Three direct spectrophotometric methods for determination of total bilirubin in neonatal and adult serum, adapted to the Technicon RA-1000 analyzer

We adapted three bichromatic spectrophotometric methods for determining total bilirubin in serum, for use with the Technicon RA-1000 analyzer. The borate buffer (BOR) of Hertz et al. (Scand J Clin Invest 1974;33:215-30), the caffeine buffer (CAF) of Vink et al. (Clin Chem 1988;34:67-70), and the combined borate-caffeine buffer (B-C) of Franzini and Cattozzo (Clin Chem 1987;33:597-9) were compared. All methods required only 10 microL of serum, were precise (between-batch CVs less than 4.2%, analyte range 64-310 mumol/L), linear to 1000 mumol/L, and insensitive to interference from hemoglobin to 5 g/L. Lipemia, carotene, and methemalbumin interfered positively with each method but insignificantly unless in supranormal concentrations. Only the BOR method was sensitive to protein matrix effect. Neonates' results obtained with all three methods compared well with a Jendrassik-Gróf-based technique. However, samples from adults with cholestasis were overestimated, particularly by the CAF method, but the BOR and B-C methods would be suitable for "stat" bilirubin analysis in these samples.