Investigation Of Volatile Nitrosamines In Animality Preserved Food (Sausage And Salted Fish) On Sale

BackgroundN-nitrosamines compounds are highly damaging of chemical pollutants in the food, which can lead to acute or chronic toxic effect.90% of more than 300 kinds of nitrosamine compounds studied have been proved to induce tumors of different organs of animals,and the volatile nitrosamines toxicity is more outstanding . Food is an important nitrosamine intake source in human. Animality preserved food was deeply loved by the masses of consumers and producers because of its unique flavor and their advantages on the corrosion. In recent years, as living standards improved, people’s attention to food safety has been more and more high, so Animality preserved food security problems have been also in focus. Several researches detected N-nitrosamines compounds in kinds of animal food made in different processing method,and epidemiological studies demonstrated Animality preserved food intake related to risk of gastrointestinal tumor, nasopharyngeal carcinoma.Chinese residents have the habit of eating Animality preserved food, and have a large percentage of sausage and salted fish, so this research collected some sausages and salted fish in our nine province area,and mainly selected sausage and salted fish with high consumption in Guangdong province as the research object, in order to analyze risk of human exposure to volatile nitrosamines in commercially available sausage. This study carried out a cross-sectional investigation of volatile nitrosamines in Animality preserved food sold in high exposed areas in China (sausages and salted fish), monitored the common main content of 9 kinds of volatile nitrosamines, to explore China’s main distribution of volatile nitrosamines high exposed areas, to provide the scientific prof to further animal studies, the crowd and metabolomics research, at the same time to provide direction of making measures and policies to control the main content of volatile nitrosamines in animal preserved food sold.Objective:1. To set up the method of testing 9 kinds of volatile nitrosamines in animal preserved food(sausages and salted fish) at the same time;2. To investigate 9 kinds of volatile nitrosamines, consists of N-nitrosamines (NDMA), N-nitroso diethylamine (NDEA), N-nitroso armour ethylamine (NMEA), N-nitroso propyl amine (NDPA), N-nitroso pyrrolidine (NPYR), N-nitroso piperidine alkanes (NPIP), N-nitroso butyl amine (NDBA), N-nitroso morpholine (NMOR), N-nitroso diphenylamine (NDPHA) in Chinese traditional sausages and salted fish (including ocean fish and freshwater fish)in 9 provinces in China, to compare the detection rate and contents of 9 kinds of volatile nitrosamines in the different regions of Guangdong province, in different production methods (factory production, small workshops or homemade) and in different forms of packaging(vacuum plastic packaging,bulk), etc.3. To explore the concentration of 9 kinds of volatile nitrosamines of sausages and salted fish in other province,and compare contents of 9 kinds of volatile nitrosamines in commercial sausage between other province and guangdong area.4. To analysis the main factors affecting nitrosamine content in sausage and salted fish, in order to take effective preventive measures to reduce nitrosamines content in food, improve the safety of animal preserved food and protect people’s health.Methods1.9 volatile nitrosamines detection method:QuEChERS pretreatment method in combination with gas chromatography-triple quadrupole mass spectrometry.2. Nitrite and nitrate detection method:according to the food and the determination of nitrite and nitrate GB5009.33-2010 standard of the second law: spectrophotometric method determination of nitrate and nitrite content in the samples.3. Statistical method:a database of all data in Excel, using SPSS 20.0 software for statistical analysis. Measurement data expressed with the mean of+/-standard deviation or median(min～max), compared with two independent samples t-test analysis or Mann-Whitney U test analysis, Kruskal Wallis H-inspection analysis used between the different groups, p<0.05 means statistical significance.Results1.Establish a gas chromatography-triple quadrupole mass spectrometry method to detect 9 kinds of volatile nitrosamines in commercial sausage and salted fishThe study used the QuEChERS method and gas chromatography -triple quadrupole mass spectrometry (GC-MS/MS) to separate,purify and test content of N-nitrosodimethylamine(NDMA),N-nitrosodiethylamine(NDEA),N-nitrosomethylet hylamine(NMEA),N-nitrosodipropylamine(NDPA),N-nitrosopyrrolidine(NPYR),N-n itrosopiperidine(NPIP),N-nitrosodibutylamine(NDBA),N-nitrosomorpholine(NMOR ),N-nitrosodiphenylamin(NDPhA) in commercial sausage and salted fish. The linear range of the method is 0.05 ng/mL to 200 ng/mL, detection limit of 0.01～0.1 μg/kg, sausage standard addition recovery is 90.7%～116%, relative standard deviation (RSD) is 1.8%～14.0%; Salted fish standard addition recovery is 77.4%～119.1%, relative standard deviation (RSD) is 0.32%～15.82%. This detection method in the research is of high sensitivity, low detection limit, simple operation and suitable for rapid detection of batch samples.2.Pollution survey of volatile nitrosamines in sell sausages and salted fish from Guangdong provinces and cities(1) Pollution survey of volatile nitrosamines in sell sausages from Guangdong provinces and cities1)This research found that 9 kinds of volatile nitrosamines to some extent detection in six cities and counties in Guangdong province(Zhongshan city, Shantou city, Meizhou city,Shaoguan city, Sihui city) acquisition of 40 cases of commercial sausage samples,NDMA, NDBA, NMOR detection rate of 100%. NDMA content in 15% of the samples (6) is more than the national standards of meat products limited (NDMA 3 μg/kg or less), the highest can amount to 7.37 μg/kg; 20% of the samples (8) of the total content of volatile nitrosamines is more than 10 μg/kg, peak of 15.32 μg/kg.2) No obvious difference among content of total volatile nitrosamines in different commercial sausage sampling area of Guangdong province, but different kinds of volatile nitrosamines varied in different sampling areas, such as the highest content of NDMA in Shantou acquisition of sausages, NMEA sample that is the highest levels of Shaoguan, NDEA higher is the Shantou and Shaoguan, NDPA highest is Zhongshan, NDBA is Guangzhou, the highest NPIP is higher, Meizhou city, NPYR higher is Shaoguan city and Meizhou city, NMOR Sihui is higher.3) Comparing different production mode of commercial sausages in Guangdong areas, NDMA,NDPA,NDBA,NPIP content in the industrialized production of sausage samples are higher than that in small workshops homemade sausage samples; And small workshops in homemade sausage samples NPYR, NMOR, NMEA content are higher than that of industrial production.4) We did not see obvious difference in the content of total volatile nitrosamines in different packing method of sausage sold in Guangdong, but NMOR content of bulk commercial sausage in the Guangdong areas is higher than in the vacuum plastic packaging, the difference was statistically significant.5) The volatile nitrosamines NDMA content is highest in animal liver sausage, then spicy sausage, lowest in ordinary sausage. NDBA is higher in spicy sausage, then animal liver sausage, lowest in ordinary sausage.6) Guangdong province commercial sausage by nitrite, nitrate residue detection results and influence factors analysis of volatile nitrosamines showed, NDMA and is positive relative to nitrite and nitrate residue, NPYR is negative correlation with nitrite and nitrate,NDBA is positive relative to nitrates, NMOR has negative correlation with nitrite. NPYR and nitrite is relatively close, the other does not close correlation (r< 0.5).(2)Pollution survey of volatile nitrosamines in salted fish from Guangdong province1) In total 33 cases of salted fish were collected in Guangdong province and NDMA, NDEA, NDBA, NMOR detection rate was the highest (100%).57.6% of commercially available salted fish samples (19) of NDMA content was more than the limit standard of aquatic products in China (NDMA 4 μg/kg or less), NDMA peak of 88.42μg/kg;total volatile nitrosamines content in 81.8% of commercially available salted fish samples (27) was more than lOug/kg, peak of 104.60μg/kg.2) According to the pollution results of volatile nitrosamines in salted fish of different purchase location,no statistical significance differences was between large supermarkets or brand chains and the farmer’s market or meat market.3) Salted fish is main the bulk on sales approach, in which the sampling results, bulk salted fish in 29 cases, accounting for 88%.4)Three processing methods(Dried,Fried,fermentation) accounted for 97% in different types of salted fish, less smoke processing way.5)The correlation analysis of nitrite (or nitrite)and volatile nitrosamines in commercially salted fish from the Guangdong region, found that the total content of volatile nitrosamines and nitrite residue into negative correlation, NDEA content and nitrate residue into positive correlation, but the relationship is not close (correlation coefficient is less than 0.5).3.Volatile nitrosamines pollution investigation in sold sausages and salted fish from Guangdong provincesIn the commercially available salted fish,for NDMA,NMEA, NDEA,NDBA,NDYR and total volatile nitrosamines,the content is higher than commercial sausage,NDPA and NMOR content in sold sausage is higher than the salted fish from market.4.Nine kinds of volatile nitrosamines content in sold sausages and salted fish from other provinces1) Other provinces sausage were taken from Anhui (10 cases), Guizhou (7 cases), Hunan (14 cases), Shandong (10 cases), Sichuan (13 cases), and NDBA,NMOR,NDMA,NPYR, NDPHA detection rate is higher (> 80%), and NDBA, NMOR detection rate was 100%, the average content were 0.17+/-0.20 μg/kg,0.98+/-1.37μg/kg. NDMA detection rate was 98.1%, and 11.1% of the samples (6) more than the national standards of meat products limited (NDMA 3μg/kg or less), peak of 6.73μg/kg; 35.2% of the samples (19) total volatile nitrosamines is more than lOug/kg, the highest can amount to 100.79μg/kg, which is from hunan homemade sausages, among them NPYR content played the biggest contribution, content of 96. 14μg/kg.2) A total of 21 cases of sold salted fish were collected from other province regions, mainly adopt from the coastal areas of central China,include Zhejiang province(7 cases),Jiangsu province(6 cases),Shandong province (6 cases)and Hunan province (2 cases), For NDMA、NDBA、NMOR,their detection rate is higher (> 80%), and while the NDMA and NDBA detection rate is 100%. NDMA average content is 13.40+/-20.82μg/kg,56% of the samples (14) content is more than the limit standard of aquatic products in China (NDMA 4μg/kg or less), the highest can amount to 89.43 ug/kg;total volatile nitrosamines in 48% of the samples (10)is more than 10μg/kg, content ranging from 13.34 to 93.69 μg/kg.5.The comparison of 9 kinds of volatile nitrosamines in sold sausages between guangdong and other provincesNMEA,NDPA, NPIP content in guangdong sausage is higher than sausages of other provinces, and NDBA, NPYR, NDPHA and total volatile nitrosamines content in other province sausage is higher than guangdong province sausage.6. Comparison of 9 kinds of volatile nitrosamines in seawater and freshwater salted fishThe present study collected 23 cases,7 varieties freshwater fish and 31 cases,9 varieties seawater fish. NDMA and total volatile nitrosamines in seawater salted fish content is higher than that of freshwater salted fish, however, NDPA、NDBA and NMOR content in freshwater salted fish is higher than that in seawater salted fish, and there was no statistically significant difference in other volatile nitrosamines.Conclusion1、A selective gas chromatography tandem triple quadrupole mass spectrometry was used to detected nine volatile nitrosaminesfrom local sausage and salted fish on sale by using QuEChERS sample pretreatment. The developed method was simple, rapid, sensitive and accurate.2、The method was applied in the detection of local sausage and salted fish on sale, and nine volatile nitrosamines were all detected.The detection rate of NDMA、 NDBA、NMOR、NPYR、NDPA、NDPHA is higher.3、The supervision and control for NDMA in sausage and salted fish on sale must word every day,and the pollution of salted fish is more serious.4、the content of total volatile nitrosamines in sausage and salted fish on sale is higher,to refer to the United States department of agriculture (USDA) on total volatile nitrosamines in meat products which cannot exceed 10 ug/kg.which is suggested that our country should establish the standards and testing methods of total volatile nitrosamines in Animality preserved foods to improve the level of food safety and regulation.5、to analysis the influential factors of volatile nitrosamines in sausage,we found the species and contents of nine volatile nitrosamines in local sausage on sale in different regions and different production mode (industrialized production or individual workshop)is different; Bulk of sausages are more easily to pollute by volatile nitrosamines in the process of transporting; For NDBA,the content in sausage is higher by adding spice; The correlation between nitrate or nitrite residue and the content of volatile nitrosamines in local sausage on sale is not close.6、to analysis the influential factors of volatile nitrosamines in salted fish,we found the content from ocean fish is higher from freshwater fish The correlation between nitrate or nitrite residue and the content of volatile nitrosamines in salted fish on sale is not close.the content of volatile nitrosamines of salted fish in different grade Sales site and different packing mode has no statistical significance.it prompt us processing crude and lack of supervision in sale process.