Analytical Methods for Quantification of Relative Flying Fish Paste Content in Processed Sea Food (ago-noyaki)

Abstract

Nucleotide sequence and polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis of the 3’-portion of the mitochondrial 16S RNA gene (rDNA) coding sequence was used to authenticate flying fish paste in ago-noyaki. Flying fish paste in ago-noyaki samples was quantified using image analysis of the PCR-RFLP profile. PCR products from standard ago-noyaki samples were digested with AfaI and MunI restriction endonucleases and their electrophoresis patterns were analyzed to produce standard equations for digestion: AfaI digestion: y = 0.0084x + 0.0757, R² = 0.977 and MunI digestion: y = 0.0091x + 0.0153, R² = 0.974. This method was then applied to analyze five and two commercially available ago-noyaki and noyaki products, respectively. The results confirmed proper E-mark labeling of ago-noyaki.