The optimal method to measure polyamines in serum by using HPLC fluorescence detector

Abstract

Measurement of polyamines (putrescine, spermidine and spermine) is important in the monitoring of many metabolic events, as well as in cancer prognosis. However, the lack of method standardization and validation for the measurement of poly-amines is a major obstacle. In this study, sample preparation, gradient program, chromatography column, sample concentration studies were attempted and an analysis method was developed for the detection of polyamines in serum by using HPLC fluorescence detector in order to reduce the difficulties experienced with the existing methodologies. This method was validated analytically in accordance with EMA and FDA’s guidelines. In this study, Waters Nova-Pak C18 A chromatography column with 3.9 mm, 150 mm and 4 µm specifications was used for the first time. Acetonitrile was used instead of methanol. Thanks to the newly developed gradient, peak purity and sufficient peak separation were ensured and accurate, sensitive, reliable and reproducible method was developed and validated.