Exogenous interferences with Jaffe creatinine assays: addition of sodium dodecyl sulfate to reagent eliminates bilirubin and total protein interference with Jaffe methods.

BACKGROUND: The study evaluated the impact of interferences on the analytical specificity of three commercial and commonly used creatinine methods (two Jaffe and one enzymatic). METHODS: Manufacturer creatinine methods plus modified methods were tested with the following interferences: spiking serum with bilirubin, albumin, glucose, hemoglobin and lipid, and patient sera with maximum concentrations of bilirubin, 1,090 micromol/l and protein, 117.8 g/l. RESULTS: Hemoglobin, 7.5 g/l and lipaemic with triglyceride concentration of 6.27 mmol/l, did not interfere with all assays. Glucose >33.3 mmol/l increased creatinine recovery for Dimension method. Samples spiked with bilirubin imparted a negative bias for Dimension and Architect methods but imparted a positive bias for Vitros assay. However, using patient sera, negative bias with bilirubin was found for all methods, from which Architect method gave the highest effect (R(2)=0.861), followed by Vitros (R(2)=0.239) and Dimension (R(2)=0.163). Protein provided the positive bias for all creatinine measurements that increased with increasing concentration (R(2) ranging from 0.104 to 0.182, P<0.0001). Addition of sodium dodecyl sulfate (SDS) in alkaline-picrate reagent reduced the effect of bilirubin and protein for kinetic Jaffe method. Although adding potassium ferricyanide was well effective for eliminating negative interference of bilirubin, it was prone to interference from protein. CONCLUSIONS: Endogenous interferences continue to plague creatinine accuracy measurement in both Jaffe and enzymatic methods, and consequentially the estimated glomerular filtration rate. The addition of SDS to the alkaline-pirate reagent was shown to be effective in reducing bilirubin and protein interferences.

Deproteinization of serum: another best approach to eliminate all forms of bilirubin interference on serum creatinine by the kinetic Jaffe reaction.

The negative interference of conjugated, unconjugated, and delta bilirubin on patient serum creatinine determined by the kinetic Jaffe reaction is the unresolved problem. We compared bilirubin interference on thirty patients' serum creatinine obtained from four analyzers, with and without deprotenization before the Jaffe reaction, to the Vitros dry enzymatic method. We found significant negative interference from bilirubin on serum creatinine in all samples directly applied to four wet chemical methods, except the one incorporated with serum blank rate. The negative interferences linearly related to bilirubin concentration. However, bilirubin did not interfere on serum creatinine obtained from all wet chemical methods incorporated with deproteinization process before the reaction. We conclude that deproteinized serum before the reaction is the best approach to eliminate all forms of bilirubin interference on serum creatinine determined by the kinetic Jaffe reaction.