Comparison of high pressure liquid chromatography methods in fish and fish products by using o-phthaldialdehyde and benzoyl chloride derivatization for the determination of histamine levels

In this study, by the parameters of validation a comparison of two high pressure liquid chromatography (HPLC) methods one of which utilizes coupled the FLD and the o-phthaldialdehyde derivatization method of Antoine et al. (1999) versus DAD coupled, benzoyl chloride derivatization method with a modification of Ozdestan and Uren (2009) were made for the selection of sensitive and safe assay for the determination of histamine. Alive horse mackerels caught from the sea of Marmara in Kucukcekmece Istanbul were used for the limit of detection studies. For the other validation parameters, recovery, accuracy, precision and comparison of methods a certified reference material; FAPAS T2775QC were used. Results of the analyses indicated that the FLD coupled; o-phthaldialdehyde derivatization method the limit of quantification (LOQ) and recovery were 1.38 ppm and 100% respectively; and for DAD coupled; benzoyl chloride derivatization modified method these values were 10.66 ppm and 88% respectively. There was a significant difference between for both methods in t-test (p<0.5). According to the calculations of Horrat value; the reproducibility was normal for o-phthaldialdehyde method but found to be higher than the accepted limits in benzoyl chloride method. In conclusion, it is advised to choose FLD coupled o-phthaldialdehyde derivatization method for the safe and sensitive determination of histamine in fish and fish products.