Validation Study of BioSystems® Y15 Histamine Dehydrogenase Kit for the Detection of Histamine in Fish and Fishery Products: AOAC Performance Tested MethodSM 072001

Background BioSystems has developed a histamine kit for automated procedure, through the use of a Y15 analyzer, for quantification of histamine in fish and fishery products. Objective Validate the method under the specific guidelines of the AOAC Research Institute Performance Tested MethodSM (PTM) program. Method Samples are extracted with boiling water. The enzymatic method is based on histamine dehydrogenase that catalyzes the oxidation of histamine in the presence of an electron mediator that reduces a dye that is measured at 420 nm. The increase of absorbance is proportional to the histamine concentration. Dispensing of reagents and sample, absorbance readings, calibration, and calculation of results are performed automatically in the analyzer BioSystems Y15. Results The linearity ranges from 0 to 200 mg/kg (r2 > 0.99). The LOQ is 10 mg/kg in all the matrixes. Recoveries range from 75 to 107% at concentrations from 5 to 200 mg/kg, with repeatability precision values between 0.8 and 5.5%. Comparisons with the HPLC reference method shows a good correlation. Cross-reactivity of the assay is negligible for all biogenic amines tested except for agmatine (6.3%). Product consistency was verified by validating lot-to-lot variations and variations within the same lot. Shelf life was verified by real-time stability testing during 40 months at 2–8°C. No differences in histamine detection were observed in robustness testing, in which minor changes are introduced to the assay protocol. Conclusions The automated, simple, and rapid BioSystems Y15 Histamine Dehydrogenase Kit has been successfully validated. Highlights The method is qualified for PTM certification No. 072001.