Rat brain polyamines: an analytical method validation

Jornal Brasileiro de Patologia e Medicina Laboratorial

Valdomiro De Freitas Sampaio²  Gilberto J. Padovan¹  Julio Sérgio Marchini¹  Luiz Marcellino De Oliveira¹  Sebastião Sousa Almeida¹ 

[1] ,Universidade de São Paulo Ciências e Letras de Ribeirão Preto Faculdade de Filosofia

关键词: Validation;    Analytical technique;    HPLC;    Polyamines;    Validação;    Técnica analítica;    HPLC;    Poliaminas;   

DOI  :  10.1590/S1676-24442010000600005

来源: SciELO

【 摘 要 】

The validation of the analytical technique for the determination of polyamines in cerebral tissue using HPLC based on o-phthalaldehyde post-column derivatization is described. The polyamines were separated in a LiChrospher100 RP18 column. Elution gradient was formed with two mobile phases: A (sodium acetate 0.1 M + sodium octanesulphonate 0.01 M, pH = 4.5) and B (sodium acetate 0.2 M + sodium octanesulphonate 0.01 M)/acetonitrile (10:3), pH = 4.5) in a 1.2 ml/min flow rate. The derivative eluent was monitored by fluorescence (excitation, 345 nm; emission, 455 nm). Besides excellent linearity (putrescine, r = 0.9816; spermidine, r = 0.9920; spermine, r = 0.9901), the technique demonstrated intra and inter-day precision (< 20%) as well as recovery (spermidine = 92.56%; spermine = 84.47%). Quantification limits were 0.22 pM for putrescine, 76.44 pM for spermidine and 51.44 pM for spermine. The method demonstrated to be robust, simple and highly reproducible for polyamine determination in tissues.

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