Veterynarna biotehnologija – Veterinary biotechnology, 2019, 35, 62-70 [in Ukrainian]. https://doi.org/10.31073/vet_biotech35-08
State Scientific and Research Institute of Laboratory Diagnostics and Veterinary and Sanitary Expertise
National University of Life and Environmental Sciences of Ukraine
VALIDATION OF THE METHOD FOR DETERMINING HISTAMINE IN FISH AND FISH PRODUCTS BY THE METHOD OF LIQUID CHROMATOGRAPHY USING AN UV DETECTOR AND DERIVATIZATION
Introduction. Histamines are biogenic amines that are formed in fish and fish products during the decarboxylation of histidine with the involvement of enzymes that produce microflora with impaired storage conditions. Histamine is dangerous for humans in high concentrations, causing a number of diseases that even lead to death. As a result, there is a need to introduce a reference method of high performance liquid chromatography for the determination of histamine in fish and fish products with derivatization and the use of internal standards. Validation data and validation data are presented in detail.
The goal of the work was to implement a confirmatory method for determining the mass concentration of histamine in fish and fish products and assess the suitability of the method for compliance with specific requirements.
Materials and methods. Materials for research were river and sea fish. Studies were carried out on a liquid chromatograph Varian LC-1. The principle of the method is to extract the sample, carry out the derivatization of dansyl chloride, determine the mass concentration of histamine in the ratio of the area of external standards to the area of the internal standard.
Results of the research and their discussion. The assessment of the appropriateness of the method was carried out in accordance with the error rates specified in Commission Regulation 2002/657/EC of 12 August 2002 on the implementation of Council Directive 96/23/EC on the application of analytical methods and the interpretation of results. The method has high metrological characteristics: repeatability from 2.8% to 5.6%, reproducibility from 3.2% to 9.1%, trueness from 91% to 99%.
Conclusions and prospects for further research. The technique introduced by us allows us to detect histamine in fish and fish products using a liquid chromatograph using an ultraviolet detector within 10-250 mg/kg. The methodology has been successfully tested, complies with the requirements of Commission Regulation 2002/657/EC on the implementation of the Council Directive 96/23 on the application of methods and interpretation of results and accredited by the National Accreditation Agency of Ukraine.
Keywords: histamine, fish, high performance liquid chromatography (HPLC), allergy, 1.7-diaminoheptane (DAH), internal standard, external standards, dansyl chloride, derivatization.