Why should hemolyzed serum be avoided for total protein analysis?

Hemolyzed serum should be avoided for total protein analysis primarily because hemoglobin released from red blood cells can interfere with the absorbance measurement in assays. Hemolysis occurs when red blood cells break apart, releasing hemoglobin into the serum. This hemoglobin can absorb light at the wavelengths used for measuring total protein, leading to falsely elevated results. The presence of hemoglobin creates additional color in the serum that contributes to a nonspecific increase in absorbance readings, thus skewing the analytical results for total protein.

While alterations in pH, protein denaturation, or increased viscosity can occur with hemolyzed samples, the most significant impact on total protein measurement specifically comes from the interference of hemoglobin with the assay's absorbance. This makes it crucial to ensure that serum samples are not hemolyzed before conducting total protein assays to obtain accurate and reliable results.