Procedure for the determination of biogenic amines by derivatization with dabsyl chloride and reversed phase liquid chromatography

Abstract

SummaryExisting procedures for the determination of biogenic amines by derivatization with dabsyl chloride and separation by RPLC differ very little from those employed in amino acid analysis. Since some difficulties attributable to the higher hydrophobicity of dabsyl-amides were detected on using these procedures, important modifications are proposed, including extraction of the reaction mixture with an organic solvent, drying of the extract and redissolving the residue in acetone before injecting. Dabsylation and chromatographic conditions were also studied. Although recovery from spiked meat extracts is very good, significant matrix effects are detected when extraction is carried out from spiked whole meat samples. These adverse effects can be overcome by means of standard addition.

Dabsyl derivatisation as an alternative for dansylation in the detection of biogenic amines in fermented meat products by reversed phase high performance liquid chromatography

“…20 min at 70°C is recommended (Romero et al, 2000). In contrast to dansylation (which shows severe decomposition of dansylated amines at the temperatures higher than 65°C), Dadáková et al (2009) and Castillo and Castells (2001) have mentioned that at higher temperatures the peak ascribed to the excess of dabsyl reagent decreased, without the loss of the peak areas of the analytes. In this study, we therefore investigated, if dabsylation could be carried out at 80°C in order to reduce the dabsyl chloride peak.…”

“…In contrast to the dansylation (pH between 9.5 and 10.0), according Romero et al (2000) the pH range for the dabsyl reaction has to be set between 8.0 and 8.5, with an optimal value of 8.2. Other authors mention buffering of the reaction solution at pH 8.6 (Krause et al, 1995), and even at 9.2 (Castillo & Castells, 2001). To check, if the pH range for the derivatisation is as critical, as described by Romero, we carried out an investigation in order to determine the pH range for robust derivatisation.…”

“…In that way, interferences from UV-absorbing biological compounds present in the meat extracts are mostly avoided (Aboul-Enein, 2003). For dabsylation of biogenic amines in several foodstuffs, small variations amongst the methods can be found (Castillo & Castells, 2001;Krause et al, 1995;Romero, Bagur, Sánchez-Viñas, & Gázquez, 2000). Therefore this study is carried out to determine the critical parameters affecting the derivatisation yields of the aforementioned BAs.…”