Assessing the matrix effects of hemolyzed samples in bioanalysis

Validation of LC-MS/MS assays includes an assessment of matrix effects. Hemolysis effect, a special type of matrix effect, can also have an impact on analyte quantitation. In situations where the hemolysis effect is marginal, this can be resolved simply by dilution of hemolyzed samples with plasma prior to analysis. However, in some cases, the impact can be so dramatic that analytes are completely immeasurable. In such situations, modification to the bioanalytical method will be required, including, but not limited to, adjusting the chromatographic conditions to separate interferences present in hemolyzed samples; additional sample clean-up techniques such as protein precipitation in combination with SPE or a change in extraction technique such as from SPE to a liquid-liquid extraction method. Here, we report examples from four bioanalytical methods, where the presence of hemolyzed blood in plasma was found to have an impact on analyte quantitation and a description of the solutions adopted to resolve this are provided.