Determination of Biogenic Amines in Fresh Fish and Processed Fish Products Using IC-MS/MS

Abstract

A new method was proposed for the determination of underivatized biogenic amines based on ion-exchange chromatography coupled with mass spectrometry detection. The method was applied to the analysis of 10 biogenic amines in fresh and processed fish products. The amines were extracted from muscle tissue with water without any additional derivative step or sample clean-up. Separation of biogenic amines was done by the IonPac (4 × 50 mm) column, applying a gradient eluent by mixing formic acid (2 mol L−1) and Milli-Q water (formic acid concentration from 400 mM to 2 M). The results demonstrated a linear response in the range of 0.01 to 10 mg L−1. The detection limits for the fish products ranged from 20 ng/g up to around 400 ng/g for histamine and putrescine, respectively. Spermidine and spermine showed significantly higher detection limits. This current method can be used for the determination of biogenic amines in both fresh and processed fish products for regulatory purposes and monitoring food-safety issues relating to these amines, particularly histamine. It is also a useful method for evaluation of other commercial analytical test kits and commonly used methods that are possibly affected by the food matrix due to processing or other drawbacks arising from the derivatization process.

Analysis of Biogenic Amines Using Immunoassays, HPLC, and a Newly Developed IC-MS/MS Technique in Fish Products—A Comparative Study

“…Histamine is a decomposition product of histidine and formed by decarboxylation activity during the growth of certain bacteria in some fish species. The amount of the amine that forms depends on bacterial species, the temperature and exposure time, and may exceed even over 1000 mg/kg [ 2 , 27 ]. Fish containing high histamine levels has been associated with many examples of HFPs which is a major health problem for consumers.…”

“…However, FDA guidelines suggest using 17 mg/kg histamine for HACCP plans to avoid the histamine health risk [ 9 ]. For an effective HACCP plan in the fish processing plant, rapid histamine testing is necessary for monitoring, since histamine and other BAs can continue to form even at a chilled temperature; therefore, the time to take action is very crucial [ 2 , 27 ]. Commercial test kits based on immunoassays are often used for such purposes either for quantitative or semi-quantitative analysis.…”

“…Quantitative methods such as chromatographic techniques are generally used for both regulatory and monitoring purposes while qualitative methods such as enzyme immunoassays are only used for a rapid screening test for histamine only to be applied in the seafood companies’ hazard analysis critical control point (HACCP) systems for monitoring and prevention purposes. These methods have been compared by taking the factors of sensitivity, linearity, rapidity, repeatability, efficiency, and operational requirements into consideration, and advantages and disadvantages have been discussed by various authors over the years [ 2 , 15 , 16 , 17 , 18 , 19 , 20 , 21 , 22 , 23 , 24 , 25 , 26 , 27 ]. For regulatory purposes, reference methods are preferred by the government authorities.…”

“…So far, a few methods have been developed for the analysis of BAs in different foods such as meat, cheese and wine [ 33 , 34 , 35 , 36 , 37 ]. Recently, we have developed an ion chromatography-mass spectrometric detection (IC-MS/MS) method for analyzing BAs in cheese, fish and fisheries products which applies the direct application of the sample extract for reading BAs in these products without derivatization [ 26 , 27 ].…”

“…The earlier studies demonstrated that the IC-MS/MS method showed a good recovery for various BAs in cheese and fish products. The method showed high reducibility and sensitivity, and low detection limits (with the exception of spermidine and spermine) in determining BAs in fish and cheese [ 26 , 27 ]. So far, few quantitative test kits have been validated by the AOAC Research Institute Performance Tested Method SM program, namely, HistaSure™ ELISA (Fast Track) also called Histamine FOOD EIA (by Labor Diagnostika Nord, Nordhorn, Germany, (#021402) and enzyme immunoassay (EIA) (by Kikkoman Biochemifa Company, Tokyo, Japan) Certificate (#042802), and the BioSystems ® Y15 Histamine Dehydrogenase kit (#072001) (by BioSystems S.A., Barcelona, Spain) [ 29 , 31 , 38 ].…”