| Modern food safety includes food hygiene safety and consumer health safety.In recent years,due to unreasonable dietary structure,non-communicable chronic diseases caused by nutritional imbalance have become a social problem that people pay close attention to.The high content of oil in food,especially its unreasonable fatty acid composition,is an important inducement.Chongqing spicy hotpot,a special way of catering in Chongqing,has become popular all over the world.Chongqing spicy hotpot seasoning(Short for hotpot seasoning,the same as below)is made of excessive animal fat(mainly butter),pepper,prickly ash,ginger,garlic,etc.by high-temperature frying and boiling.As a special seasoning food in Chongqing,the hotpot seasoning has been industrially produced and sold at home and abroad.It is not only used in hotpot,but also widely used in family cooking.―Yucai‖ flavor is also derived from the flavor of spicy hotpot.Over the years,there have been health and safety problems in the hotpot seasoning,such as "toxic tripe,industrial pigment,industrial paraffin,hotpot ?Laoyou‘".When eating hotpot,what will happen to the hazard factors(polar components,trans fatty acids)when the hotpot seasoning is heated at high temperature for a long time.In the process of hotpot cooking,vegetables,meat,seafood and so on will be constantly added,which will cause nitrite,nitrate and cholesterol to exceed the standard.The above problems have not been systematically studied so far,as the hotpot seasoning go to the world,the requirements for its quality and safety standards will be more stringent.Hotpot seasoning contains a large number of animal fat,pepper,prickly ash and other spicy irritant substances.Eating hotpot can easily cause gastrointestinal discomfort to consumers,which may lead to gastrointestinal diseases such as proctitis and colitis.How to ensure that there is no mental and physical burden for the diners while enjoying the delicious food,so as to ensure the health and safety of eating hotpot? The research group innovatively combines the safe formula of ingredients and the compound powder of konjac and mulberry leaves(referred to as the compound powder,the same below).It has a good improvement effect on eliminating gastrointestinal discomfort caused by eating hot pot,but its mechanism is not clear.In order to ensure the healthy and sustainable development of Chongqing hotpot industry,this paper comprehensively and systematically studied the health safety and consumption health safety of hotpot seasoning and the mechanism of intervention effect.First of all,the health and safety of hotpot seasoning was evaluated by the detection of hazard factors,exposure risk assessment,repeated heating of hotpot seasoning and the change of hazard factors during cooking dishes for a long time.One hundred groups of hotpot seasoning were collected from Chongqing real estate enterprises and hotpot restaurants.The harmful factors(7 kinds of metals(lead,arsenic,mercury,cadmium,chromium,zinc,manganese,iron,copper),acid value,peroxide value,nitrite,benzo(a)pyrene,polar component,trans fatty acid,seven non edible pigments(basic orange Ⅱ,Rhodamine B,para red,Sudan Ⅰ,Sudan Ⅱ,Sudan Ⅲ,Sudan Ⅳ))and so on)in hotpot seasoning were determined by liquid chromatography,gas chromatography,ion chromatography,atomic absorption spectrophotometry,atomic fluorescence spectrometry and spectrophotometry malondialdehyde.The above risk factors were evaluated by Monte Carlo simulation.The contents of nitrite,nitrate,polar component,trans-fatty acid,cholesterol and other harmful factors in hotpot soup before and after cooking time,repeated cooking times and simulated eating hotpot were determined.Secondly,based on the above health and safety of hotpot seasoning,the mice fed with hotpot seasoning were used as the experimental animal model,and different doses of compound powder were fed to evaluate the health and safety of consumption of hotpot seasoning and the intervention mechanism of compound powder.The levels of TC,TG,HDL-C,LDL-C,AST,ALT,AKP,INF-γ,IL-1β,IL-6,IL-10,IL-4 and TNF-α in serum and liver of mice in different groups were determined by ELISA.He staining method was used to prepare liver and cecum sections.The pathological state of different groups of mice was observed and described by microscope.The cecal contents of different groups of mice were collected under aseptic conditions,and the species and structural diversity of cecal microorganisms were evaluated by 16 SrRNA high-throughput sequencing.Finally,based on PPAR and AMPK/SREBPs signal pathway,RT-qPCR and Western blot were used to verify the molecular mechanism of improving gastrointestinal discomfort caused by chafing dish bottom by regulating intestinal microorganisms.It mainly includes the analysis of gene expression of LPL,CPT1,CYP7A1,PPARα,AMPKα,LXRα,C/EBP α,PPARγ and other mRNA and protein.The research of this paper provides theoretical basis for the modification and improvement of hotpot seasoning standard,provides guarantee for guiding consumers to have better healthy consumption experience for Chongqing spicy hotpot,and is conducive to the effective supervision of regulatory authorities on the health and safety of hotpot seasoning.The main results are as follows:1)Determination of the main components of Chongqing hotpot seasoning and compound powder.The results showed that the contents of water,ash,protein,sodium,carbohydrate and cholesterol were 6.5 g/100 g,10.8 g/100 g,9.4 g/100 g,3.39×103 mg/100 g,9.0 g/100 g and 40 mg/100 g respectively.The fat content was 64.3 g/100 g,1.07 times of the daily nutrient reference value(60g).The total energy is 2691 kJ100 g,of which 2379 kJ/100 g is provided by fat,accounting for 88% of the total energy,exceeding the requirement that the proportion of energy provided by fat in daily diet should not exceed 30% of the total energy.The results showed that the proportion of SFA,UFA and TFA in total fatty acids was 61.847%,35.017% and 3.909% respectively,and the proportion of MUFA and PUFA in total UFA was 31.791% and 3.227% respectively.The content of sodium in the hotpot seasoning is 3.4×103 mg/100 g,which is 1.7 times of the daily nutrient reference value(2.0×103 mg).The content of pepper and hemp in the hotpot seasoning is 457.3 mg/kg,and the content of capsaicin is 129.0 mg/kg.The contents of water,ash,protein,fat,sodium,carbohydrate and energy were 5.8 g/100 g,0.2 g/100 g,0.8 g/100 g,0 g/100 g,0 mg/100 g,57.6 g/100 g and 134.1kJ/100 g respectively.The content of dietary fiber was 35.5 g/100 g,which was 1.42 times higher than the reference value of nutrients(25 g).2)Detection of hazard factors and risk assessment of exposure in hotpot seasoning.First of all,an efficient,rapid and accurate alumina column high pressure liquid chromatography method for the simultaneous determination of 7 kinds of non edible pigments in hotpot seasoning was established.Seven kinds of non edible pigments in hotpot seasoning were extracted by ethanol and n-hexane: acetone(v/v 9:1)mixed solution step by step.After purification by alumina column,the flow of 0.02 mol/L ammonium acetate(pH=4.0)and acetonitrile was carried out by high-pressure liquid chromatography with diode array detector(DAD).The wavelength varied in different periods(0~7min 440nm;7~9min 548nm;9~13min 480nm;13~20min 520nm)Mobile phase,gradient elution,external standard method.Then,the results of hazard factors in 100 groups of hotpot seasoning showed that the unqualified rate of lead,polar component,benzo(a)pyrene was 1%,the unqualified rate of acid value was 18%,arsenic,peroxide value,malondialdehyde,trans fatty acid and 7 kinds of non edible pigments all met the national food safety standards.Finally,based on the data of hazard factors in 100 groups of hotpot seasoning,the risk assessment model of hazard factors exposure was established.The results showed that the risk of eating Chongqing hotpot seasoning to the residents of Chongqing was 0% for arsenic,mercury,copper,zinc,manganese,nitrite,trans fatty acid and 7 kinds of non edible pigments,and the risk of lead,cadmium,chromium,iron,peroxide value and malondialdehyde to the residents of Chongqing was less than 10%.The risk of acid value,polar component and benzo(a)pyrene to Chongqing residents is more than 10%.3)The effects of repeated heating of hotpot seasoning and cooking dishes for a long time on hazard factors were studied.Hotpot seasoning is a complex oil-water mixing system composed of oil,water,powder and solid substances,which will enter into consumers with the hot dishes.The results showed that the contents of nitrate,polar component and cholesterol in hotpot soup increased significantly(p<0.05),the content of nitrite did not change significantly(p>0.05),and the content of trans-fatty acid decreased significantly(p<0.05).With the prolongation of cooking time,the content of nitrite and polar components in hotpot soup increased significantly(p>0.05).4)Effects of compound powder on biochemical indexes,cytokines and histopathological sections of mice fed with chaffy dish bottom material.60 Kunming mice were divided into blank control group(BC),model control group(MC),high dosegroup(HD),medium dosegroup(MD)and low dosegroup(LD)groups.Except BC group,the other groups were given 0.1ml/10 g Chongqing hotpot seasoning(CHS)by gavage.After 30 min,all the mice in the swab group were given compound powder(364,182,91 mg/kg)by gavage for 4W.The results showed that the levels of TC,TG,AST,ALT,AKP,INF-γ,IL-1β,IL-6,IL-10,IL-4 and the ratio of HDL-C to LDL-C in MC group were significantly higher than those in BC group(p<0.05).In MC group,the arrangement of the liver plate was still regular,the balloon like degeneration of the liver cells,a small amount of lymphocyte infiltration,accompanied by focal necrosis of the liver cells,the arrangement of the intestinal glands in the cecum was sparse and irregular,the epithelial cells were exfoliated,the integrity was damaged,the mucosal layer was thinned,the number of goblet cells was significantly reduced,and there was inflammatory cell infiltration.The results showed that the intake of hot hotpot seasoning led to the disorder of lipid metabolism,liver injury and intestinal inflammation in mice.Compared with MC group,TC,TG,LDL-C,AST,ALT,AKP,INF-γ,IL-1β,IL-6,HDL-C,TNF-α,IL-10 and IL-4 in HD group were significantly lower(p<0.05),higher(p<0.05).The morphology of liver and cecum was similar to BC group.It is suggested that the intervention of compound powder can improve the disorder of lipid metabolism,prevent liver injury and intestinal inflammation in mice.5)The effect of compound powder on microbe and short chain fatty acid in cecum of mice fed with hotpot seasoning.The results of 16 SrRNA high-throughput sequencing showed that there were 10 phylum,21 classes,32 orders,62 families and 160 genera of bacteria in the cecum contents of each group of mice.The OTUs determined in the experiment were divided into the following 10 categories: Firmicutes,Bacteroides,Proteobacteria,Actinobacteria,Deferribacteres,Verrucomicrobia,Spirocheaetae,Saccharicteria,Cyanobacteria and Tenericutes.Among them,Firmicutes and Bacteroides belong to dominant flora in all samples,and the sum of their relative abundances is more than 80%.Compared with BC group,MC group had lower OTU number,higher F/B ratio,lower lactobacilli content,higher content of enterobacilli,less microorganism related to carbohydrate transport and metabolism,more microorganism related to fat transport and metabolism.The contents of acetic acid,propionic acid,isobutyric acid,butyric acid,isovaleric acid,valeric acid and total SCFA decreased significantly.The results showed that the intake of hotpot seasoning decreased the species and structural diversity of intestinal microorganisms and the content of SCFA.Compared with MC group,the OTU number of HD group was significantly increased,the cecal microbial diversity was significantly increased,the contents of acetic acid,propionic acid,isobutyric acid,isovaleric acid,valeric acid and total SCFA were significantly increased(p<0.05),the difference of butyric acid content was not significant,but there was still a trend of increase.It indicated that the compound powder could improve the mechanism of fat metabolism disorder and intestinal inflammation in mice by interfering with intestinal microecology.6)The effect of compound powder on the expression of lipid metabolism related genes and proteins in the liver of mice fed with hotpot seasoning.The results showed that the expression levels of LPL,CPT1,CYP7A1,PPAR-α and AMPK-α in MC group were significantly lower than those in BC group(p<0.05),and the expression levels of LXR-α and C/EB-α were significantly higher(p<0.05).The results showed that the expression of genes and proteins related to fat catabolism in the liver of mice decreased,and the expression of genes and proteins related to fat synthesis and metabolism increased.Compared with MC group,the expression levels of LPL,CPT1,CYP7A1,PPAR-α and AMPK-α in HD group were significantly higher(p<0.05),LXR-α and C/EBP-α were significantly lower(p<0.05),PPAR-γ was not significantly higher(p>0.05),but there was a downward trend.The results showed that the compound powder can promote the regulation of cholesterol metabolism,fatty acid β oxidation and fat decomposition in vivo.Further,through the molecular mechanism,it can be verified that the compound powder can improve the disorder of fat metabolism and intestinal inflammation by interfering with intestinal microecology in hotpot seasoning mice. |
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