Comparison of the Vanadate Oxidase Method with the Diazo Method for Serum Bilirubin Determination in Dog, Monkey, and Rat

Abstract

Abstract. The most widely used method for bilirubin concentration determination is the diazo method, which measures the color of azobilirubin. The vanadate oxidase method is based on oxidation of bilirubin to biliverdin by vanadate. The objective of this study was to compare total and direct bilirubin concentration ([Bt] and [Bd], respectively) determined by the diazo and vanadate oxidase methods in pooled serum samples from dogs, monkeys, and rats spiked with panels of different concentrations of bilirubin standards. Pooled serum samples from 40 dogs, 40 monkeys, and 60 rats were spiked with either ditaurine conjugates of bilirubin or a standard reference material. The results obtained from both assays were compared using Deming regression analysis. The intra-and interassay precision, expressed as a percentage of the coefficient of variation (%CV), was determined for [Bt] and [Bd], and the mean percentage of recovery was calculated. The vanadate oxidase method displayed an excellent correlation (r 5 0.99-1.00) with the diazo method. Using Deming regression, there were minimal negative or positive constant and proportional biases for [Bt] and [Bd]. The precision studies revealed that the vanadate oxidase method has comparable between-run and within-run CVs to those of the diazo method. The recovery study demonstrated that the diazo method more closely approximates the expected values of [Bt]. In conclusion, the vanadate oxidase method is a simple and rapid method that can be employed as an alternative to the diazo method when interfering substances are present in the serum samples of dog, monkey, and rat.

Haematobiochemical Profile in Chronic Acetamiprid Exposure in Sprague Dawley Rats

“…It might be due to reduced feed and water intake which was found in present experiment (Ghofrani et al, 2010;Ameri et al, 2011).…”

Efficient detection of a biomarker for infant jaundice by a europium(iii)-organic framework luminescence sensor

“…To date, a variety of available methods including diazo, 9 enzymatic, 10 vanadate oxidation, 11 electrochemical 12 and fluorometric techniques 13 have been consequently applied to recognize bilirubin in human serum and urine samples. Most of these approaches are time-consuming, involve complicated pre-treatment and depend on expensive instruments.…”

A Gold Nanoclusters Film Supported on Polydopamine for Fluorescent Sensing of Free Bilirubin

“…As concerns fBR detection in serum samples, various methods can be found in the reported literature. The classical methods are usually based on diazo reactions or vanadate oxidation [7], but the reaction is indirect (needing an accelerator substance) and pH-dependent. Methods using bilirubin oxidase, peroxidase, or an artificial enzyme have been developed for electrochemical detection of fBR [8,9].…”