HPLC Determination of Putrescine and Cadaverine in Serum of Cancer Patients Using Acetylacetone as Derivatizing Reagent

Abstract

Liquid chromatographic procedure has been developed for the determination of putrescine (Put) and cadaverine (Cad) after derivatization with acetylacetone in aqueous-methanolic solution at pH 8.2. The derivatives were eluted and separated from p. Bondapak C-18, 10 urn (3.9x300 mm id) with water methanol:acetonitrile (73:22 05 v/v/v) with a flow rate 1.2 ml/min. The UN/ detection was at 310 nm. Linear calibration curves were obtained 25-200 ng Put and Cad / injection (10 pl). The method was used for determination of Put and Cad in the blood serum of cancer patients before, during and after radiotherapy and were in the range of Put 0.622-3.674 S.D 0.017-0.153 ug/mL and Cad 0.229-1.854 ± S.D 0.002-0.046 µs/ML. The limits of detection were observed 2.5 ng Put and Cad / injection (10 pL). the recoveries of Put and Cad from serum were calculated to 94.2-97,3 %. The concentration of Put and Cad observed were lower after radiotherapy.