Development of an automated on-line pre-column derivatization procedure for sensitive determination of histamine in food with high-performance liquid chromatography–fluorescence detection

• Published in: Journal of Chromatography A 2008-10, Vol.1209 (1-2), p.70-75
• Main Authors: Peng, Jin-feng, Fang, Ke-teng, Xie, Dong-hua, Ding, Bin, Yin, Ju-Yi, Cui, Xiao-mei, Zhang, Ying, Liu, Jing-fu
• Format: Article
• Language: English
• Subjects: aldehydes; Automated on-line pre-column derivatization; automation; Biological and medical sciences; cheeses; chemical analysis; chemical reactions; Chromatography, High Pressure Liquid - methods; detection; fish; fish products; fluorescence; Food; food analysis; Food Analysis - methods; Food industries; Fundamental and applied biological sciences. Psychology; General aspects; high performance liquid chromatography; Histamine; Histamine - analysis; Histamine - chemistry; HPLC–FLD; Methods of analysis, processing and quality control, regulation, standards; o-phthaldialdehyde; Reproducibility of Results; Spectrometry, Fluorescence - methods
• ISSN: 0021-9673
• DOI: 10.1016/j.chroma.2008.09.028

An improved sensitive method was developed and validated for the determination of histamine in food samples by using automated on-line pre-column derivatization coupled with high performance liquid chromatography and fluorescence detection (HPLC–FLD). o-Phthaldialdehyde (OPA) was adopted as derivatization reagent, and a “sandwich” (OPA+histamine+OPA) aspiration mode for the automated on-line pre-column derivatization was found to efficiently enhance the method sensitivity and precision. Histamine in food samples was efficiently extracted with a methanol–phosphate buffer solution (50:50, v/v) at 60°C for 30min, and purified with Waters Oasis MCX solid-phase extraction (SPE) cartridge. The limit of quantification for this method is 0.2mg/kg, which is very sensitive for histamine determination. With the “sandwich” injection program, 3.7% of relative standard deviation (RSD) was achieved by five replicative determinations of a sample blank spiked with 0.25mg/kg histamine standard. Histamine in food samples such as fumitory skipjack and mackerel was analyzed with relative recoveries over 95% at spiking level of 150mg/kg, as well as canned tuna fish and cheese with relative recoveries up to 98% at spiking levels of 0.50 and 5.0mg/kg, respectively. The proposed method was validated with a sample from the Food Analysis Performance Assessment Scheme (FAPAS) as a standard certified material; and the results (140±6mg/kg) agreed well with the assigned value (139mg/kg).