days at 4 °C. Samples were stored as isolated plasma and whole blood to provide three different datasets, namely: (1) plasma stored independently, (2) plasma stored with other blood components and (3) whole blood. ATR-FTIR spectra of aqueous blood were acquired every 24 h from the time of collection on a portable ATR-FTIR spectrophotometer to monitor the evolution of the macromolecular composition in each blood component. Principal component analysis (PCA), partial least squares regression (PLS-R) and multi-curve resolution alternate least squares (MCR-ALS) models were built to study changes in the spectra with the storage time and identify the key bands. Isolated plasma stored without red blood cells (RBCs) showed no changes over the 14 day period indicating limited degradation. By contrast, plasma stored with the other blood components showed visual and spectroscopic signs of degradation including increasing lipid bands and the amide I and II bands from haemoglobin (Hb). Ideally, for the application of IR spectroscopy in blood diagnostics and for initial screening of blood products, whole blood and isolated red blood cells can be stored for a maximum of 4 days at 4 °C in lithium-heparin anticoagulant tubes prior to spectral analysis before any signs of degradation. Isolated plasma, on the other hand, can be stored for much longer periods and shows no evidence of degradation in the spectra after 14 days.
血液筛查是疾病诊断和监测健康的基本组成部分。衰减全反射傅里叶变换红外（ATR-FTIR）光谱为简化该过程提供了一种创新的解决方案，特别是在水相样品中的多分析物检测方面。然而，在对样本进行测试和输血之前，它们通常要经历一个储存阶段。在这项研究中，我们研究了标准储存程序对用于诊断目的和血液制品初步筛选的血液管中采集的全血和血浆的大分子组成的影响。从 10 名志愿者中采集外周血样本，然后在 4°C 下储存 14 天。样本分别以单独的血浆和全血的形式储存，以提供三个不同的数据集，分别为：（1）单独储存的血浆，（2）与其他血液成分一起储存的血浆，（3）全血。从采集时开始，使用便携式 ATR-FTIR 分光光度计每 24 小时采集一次水相血液的 ATR-FTIR 光谱，以监测每个血液成分中大分子组成的演变。使用主成分分析（PCA）、偏最小二乘回归（PLS-R）和多曲线分辨交替最小二乘（MCR-ALS）模型来研究光谱随储存时间的变化，并确定关键波段。未储存红细胞（RBCs）的单独血浆在 14 天期间没有变化，表明降解有限。相比之下，与其他血液成分一起储存的血浆出现了视觉和光谱降解迹象，包括脂质带增加以及血红蛋白（Hb）的酰胺 I 和 II 带。理想情况下，对于在血液诊断中的应用以及对血液制品的初步筛选，在进行光谱分析之前，全血和分离的红细胞可以在 4°C 下用锂肝素抗凝管储存最多 4 天，在此期间应无降解迹象。另一方面，单独的血浆可以储存更长的时间，并且在 14 天后的光谱中没有显示出降解的迹象。
