Determination Of Biogenic Amines In Fish Based Products Using Liquid Chromatographic And Capillary Electrophoresis
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Date
2018-04
Authors
Bakar, Padilah
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Publisher
Universiti Sains Malaysia
Abstract
Determination of biogenic amines (BA) in fish based products were carried out using two standard methods, reversed phase - high performance liquid chromato- graphy/UV-Vis (RP-HPLC/UV-Vis) and capillary electrophoresis with capacitively coupled contactless conductivity detection (CE-C4D). The objective of the study was to determine the level of BA in local fish based products for safety assurance using standardised method yet sensitive, simple and at low cost. Using RP-HPLC method, putrescine (PUT), cadaverine (CAD), histamine (HIS) and tyramine (TYR) were analysed in 15 local belacan products. About 53% of belacan products contain HIS above 50 mg kg-1 safe limit (means, 57.63 mg kg-1) but below 200 mg kg-1 ‘rejected level’ for quality assessment in fermented fisheries products for international market. Salt content (NaCl) are above 15% in belacan products and moisture percentage are less than 40% in 80% of belacan samples, under the regulation for belacan products in Food Act 1983/Food Regulation 1985. The RP-HPLC/UV-Vis optimised method was sensitive and accurate with qualitative detection limit (LOD) of 0.01 μg mL-1 and quantitative detection limit (LOQ) ranging from 0.04 - 0.05 μg mL-1 for CAD, PUT, HIS and TYR. However, HPLC method using UV-Vis detector required expensive derivatisation reagent and laborious procedures. Hence, a new method using CE-C4D was developed as alternative for BA analysis in food. Eight BA namely HIS, PUT, CAD, TYR, spermine (SPM), spermidine (SPD), tryptamine (TRP) and 2-phenylethylamine (PHE) were determined using a CE-C4D method in one analysis with direct detection within 19 minutes. Method optimisation for eight BA separation were obtained using 0.4 M d-malic acid as background electrolyte, pH 2.11, temperature 24 oC with applied voltage of 20 kV; C4D input signal at frequency 600 kHz and 50% amplitude.
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Keywords
Determination of biogenic amines in fish based products , using liquid chromatographic and capillary electrophoresis