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5% HEO SA-HEO-1%: coated with sodium alginate containing 1% HEO The fish fillets were dipped for 30?sec in 500?mL of the each coating solution. Then, the coated fillets stood for 2?min, followed by a second immersion in CaCl2 (Sigma-Aldrich Chemical Co.) [http://www.selleckchem.com/products/azd5363.html AZD5363] for 30?sec to achieve better crosslinking (Lu et?al. 2009). Next, the samples were allowed to drain completely in ambient condition for about 30?min. Finally, they were stored at 4?��?1��C until testing. Chemical and microbiological analyses were performed at 4-day intervals to determine the overall quality of the fish for 16?days. Chemical analysis Proximate composition The moisture content and crude ash were determined in an oven at 103 and 550��C, respectively, until the weight became constant. The total crude protein was determined by Kjeldahl's method (AOAC 2002) and the lipid content was analyzed according to the procedure of Bligh and Dyer (1959). All measurements were repeated three times for studying repeatability. The total volatile basic nitrogen (TVB-N) TVB-N of the bighead carp samples was measured by the micro-diffusion method as described by Goulas and Kontominas (2005). The values were reported in mg N/100?g of fish. Measurements were repeated three times for studying repeatability. Evaluation of lipid oxidation The peroxide value (PV) was expressed in mEq oxygen/lipid and determined in the total lipid extracts according to the method of Pearson (Egan et?al. 1997). Free fatty acid (FFA) was determined by the procedure explained by AOAC (2002) and its content [http://www.selleckchem.com/products/c646.html selleck screening library] was expressed as percentage of oleic acid. The colorimetric [https://en.wikipedia.org/wiki/Chlormezanone Chlormezanone] method described by Kirk and Sawyer (1991) was used to measure the thiobarbituric acid (TBA) value in fish fillet for secondary lipid oxidation products evaluation. All measurements were repeated three times for studying repeatability. Microbiological analysis The pour plate method was used to determine total viable count (TVC) and total psychrotrophic count (TPC). Ten grams of the fish minced sample was aseptically taken and homogenized in 90?mL of sterile 85% NaCl solution with a blender (HBM-400B, HBM Biomed, Tianjin, China) at room temperature. Appropriate dilutions were serially prepared and then 1?mL of each was spread onto plate count agar media (Merck, Darmstadt, Germany). The prepared plates were incubated at 37��C for 2?days for TVC, and at 10��C for 7?days for TPC. All counts were expressed as log colony-forming units (CFU)/g and performed in triplicate. Statistical analysis The differences among all measurements were evaluated by one-way analysis of variance (ANOVA). Duncan's multiple range tests were used to compare the means to identify which groups were significantly different from other groups. Significance was defined at P?