Comparative interference by haemolysis in automated assays for bilirubin at multiple analyte concentrations

The negative interference caused by haemolysis in manual bilirubin assays contrasts with the positive interference reported for some automated methods utilizing the same basic chemistry. A comparison was therefore made of the haemolysis interference associated with several automated bilirubin methodologies: multilayer film (Kodak Ektachem- total bilirubin (TBil), direct bilirubin (DBil), conjugated bilirubin (Bc), unconjugated bilirubin (Bu),; tableted reagents (Baxter Paramax- TBil, DBil); continuous flow (Technicon SMAC - TBil). Thirty serum pools were analysed (five concentrations of bilirubin, 2-229 mumol/l; six concentrations of haemoglobin, 0.00002-0.052 mmol/1). All methods, except one (Bc), exhibited both positive and negative interference, depending upon the relative haemoglobin and bilirubin concentrations. This interference, at any given haemoglobin concentration, was neither constant nor proportional with increasing bilirubin concentration. These complex patterns of interference suggest that the best characterization of interference is obtained when (1) both percentage-difference and absolute-difference 'interferographs' are plotted; and (2) the interference is determined at multiple analyte concentrations.