Preparation Of Nitrosohemoglobin And Its Application In Meat Products

This project defined the development of the substitute of nitrite in the meat product from color fixative with excellent stability,coloring stability,natural safety,highly antioxidant and antimicrobial activityThe dye can not only lead the meat product to show right color but also can support the stability of product color,lead the meat produce to have slow fading speed,suitable color developing consequence and excellent balance,distinctly enhance the color of meat products,reduce residual nitrite quantity and prolong shelf life of meat productsThe research focused on preparing new type curing pigmentwhich is called NO-Hb.NO-Hb was prepared by the use of by-products(beef blood)and its application prospect substitutes of nitrite is able to increase color stability and inhibit pathogenic bacteria.Optimization conditions used for NO-Hb extractions were pH,VC,NaNO2,and speed 6.0,6.0 g/L,3.5 g/L and 8000 r/min,respectivelyMinced beef with NO-Hb was significantly higher than control(P<0.05),IC50 is a concentration of minced beef with NO-Hb which can inhibit 50%of DPPH anions,the IC50 of minced beef with NO-Hb was 10.53 mg/mL,while the IC50 of vitamin C was 9.84 ?g/mL The ABTS-SA of minced beef with NO-Hb and vitamin C was significantly higher than control(P<0.05).IC50 of minced beef with NO-Hb for the ABTS radical scavenging activity was 10.02 mg/mL while the ABTS radical scavenging activity IC50 of vitamin C was 9.88 ?g/mL at refrigerator storage at 4? for 1-wk.Clearing the minced beef with NO-Hb possesses more significant DPPH inhibition than controlAll bacteria,including pathogenic and non-pathogenic bacteria such as E.coli(E.coli 0157:H7)ATCCB A A-1883,Enterotoxigenic[E.coli(ETEC)ATCC354017,Enterohemorrhagic[E.coli(EHEC)ATCC51657GFP7,Staphylococcus aureus(S.aureus)ATCC31888,Staphylococcus argenteus(S.argenteus)JABA32044V6S1,Staphylococcus simiae(S.simiae)CCM7229(DQ127902),Salmonella Typhi(S.Typhi)ATCC19430,Salmonella Javiana(S.Javiana)ATCC10721,Salmonella Enteritidis[S.Enteritidis ATCC31194,Streptococcus thermophiles(S.thermophilus)ATCC19258,Lactobacillus bulgaricus(L.bulgaricus)ATCC11842,Lactobacillus casei(L.casei)ATCC393,Montessori enterococcus(M.enterococcus)NFR17393,and Pseudomonas lundensis(P.lundensis)ATCC49968,Pseudomonas mudicolens(P.mudicolens)ATCC4685,Pseudomonas aeruginosa(P.aeruginosa)ATCC35554,were evaluated.The effect of NO-Hb on individual pathogenic microbial counts(log10 CFU/g)(1:1)injected in a fresh beef showed that the mean Log10 CFU/g value of total microbial count(TBC),E.coli O157:H7,E.coli(EHEC),and L.casei were at 10-1 Log CFU/g:0.58 ± 0.01,0.12,0.02,and 0.25,respectively in a fresh beef before injection with bacteria.Staphylococcus,Pseudomonas,Salmonella,L.bulgaricus,M.enterococcus,and S.thermophilus were not detected in a fresh beef before injection with bacteria.The control samples bacteria with the ratio(1:1)injected in fresh beef(without NO-Hb addition),all bacterial counts were uncountable(Un),except that L.casei and S.thermophiles appeared at 10-1 Log CFU/g:2.14 ± 0.42,1.58 ± 0.02,respectively.After NO-Hb addition,all bacterial counts were significantly decreased(P<0.05)compared with the control group.The TBCs were 2.02±0.06 at 10-4 Log CFU/g before adding NO-Hb.However,TBCs were significantly(P<0.05)decreased to reach 0.68 at 10-2 log CFU/g after adding NO-Hb.The combination of E.coli,Staphylococcus,and LAB were at 10-2 Log CFU/g:2.27 ±0.01,and 2.18 ±0.03,respectively,before adding NO-Hb.However,E.coli,Pseudomonas,and salmonella growth were rarely observed when combined with LAB and adding NO-Hb.Pseudomonas and LAB counts were at 10-4 Log CFU/g:1.64 ± 0.01 and 0.40,respectively,after 7 d of cold-storage.TBC and LAB were significantly decreased after adding NO-Hb(P<0.05).The combination of LAB itself was more resistant to NO-Hb,compared with other pathogenic.From the finding in this study,it was seen that the intensity of the LAB was lower than that before adding NO-Hb.The red color of the minced beef with NO-Hb was significantly(P<0.05)higher than that of either control or the samples inoculated with LAB.NO-Hb with VC,Ca,and ginger are significant color enhancers and pathogenic inhibitors.Moreover,the functional NO-Hb offered additional functionalities such as the nutritional and safe quality.The supplementation of Ca g/k:1.5,2,2.5,3 with 1 g VC could improve cooking,redness,and storage stability of mince beef.Furthermore,the use of NO-Hb combined with LAB might be useful for substituting nitrate to inhibit the pathogenic organisms.A total of 55 different components were identified by analyzing the volatile components by HS-SPME-GC/MS.The volatile compounds of minced beef with NO-Hb and Arabic gum(GuA)5 g/kg of each had significantly increased in frozen/refrozen at-20? for 3-wk.Generally,55 different compounds were found totally in all different samples.In which,by using similarity clustering test up to 51.37%were divided into five groups.In which they found extracted compounds belong to different chemical classes:aldehydes,alcohols,acids,ketones,esters,hydrocarbon,and phenolic compound.Furthermore,in this study,some of these compounds appeared,increased,decreased or absent after using of NO-Hb+GuA with the minced beef.Low field(LF)-NMR method was established to be practicable to identify changes in the relaxation times of water and fat caused by different types of bacteria and the storage periods.The relaxation time T2b,T21,and T2 were significantly(P<0.05)different among treatments freeze-thaw at-20? for 7 d.The increasing storage was observed to change both the(T2's)and the(A2's)of the water in mined minced beef with NO-Hb+GuAEssential and non-essential amino acid compositions were analyzed using high-performance liquid chromatography(HPLC).Ash content was determined by furnace at 550?for 24 h.Fatty acids were analyzed using gas-liquid chromatography(Shimadzu,GC-14A)The vitamin's content were determined according to the measures of AOAC(2000)by a reversed-phase high-performance liquid chromatography(RP-HPLC)In general,the minced beef with NO-Hb+GuA had high amino acids(AAs)contents in this study.Moreover,NO-Hb+GuA effects onto minced beef microstructure by scanning electron micrographs,fluorescent micrographs and reflection confocal scanning laser microscopy have its benefits to confirm and visualize the changes in minced beef new structures in different conditions.Scanning results showed that the microscopic morphology of the minced beef with NO-Hb(5 g/kg)of each presented a relatively homogeneous structure,whereas in mince beef with NO-Hb and Arabic gum(5 g/kg)of each was more homogeneous with the surface covered by small aggregates and possibly originated by changing in protein interactions and fat distributionMinced beef with NO-Hb 5 g/kg,vitamin C 1 g/kg,calcium g/kg:1.5,2.0,2.5,3.0,and Arabic gum 5.0 g/kg studied was used to prepare minced beef product.The color stability,highly antioxidant,antimicrobial,functional properties,physicochemical properties,nutrition value,volatile components and microstructure of minced beef products were evaluated.In the color evaluation,the NO-Hb with ginger,Arabic gum group was significantly(P<0.05)higher than the control.In the overall score,the NO-Hb with ginger,Arabic,vitamin C and calcium scored the highest than the controlNO-Hb that was integrated with LAB,Vitamin C,calcium,ginger and GuA as a novel ingredient,produced an acceptable pink color and inhibited pathogenic bacteria,which were consistent to the effect of nitrite and could be used as a substitute of nitrite in red meat industry field.