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Title: Determination of Biogenic Amines with HPLC-APCI-MS

item Wu, Ted
item Bechtel, Peter

Submitted to: Meeting Abstract
Publication Type: Abstract Only
Publication Acceptance Date: 12/30/2008
Publication Date: N/A
Citation: N/A

Interpretive Summary:

Technical Abstract: Determination of biogenic amines in fish samples can be used as a quality attribute and are commonly performed using a derivatization step followed by high pressure liquid chromatography (HPLC) and UV detection. Over estimation and misidentification of biogenic amines can occur when interfering compounds elute and absorb at similar wavelengths. A more sensitive and accurate detection method for determining biogenic amines can be achieved with atmospheric pressure using chemical ionization mass spectrometry (ACPI-MS). The objective of this study was to compare the biogenic amine concentrations in fish samples with two detection methods (UV and ACPI-MS) run concurrently after the derivatization step. Samples were blended with 6% trichloroacetic, filter and derivatized with benzoyl chloride. Separation and quantification of seven biogenic amines were conducted on a reverse phase C18 column with a flow rate of 0.8 ml/min on an Agilent 1100 HPLC. The mobile phase was as followed 1:1 methanol:water for 0.5 min, ramped up to 3:1 methanol:water at 7 min and maintained for 3 min then ramped up to 100% methanol and maintained for 2 min. The UV wavelength used for detection was 254 nm. A full mass spectrum scan with APCI was taken of the derivatized biogenic amine standards and the most abundant ion was used in the selective ion mode for detection and quantification. The setting for the mass spectrum was as followed gas temperature = 350 C, vaporizer = 250 C drying gas = 5 l/min nebulizer pressure and capillary current at 4000 V. Results suggested APCI-MS to be a more selective and sensitive detection method. Overlapping peaks with biogenic amines were observed in samples with UV detection and not the case with APCI-MS. Significantly (p<0.05) higher concentrations of putrescine and 2-phenylethylamine were quantified with the UV detection in samples tested. Tryptamine was detected in low concentrations with ACPI-MS, but not detected with UV. The APCI-MS method that was developed in this study is more sensitive and selective than the commonly used UV method.