Department of Chemistry and Biochemistry, Concordia University, Montreal, Quebec, Canada H4B 1R6.
Anal Biochem. 2013 May 15;436(2):66-8. doi: 10.1016/j.ab.2013.01.024. Epub 2013 Jan 31.
As erythrocyte-derived extracellular adenosine triphosphate (ATP) gains recognition as a key vasodilator, its accurate determination is critical. Erythrocytes' high hemoglobin content can act as an inner filter when measuring ATP concentrations by chemiluminescence. We evaluated two approaches to correct for this matrix effect: addition of cell-free hemoglobin to the ATP standards and standard addition of ATP to erythrocyte suspensions. In addition, we reduced sample hematocrit to minimize the absorbance. We conclude that extracellular ATP should be determined in erythrocyte suspensions at 0.06 to 0.004% hematocrit. This gives robust signals without matrix effects and requires only microliters of blood.
随着红细胞衍生的细胞外三磷酸腺苷（ATP）作为一种关键的血管扩张剂得到认可，其准确测定至关重要。当通过化学发光法测量 ATP 浓度时，红细胞的高血红蛋白含量可作为内滤光器。我们评估了两种方法来校正这种基质效应：向 ATP 标准品中添加无细胞血红蛋白和向红细胞悬浮液中标准添加 ATP。此外，我们降低了样品的血细胞比容，以最大程度地减少吸光度。我们得出结论，应在红细胞悬浮液中以 0.06%至 0.004%的血细胞比容测定细胞外 ATP。这可以在没有基质效应的情况下产生稳健的信号，并且仅需要微升血液。
