• Title/Summary/Keyword: fish detection

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Development and Validation of Multiplex Polymerase Chain Reaction to Determine Squid Species Based on 16s rRNA Gene (오징어류 종 판별을 위한 다중 유전자 검사법 개발 및 검증)

  • Kim, Hyunsu;Seo, Yong Bae;Choi, Seong-Seok;Kim, Jin-Hee;Shin, Jiyoung;Yang, Ji-Young;Kim, Gun-Do
    • Journal of Food Hygiene and Safety
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    • v.30 no.1
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    • pp.43-50
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    • 2015
  • In this study, single PCR and multiplex PCR tests were examined for identification of four types of squid species (giant squid, cuttlefish, octopus, beka squid) purchased from fish market as well as aquatic processed products in Busan. To design the specific primers against each species, the nucleotide sequences of the mitochondrial 16s rRNA gene of Architeuthis dux, Todarodes pacificus, Enteroctopus dofleini, Enteroctopus megalocyathus, Uroteuthis chinensis, Uroteuthis duvauceli, Uroteuthis edulis groups were analyzed for the identification of each species registered in the GeneBank (www.ncbi.nlm.nih.gov) and have been used for comparative analysis. In order to obtain the size variation of amplified fragments on multiplex PCR, we designed KOJ-F, OJ-F, OCT-F, HAN-F, ALLR primers for each species. The optimal PCR conditions and primers were selected for four types of squid species to determine target base sequences in its PCR products. In the case of single PCR, giant squid was only amplified by KOJ-F/ALLR primer; cuttlefish was only amplified by OJ-F/ALLR primer; octopus was only amplified by OCT-F/ALLR primer; and beka squid was only amplified by HAN-F/ALLR primer. For multiplex PCR, the mixture of four kinds of genomic DNA (giant squid, cuttlefish, octopus, beka squid) been prepared as a template and used together with the mixture of KOJ-F/OJ-F/OCT-F/HAN-F/ALLR primers in the reaction. By the multiplex PCR, it is confirmed that four samples are correspond to multiple simultaneous amplicon. Finally, we validated the established methods of multiplex PCR in the aquatic processed products. Although the mitochondrial 16s rRNA primers used in this study was useful as a marker for detection of each species among them, the study indicated that the established multiplex PCR method can be more useful tool for monitoring the processed products.

A Study on the Analytical Method of Artificial Sweeteners in Foods (식품 중 인공감미료의 분석법에 관한 연구)

  • Kim, Hee-Yun;Yoon, Hae-Jung;Hong, Ki-Hyung;Lee, Chang-Hee;Park, Sung-Kwan;Choi, Jang-Duck;Choi, Woo-Jeong;Park, Sun-Young;Kim, Ji-Hye;Lee, Chul-Won
    • Korean Journal of Food Science and Technology
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    • v.36 no.1
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    • pp.14-18
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    • 2004
  • Analysis methods of artificial sweeteners, aspartame, acesulfame potassium, sodium saccharin, and sucralose isolated from foods were developed using high performance liquid chromatography, HPLC conditions for aspartame, acesulfame potassium, and sodium saccharin were: column, Symmetry $C_{18}(3.9mm\;i.d{\times}150mm,\;5{\mu}m)$; mobile phase, 0.05M sodium phosphate monobasic : acetonitrile (9 : 1, pH 3.5, containing 0.01M tetrapropylammonium hydroxide); detector, UV detector at 210 nm. HPLC condition for sucralose were : column, Symmetry $C_{18}(3.9mm\;i.d{\times}150mm,\;5{\mu}m)$; mobile phase, water:methanol (7 : 3); detector, refractive index detection (sensitivity = 16). Recoveries of artificial sweeteners in foods including soft drinks, fruit and vegetable beverages, alcoholic beverages, fermented milk beverages, soybean milk, ice cream, snacks, chewing gums, jam, honey, kimchi salted food, special dietary products, processed fish products, candies, food additive mixtures, chocolate and cocoa were 76.1-101.3%, 82.3-103.2%, 83.1-103.7%, and 80,6-99.5% for aspartame, acesulfame potassium, sodium saccharin, and sucralose, respectively.