An Improved High-Performance Liquid Chromatographic Assay for the Determination of Pyridinoline in Connective Tissues

Abstract

A high-performance liquid chromatographic (HPLC) method for determination of pyridinoline and its application to connective tissues are described. The chromatographic separation was accomplished by using Inertsil ODS-2 column and a mixture of 0.1M sodium phosphate and acetonitrile (75:25, v/v) containing sodium dodecyl sulfate (SDS) and ethylenediamine tetraacetic acid (EDTA) as eluent. The chromato-gram was developed isocratically and the eluted components were moni-tored with excitation at 295nm and emission at 395 nm. The pyridinoline in crude hydrolysate of connective tissues can be determined in 5 min, with as little as 1 pmol of sample. The present method is rapid, simple and can be used for the routine analysis of connective tissues such as cartilage, bone, Achilles tendon and aorta.