Application of denaturing high-performance liquid chromatography (DHPLC) for the identification of fish: a new way to determine the composition of processed food containing multiple species.
The identification of fish species in transformed food products is difficult because the existing methods are not adapted to heat-processed products containing more than one species. Using a common to all vertebrates region of the cytochrome b gene, we have developed a denaturing high-performance liquid chromatography (DHPLC) fingerprinting method, which allowed us to identify most of the species in commercial crab sticks. Whole fish and fillets were used for the creation of a library of referent DHPLC profiles. Crab sticks generated complex DHPLC profiles in which the number of contained fish species can be estimated by the number of major fluorescence peaks. The identity of some of the species was predicted by comparison of the peaks with the referent profiles, and others were identified after collection of the peak fractions, reamplification, and sequencing. DHPLC appears to be a quick and efficient method to analyze the species composition of complex heat-processed fish products.
Biofortis SAS , 21 rue de La Noue Bras de Fer, 44200 Nantes, France.
Sophie
S
Le Fresne
Milena
M
Popova
Françoise
F
Le Vacon
Thomas
T
Carton
D000818:Animals; D001483:Base Sequence; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D045303:Cytochromes b / Q000235:genetics; D004247:DNA / Q000032:analysis; D005396:Fish Products / Q000032:analysis*; D005399:Fishes / Q000145:classification* / Q000235:genetics; D005511:Food Handling / Q000379:methods; D006358:Hot Temperature; D011489:Protein Denaturation; D017747:Seafood / Q000032:analysis / Q000145:classification*
D004247:DNA; D045303:Cytochromes b