Journal of Chromatography B, 745 (2000) 431–437 www.elsevier.com / locate / chromb Short communication Rapid and simultaneous high-performance liquid chromatography assay of polyamines and monoacetylpolyamines in biological specimens * G. Taibi , M.R. Schiavo, M.C. Gueli, P. Calanni Rindina, R. Muratore, C.M.A. Nicotra University of Palermo, Institute of Biological Chemistry, Via Del Vespro 129, 90127 Palermo, Italy Received 20 December 1999; received in revised form 10 April 2000; accepted 22 May 2000 Abstract A rapid, resolutive and reproducible reversed-phase high-performance liquid chromatography (RP-HPLC) method was developed for polyamines and acetylpolyamines by adopting pre-column derivatization with benzoyl chloride. In a single run lasting less than 15 min ten polyamines were separated as well as traces of benzoic acid, methylbenzoate and benzoic anhydride. These contaminants, produced during the derivatization reaction, were almost all eliminated by washing steps envisaged in the same procedure. This simple and sensitive method can be applied to routine determination of polyamines in biological samples. A fine application of this procedure to the determination of endogenous content of polyamines in chick embryo retina was reported.  2000 Elsevier Science B.V. All rights reserved. Keywords: Polyamines; Monoacetylpolyamines 1. Introduction that the intracellular concentration of these acetylated forms varies greatly, according to the proliferative Polyamines are present in significant amounts in status of the cell and also in response to a wide range both prokaryotic and eukariotic cells. Their role in of stimuli [8–10]. High concentrations of the regulation of main functions in cell growth and acetylpolyamines have been found in chick embryo cell differentiation has drawn the attention of many retina proliferation and development [11,12] and in researchers. Their involvement in DNA replication, malignant breast tissues, whereas they are not detect- 1 gene expression, protein synthesis and cell surface able in normal tissues [13]. N -Acetylspermidine has receptor functions has been widely accepted [1–5]. also been identified as an excretion product of Over-production of polyamines can be cytotoxic to cultured cells [14], in which acetylation seems to be cells [6] or, in the presence of negative growth a prerequisite for excretion [15,16]. signal, can induce apoptosis [7]. Acetylpolyamines The availability of a sensitive assay for the are also present in cells and it has been ascertained simultaneous detection of polyamines and their conjugated forms is important to detect the fluctua- tions of the levels of these metabolites related to the *Corresponding author. Tel.: 139-91-655-2472; fax: 139-91- regulation of cell proliferation and differentiation. 655-2457. E-mail address: gtaibi@unipa.it (G. Taibi). Several studies have been performed on the HPLC 0378-4347 / 00 / $ – see front matter  2000 Elsevier Science B.V. All rights reserved. PII: S0378-4347(00)00314-5