T/GDBK 007-2023 Rapid determination of acrylamide in baked foods using enzyme-linked immunosorbent assay (fluorescent probe method)

Scope

Scope: This document specifies the principles, reagents and materials, instrumentation, sample preparation, experimental procedures, data processing, precision, and test reports for the rapid determination of acrolein content in baked goods by fluorescence. This document is applicable to the determination of acrolein in baked goods. The method has a detection limit of 1.12 μg/kg and a quantification limit of 3.92 μg/kg. Main technical content: 4. Principle: The sulfhydryl group in N-acetyl-4-mercapto-1,8-naphthalimide has a PET effect on the fluorescent nucleus of naphthalimide, and its own fluorescence is very weak. However, in the presence of acrolein, the sulfhydryl group of N-acetyl-4-mercapto-1,8-naphthalimide reacts with acrolein through the Michael addition reaction to form an adduct, which inhibits the PET effect. The adduct emits strong fluorescence under excitation at a wavelength of 380 nm and has a peak at a wavelength of 510 nm. Under fixed conditions, the fluorescence intensity is proportional to the concentration of acrolein in the sample. The determination is carried out using an enzyme analyzer with a fluorescence detector and quantified by external standard method. The principle of fluorescence probe method for determining acrolein is shown in Figure 1. Figure 1: Principle of fluorescence probe method for determining acrolein 5. Reagents and materials 5.1 Reagents: Unless otherwise specified, all reagents are of analytical grade, and the water is Grade I water specified in GB/T 6682. 4-bromo-1,8-naphthalic anhydride (analytical grade). Glycine (analytical grade). Triethylamine (analytical grade). Ethanol (analytical grade). Hydrochloric acid (analytical grade). P-toluenesulfonic acid (analytical grade). Methanol (analytical grade). Sodium sulfide nonahydrate (analytical grade). Dimethylsulfoxide (DMSO, analytical grade). Citric acid (analytical grade). Dinatriumphosphate (analytical grade). Acrolein (analytical grade). 5.2 Preparation of fluorescen

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