First NameLaura
Last NameNakovich
Emaillnakovic@vt.edu
Supervisor NameDr. Harold M. McNair
UniversityVirginia Polytechnic Institute
CountryUnited States
KeywordsGas Chromatography, Biogenic Amines, Propyl Chloroformate Derivatives, Mass Spectroscopy, Liquid Chromatography, HPLC
Publication DateOct 28, 2015
DegreeMasters
DomainChemistry

Analysis of Biogenic Amines by GC/FID and GC/MS 2003

Abstract

Biogenic amines (particularly histamine in the presence of putrescine and cadaverine) can lead to scombroid food poisoning if levels become too high. Low levels of these biogenic amines occur naturally, but high levels (FDA sets 50 ppm of histamine in fish as the maximum allowable level) can lead to scombroid poisoning.

A number of analytical methods have been employed to analyze biogenic amines, especially High Performance Liquid Chromatography (HPLC). Amines in general are difficult to analyze by Gas Chromatography (GC) due to their lack of volatility and their interaction with the GC column often leading to significant tailing and poor reproducibility. GC methods could be simpler and cheaper than HPLC equipment in some cases. Biogenic amines need to be derivatized before both GC and HPLC analyses. The objective of this research was to develop a relatively fast, reproducible method to derivatize and quantitate biogenic amines in fish at trace levels using GC/FID. The derivatizing reagent used in the experiments was propyl chloroformate, useful for aqueous samples. A method for GC/MS was also developed and used to confirm the identity of six derivatized biogenic amines. To our knowledge no reference spectra for these derivatives has been published.

It was concluded that best results are obtained using a Cold-On-Column (C.O.C.) inlet with a short column (15 meters), thick film stationary phase (ZB-5, 1.00┬Ám df), and with recommendations to cut 40 cm from the inlet end of the column every 25 injections when using C.O.C. Duplicate samples of Atlantic Salmon were analyzed on days 0, 3, and 5. Levels of histamine were below 50 ppm for days 0 and 3, but day 5 showed average levels of 160 pm (cadaverine), 1000 ppm (histamine), and 350 ppm (tyramine). Good precision of six amine stardards at 50 ppm was shown: heptylamine 5.2%, putrescine 5.6%, cadaverine 5.0%, histamine 9.9%, tyramine 5.1%, and spermidine 6.2% RSD.

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