Cell lysing solutions are essential reagents used by hematology analyzers to categorize and enumerate cell types such as erythrocytes (RBC), thrombocytes (platelets), and leukocytes (WBC) in whole blood. Lysing reagents are not easily pre-assessed for strength before formulation and use in hematology assays. Lytic strength depends on the chemical properties and the concentration of the lytic agent. The relationship between the lytic strength and the hemolysis rate has been mostly an experimental practice in laboratories and hasn’t been well characterized. Here, a first order kinetics model was hypothesized for the hemolysis by chemical detergent. The kinetics study of the hemolysis was conducted with a control blood sample and a lytic agent: sodium dodecyl sulfate (SDS). The course of hemolysis was monitored on a UV-Vis spectrophotometer, which measured absorption. However, the absorption measure comes from RBC particles scattering incident light, which decreases as the number of particles diminishes. The lysate was monitored at the wavelength of 700 nm to avoid potential interference from hemoglobin absorption. Concentration of RBC was fixed and lytic agent concentrations were varied in the experiment. The data profile yielded the time course of the absorption value. After data analysis, a linear relationship of 1/T<sub>half</sub> and [Lys] was confirmed to validate the hypothesis of the first order kinetics. The slope of 1/T<sub>half</sub> vs [Lys] was found to be 1.44k, which was further explored as a kinetics parameter to enumerate the lytic strength of the lysing reagents in general.

Determination of a quantitative indicator of the lytic strength of cell lysing reagents

2019 Fall National Meeting

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