Abstract
Surimi-based processed products are prone to mislabeling using raw materials that are not in accordance with food safety requirements. There were cases reporting use of toxic fish tissue in commercial seafood products. This study was aimed to identify and determine the raw materials used in various processed surimi products using cytochrome oxidase I (COI) gene marker. The experiment consisted of DNA isolation, DNA amplification using several target primers namely full-length barcodes, mini-barcodes, as well as specific species primers against poisonous puffer fish Lagocephalus lunaris and genetic analysis. The results of bioinformatics analysis revealed that S1 samples were Coryphaena hippurus or mahi-mahi fish, S2 and S3 samples were Nemipterus bathybius or curated fish and CS samples were Evynnis cardinalis or kuro fish. Detection of samples with species specific primers of puffer fish Lagocephalus lunaris with annealing temperatures of 54°C, 57°C, and 60°C showed no DNA bands in the six samples analyzed.
Highlights
Surimi-based processed products are prone to mislabeling using raw materials
The experiment consisted of DNA isolation
The results of bioinformatics analysis revealed that S1 samples were
Summary
Bahan yang digunakan adalah enam produk olahan surimi impor yaitu crabstick (CS), surimi crab claw king (CL), chikuwa (C), dan tiga sampel fish ball dari berbagai merek dengan kode S1, S2, dan S3. Bahan lainnya yaitu kit isolasi DNA (QIAGEN DNeasy Food Mericon, Venlo, Netherlands), kit PCR (KAPA Taq PCR Kits, Kapabiosystems, Wilmington, USA), ddH2O, empat jenis primer (primer spesifik ikan buntal atau puffer fish yang dirancang pada penelitian ini, primer fish universal F1R1/F2R2 (Ward et al 2005), dan primer fish universal mini-barcode (Shokralla et al 2015), marker DNA (VC 100 bp plus DNA ladder, VIVANTIS, Selangor Darul Ehsan, Malaysia), cybergreen, agarosa (SeaKem®LE Agarose, Lonza, Rockland, ME USA), buffer TBE (AccuGENETIM 10X TBE Buffer, Lonza, Rockland, ME USA). Alat-alat yang digunakan yaitu microtubes TUBE-150-C (Extragene, Taichung, Taiwan) dan PCR-02-C (Axygen, California,USA), pipet tips RC-10/20-L dan RC-250/20-C (Mettler Toledo, Bekasi, Indonesia), micropipette 2-10 μL, dan 20-200 μL (ThermoFisher Scientific, Vantaa, Finland), freezer, waterbath sonicator (BANDELIN electronic, Berlin, Germany), vortex (V1-Plus, Biosan, Warren, USA), spindown (Corning, New York, USA), sentrifugasi (Centurion Scientific 2041, Libertyville, USA), microwave (Sharp, Osaka, Japan), timbangan digital (PGW 254, ADAM®, England), PCR (Applied Biosystem GeneAmp PCR System 9700, TermoFisher Scientific, Vantaa, Finland), UV-Transilluminator (Uvitec, Cambridge, England), elektroforesis (HU6, SCIE-PLAS, Cambridge, England)
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