Study On Volatile Evaluation And Tetrodotoxin Detection By In Vivo Sampling In Three Species Of Fugus Of China

Cultured puffer fish have distinctive aroma and desirable taste.There used to be said “Desperate to eat pufferfish”.For a long time,there is a habit of eating puffer fish in China.Recent results will shed a light on the cultured pufferfish production and provide the scientific foundation on the special flavor quality assessment of cultured pufferfish.With the development of economy,the national open of puffer fish edible getting louder and higher,therefore establish a fast,effective and no-injured TTX detection method is security edible puffer fish safety technical support.Based on the three pufferfish: Takifugu obscures,Takifugu flavidusa and Takifugu rubripes as the research object,and then through the comparison of microwave assisted extraction coupling solvent assisted flavor evaporation method?MAE-SAFE?coupling solvent assisted flavor evaporation method and accelerated solvent extraction?ASE-SAFE?to cultured east filefish effect of extracting volatile components,combined with gas chromatography-mass spectrometry?GC-MS?and full two-dimensional gas chromatography/time-of-flight mass spectrometry(GC?GC/TOFMS?combination of volatile components in the qualitative and the quantitative analysis of volatile components in data mining.So as to establish a rapid,accurate and simple method to extract volatile components,for breeding the volatile components of Oriental filefish research foundation,to explore the puffer fish delicious mystery and improving the quality of the puffer fish provides the basic theory of reference data.Established in vivo after dark grain filefish east rapid detection method of tetrodotoxin extraction combining with homemade SPME probe the puffer fugu toxin in vivo for sample preparation method,in order to establish a TTX to track changes in fugu in sample preparation method.The main content of this research and the results are as follows:?1?Accelerated solvent extraction-solvent assisted flavor evaporation?ASE-SAFE?,and microwave-assisted extraction dual solvent assisted flavor evaporation?MAE-SAFE?,were compared for their effectiveness in the extraction of volatile compounds from steamed obscure puffer?Takifugu obscures?meat.The volatile compounds in puffer fish of obscure puffer?Takifugu obscures?,tawny puffer?Takifugu flavidus?and tiger puffer?Takifugu rubripes?extracted by MAE-SAFE?dichloromethane as solvent?were identified using gas chromatography mass spec-trometry?GC-MS?and comprehensive two-dimensional gas chromatography-time of flight mass spectrometry?GC×GC-TOFMS?.Results showed that a total of 215 volatile compounds were identified in 3 kinds of puffer fish.Among them,there were 110 volatile compounds in Takifugu obscures,108 in Takifugu flavidus and 89 in Takifugu rubripes.The extracts obtained from 3 kinds of puffer fish were similar in numbers,but qualitative and quantitative differences of their volatile profiles were observed.Takifugu flavidus,Takifugu obscures and Takifugu rubripes have 30,25 and 13 unique volatile compounds,respectively.?2?Combined with GC-MS analysis showed that fugu and cooked in fugu respectively identified 42 species and 61 kinds of volatile substances,mainly include alcohols,aldehydes,ketones,hydrocarbons,esters,acids,aromatic,nitrogen compounds,phenolic substances,such as only 13 compounds were detected in the two kinds of samples,after cooked the volatile flavor types and content of the puffer fish has changed significantly;Fugu aldehyde content in the highest,including benzene,formaldehyde and 1-octene-3-alcohol content as high as 68.15% and 9.22%;Cooked in fugu alcohols and acids content is higher,the 2,3-butylene glycol?2 r,3r?-?-?-2,3-butanediol content up to 22.80% and 19.84%.In future study,can be in a combination of sensory evaluation and GC-MS analysis,on the basis of electronic nose smell signal data and attempts to establish the puffer fish product quality index of the relational model,and by monitoring the smell changes fast evaluating the freshness of the product,to predict the shelf life of raw fish.Set up electronic nose smell signal data with different processing relationship model of the puffer fish products,odor fingerprint is established.?3?In present study,a rapid and sensitive method based on in vivo solid-phase microextraction?SPME?coupled to liquid chromatography tandem mass spectrometry?LC-MS/MS?was developed to detect the endogenous TTX in pufferfish?Fugu?.Fiber evaluation experiments demonstrated that,compared with the commercial PDMS and PA fibers,the home-made electrospun PS@PDA-GA fibers exhibited much better extraction performance towards to TTX in water?120 times and 20 times,respectively?,under the optimized conditions.Then,the home-made SPME fibers were employed to extract TTX in spiked homogeneous fish muscle samples,and a LC-MS-MS was used for the analysis.The reproducibilities?RSDs of inter and intra fiber were 12.1 %,and 7.9 % respectively?,linear range?1-1000,R2=0.9951?and sensitivity(the LOD was 12.3 ng·g^-1)of the method were found to be excellent and satisfactory for further in vivo experiments.Especially the LOD of the established method is lower than the National Standard Method of China(GB/T 23217-2008,LOD 50 ng·g^-1).Subsequently,the method was successfully applied to detect the TTX in the dorsal muscle of living pufferfish,and the accuracy was verified with traditional liquid extraction?LE?method.In general,this is the first study to detect TTX in pufferfish by in vivo sampling method,which provided a promising alternative method for the studies of TTX,and also advanced the implementation of SPME for more in vivo studies.