Determination and Identification of Bilirubin in Jaundiced Pig Liver by High Performance Liquid Chromatography

KAI-CHUN CHANG, MEI-YUN CHEN, KE-JIAN CHENG AND SHIH-PING CHEN *

Animal Technology Institute Taiwan, Miaoli, Taiwan, R.O.C.

(Received: June 17, 2011; Accepted: October 25, 2011)

ABSTRACT

A high performance liquid chromatography (HPLC) method was developed for the determination of bilirubin concentration in pig liver. The sample was extracted by sequential extraction with dimethyl sulfoxide and acetonitrile, followed by HPLC analysis. The accuracy of the analytical method was estimated by spiking pig liver samples with three different concentrations of bilirubin (2.5, 5 and 10 μg/mL) and a recovery of 75.7 - 84.9% was obtained, with corresponding intra-day and inter-day precisions of 1.6 - 4.1% and 2.4 - 3.9%, respectively. The limit of detection of bilirubin was 0.3 μg/mL and the analytical time for one sample was 9 min. In this study, the bilirubin concentration of normal and jaundiced pig livers was determined. It was concluded that the procedure could provide scientific data for detecting increased bilirubin concentration to identify jaundiced livers. The analytical method was found to be rapid, sensitive, accurate, repeatable and reproducible, and could be applied for the measurement of bilirubin concentration in pig liver.

Key words: bilirubin, jaundice, high performance liquid chromatography