Development of a new HPLC method for rapid histamine quantification in fish and fishery products without sample clean-up

HPLC methods are among the most studied and validated techniques for the quantification of histamine in fish and fishery products; nonetheless, most of these methods require laborious sample extraction and purification. For this purpose, in the present study, an HPLC-FLD method without any sample clean-up procedure was developed, validated and applied for histamine quantification in various fish samples. Histamine was extracted from fish and fishery products samples with perchloric acid; then a simple precolumn derivatization with o-phthalaldehyde was performed in the presence of the reducing agent 2-mercaptoethanol. Once the derivatization was carried out, the separation was performed with a reverse phase HPLC system. In terms of validation, good linearity and specificity over a concentration range from 12.5 to 200 mg/kg were achieved. Also, convenable precision with relative standard deviations smaller than 5.92%, accuracy ranged from 0.40 to 1.84%, and satisfactory recoveries ranged from 100.44 to 101.84%, were obtained. The detection and the quantification limits were calculated at 2.8 and 4.3 mg/kg, respectively. In addition, the effectiveness of the method was evaluated by the analysis of reference materials, proficiency tests and various type of samples. Moreover, the developed method was found to be simple, rapid, and reliable.

Automated summary

An HPLC-FLD method without sample clean-up procedure was developed for quantifying histamine in fish and fishery products. The method demonstrated good linearity, specificity, precision, accuracy, and satisfactory recoveries within a certain concentration range. It was found to be simple, rapid, and reliable, effectively evaluated by various tests and sample analyses.