VERIFICATION AND ASSESSMENT OF THE MEASUREMENT UNCERTAINTY OF THE HPLC METHOD FOR DETERMINING THE AMOUNT OF HISTAMINE IN FISH AND FISHERY PRODUCTS

Abstract

The aim of the study is to present the experimental results obtained during the verification of the HPLC method for determining the histamine content in fish and fishery products for three types of fish and fishery products and to estimate measurement uncertainty of the method. The method for determining the amount of histamine in fish and fishery products is verified according to BAS EN ISO 19343:2017. The following parameters of the verification method were evaluated: influence of the matrix/linearity, limit of quantification, accuracy and precision. Measurement uncertainty of the method was estimated for three types of fish (tuna, mackerel and sardines). Linearity was confirmed in the concentrations range from 0 mg kg-1 to 500 mg kg-1 with correlation coefficients from 0.9925 to 0.9999. The limit of quantification for all three types of fish is 10 mg kg-1. The accuracy was evaluated by analyzing the control materials through recovery and the average value is 96.61%. The precision was evaluated in terms of repeatability and reproducibility at three concentration levels for three types of fish and was expressed as the repeatability limit r and the reproducibility limit R. The reproducibility limits R for tuna are 15.96 to 24.89%, for mackerel 12.80 to 23.04%, and for sardines from 10.25 to 32.17%. The measurement uncertainty was estimated at three concentration levels 25 mg kg-1, 100 mg kg-1 and 220 mg kg-1 and was 17.78 mg kg-1, 11.40 mg kg-1 and 13.88 mg kg-1 for tuna, 16.46 mg kg-1, 12.56 mg kg-1 and 9.14 mg kg-1 for mackerel and 22.98 mg kg-1, 12.70 mg kg-1 and 7.32 mg kg-1 for sardines. The obtained results of the verification parameters are in accordance with the prescribed and recommended acceptance criteria, and confirm that the test method is sufficiently reliable and the laboratory can use it in routine work.