A novel calorimetric reagent for the detection of biogenic amines.

摘要

The development of methods for the detection and quantitation of chemical compounds in matrix samples continues to be an important aspect of analytical chemistry. The determination of biogenic amines in foods and food products is an important application due to the role of these amines in spoilage which constitutes a potential health hazard. A novel solid-phase based derivatization method is reported herein for the detection of biogenic amines in foods and specifically in fish. The method makes use of an off-line red colored visualization reagent which upon reaction with amines, derivatizes them resulting in a red color dye tagged amines. HPLC followed by absorbance detection was used for the validation of the method. Using solid-phase synthetic techniques, polymer-supported 1-hydroxybenzotriazole (P-HOBT) was obtained by the coupling reaction of 3-aminopropyl silica gel with 1-hydroxybenzotriazole-6-carboxylic acid (HOBT-COOH). The resulted P-HOBT was activated by the reaction of P-HOBT with 3,4-dihydroxy-9,10-dioxo-anthracenesulfonyl chloride (alizarin red S chloride) resulting in the off-line red colored visualization reagent. The reagent was reacted with aliphatic and aromatic amines which lead to the derivatization of these amines and consequently to the formation of sulfonamides. The loading capacity of P-HOBT activated with alizarin red S chloride was typically 0.20-0.26 mmol/g. P-HOBT was also activated with acetic anhydride, and the resulted off-white activated P-HOBT was an amine derivatizing reagent. The reagent was treated with aliphatic or aromatic amines which lead to the derivatization of these amines and the formation of amides. The loading capacity of P-HOBT activated with acetic anhydride was typically 0.40-0.45 mmol/g. In the application of P-HOBT activated with alizarin red S chloride to the detection of biogenic amines and volatiles amines in foods, mackerel, a fish from Scombridae family, was used as a model fish for the study. This is due to the fact that fish from the Scombridae family typically contain high concentrations of biogenic amines during spoilage. In this study, biogenic amines such as histamine, putrescine, tyramine, volatile amines such as diethylamine, and aliphatic amine such as pyrrolidine were detected. Putrescine, tyramine and diethylamine were quantified in the conditions of the assays. The results showed that on day 0 mackerel obtained from a local supermarket contained 2 ppm of tyramine and 1.8 ppm of diethylamine. During the 14 days storage, the concentrations of these amines at -15 degrees did not change significantly, whereas at 37 degrees a significant increase was observed reaching concentrations maximum of 166 ppm (day 14) for tyramine, 27.4 ppm (day 7) for diethylamine, and 112 ppm (day 14) for putrescine. These results showed that the off-line red colored visualization reagent was capable of the detection of biogenic amines before spoilage (50 ppm) and during spoilage (≥50 ppm) making the reagent a valid candidate for the detection of spoilage in fish.