The Influence Of Curcumin In Residue And Eliminate Of Afb1 And Its Metabolite AFM1 In Liver, Kidney And Muscles Of Broiler

Aflatoxins(Aflatoxins,referred to as AFT),is a group of fungal secondary metabolites produced by the Aspergillus species,among them aflatoxin B1(AFB1)is the most toxic one and it is widely distributed in moldy food and its products.Its in vivo metabolism will produce aflatoxin M1(AFM1),also has a strong toxicity,and both of them will remain in the animal's liver,kidney,muscle and other edible parts,become a direct threat to human health,so to provide a high sensitivity method to detect aflatoxin residue is essential;Curcumin is the main active substance of traditional Chinese medicine turmeric,has a certain protective effect to human body,and studies have shown that curcumin on aflatoxin poisoning has a protective effect,but still not clear its mechanism of action,and its effect on AFB1 and its metabolite AFM1 residues is still unknown.There are many kinds of methods for the detection of aflatoxin.But mot many people use ultra-high performance liquid chromatography.Ultra-high performance liquid chromatography(UHPLC)is better than high performance liquid chromatography(HPLC),Compared with HPLC,the sensitivity and separation of UHPLC have been greatly improved,The detection time is greatly shortened,the use of solvent is reduced and the detection cost is minimized.Therefore,the establishment of an accurate,sensitive and efficient ultra performance liquid chromatography for the detection of aflatoxin residues in edible tissues of broilers is necessary.And it has a great significance to ensure the safety of livestock and human health.In this study,an ultra-high performance liquid chromatography-fluorescence detector(UHPLC-FLD)was developed to detect AFB1 and AFM1 residues in the liver,kidney and muscle tissues of broilers.The chromatographic and pretreatment conditions were optimized.C18(1.7?m × 2.1mm × 50mm)of Acquity UPLC BEH column was used.The mobile phase was acetonitrile: water(2: 8)and flow rate was 0.2 m L/min.The injection volume was 10 ?L.The column temperature was 25 ° C;The excitation wavelength was 365 nm and the emission wavelength was 435 nm.Select dichloromethane for the pre-treatment extraction solution,and the AFM1 immunoaffinity column was used for purification and derivative approach select trifluoroacetic acid pre-column derivatization.The methodology test results showed that adding three different concentrations of aflatoxin B1 and M1 in blank tissue.The recovery of AFB1 were 82.32-85.56%,85.34-88.45% and 84.88-89.73% respectively in the liver,kidney and muscles,AFM1 average recovery were 92.17-95.03%,94.12-97.21% and 95.32-98.51% respectively.The limits of detection for AFB1 and AFM1 were 0.008 ng/mL and 0.003 ng/m L respectively.The limit of quantification was 0.02 ng/mL and 0.01 ng/m L respectively.The detection method of AFB1 and AFM1 in broilers meat with excellent recovery,sensitivity,minimum toxin lose,and can be used for the quality control testing of commercial broiler-meat processing companies,food manufacturers and control laboratories to check the quality of meat to ensure food safety and avoid health problems.In the present study,the AFB1 and AFM1 residues in AFB1 subchronic poisoned broilers were analyzed.The animals were divided into two groups.The model group was treated with aflatoxin B1(5 mg/kg)for 28 days.The curcumin intervention group The broilers were fed with aflatoxin B1(5 mg/kg)+ curcumin(300 mg/kg)for 28 days.After 28 days,the broilers were fed with standard feeds.Broilers were killed at 28,30,32,35,38 and 42 days and collect the tissues.The residual values of AFB1 and AFM1 in the liver,kidney and muscle were detected by the established detection method.The results showed that the AFB1 residues of different tissues from high to low was muscle,kidney and liver,With the intervention of curcumin,the AFB1 residues in the three tissues increased.The AFB1 metabolites AFM1 residues of different tissues from high to low was kidney,liver and muscle,and with the intervention of curcumin,AFM1 residues were reduced.The residual elimination time of AFB1 and AFM1 was calculated by WT1.4 software according to the residual value of different sampling time.Since there is no AFB1 and AFM1 standard MRLs in tissues,the residual elimination time of AFB1 was calculated according to the LOQ and LOD of AFB1 in the establishment method.The results showed that use LOQ of AFB1 as MRL,the clearance time calculated for the liver,kidney and muscle tissues of the model group were 11,15 and 18 days respectively,Curcumin intervention group,the clearance time of liver,kidney and muscle tissue were 17,19 and 17 days respectively.Using LOD of AFB1 as MRL,the clearance time calculated for the liver,kidney and muscle tissues of the model group were 18,21 and 25 days respectively,Curcumin intervention group,the clearance time of liver,kidney and muscle tissue were 21,25 and 22 days respectively.Since the residual value of AFM1 in the tissue is low,only the residual of 28 d was higher than LOQ,indicating that if use LOQ as MRL,the residual elimination time of AFM1 is less than two days;Using LOD of AFM1 as MRL,the clearance time calculated for the liver,kidney and muscle tissues of the model group were 18,21 and 25 days respectively,Curcumin intervention group,the clearance time of liver,kidney and muscle tissue were 21,25 and 22 days respectively.In summary,In this study,the ultra-high performance liquid chromatography-fluorescence detector(UHPLC-FLD)method(developed and validated)was used for the simultaneous detection of AFB1 and AFM1 residues in the liver,kidney and muscle tissues of broiler chickens.The method was highly sensitive and reproducible,which provides a new method for AFB1 and AFM1 residual detection.From the detection of AFB1 and AFM1 residues in liver,kidney and muscle of broiler chickens in AFB1 subchronic poisoning model group and curcumin intervention group,It was found that curcumin could inhibit the metabolism of AFB1.The residual elimination time of AFB1 and AFM1 from AFB1 subchronic poisoning broiler tissue was analyzed,The results of the study to protect food safety and the development of AFB1 residual effective control measures have important guiding significance.