CN111398265A - Establishment method of in-vitro nitrosation simulation system for blocking NPYR generation by CSEO proportion in salted meat product - Google Patents
Establishment method of in-vitro nitrosation simulation system for blocking NPYR generation by CSEO proportion in salted meat product Download PDFInfo
- Publication number
- CN111398265A CN111398265A CN202010294693.5A CN202010294693A CN111398265A CN 111398265 A CN111398265 A CN 111398265A CN 202010294693 A CN202010294693 A CN 202010294693A CN 111398265 A CN111398265 A CN 111398265A
- Authority
- CN
- China
- Prior art keywords
- npyr
- nitrosation
- vitro
- test
- essential oil
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 102100031650 C-X-C chemokine receptor type 4 Human genes 0.000 title claims abstract description 97
- 101000922348 Homo sapiens C-X-C chemokine receptor type 4 Proteins 0.000 title claims abstract description 97
- 101000603386 Homo sapiens Neuropeptide Y receptor type 1 Proteins 0.000 title claims abstract description 97
- WNYADZVDBIBLJJ-UHFFFAOYSA-N N-Nitrosopyrrolidine Chemical compound O=NN1CCCC1 WNYADZVDBIBLJJ-UHFFFAOYSA-N 0.000 title claims abstract description 97
- 238000007034 nitrosation reaction Methods 0.000 title claims abstract description 56
- 230000009935 nitrosation Effects 0.000 title claims abstract description 44
- 238000000338 in vitro Methods 0.000 title claims abstract description 42
- 235000013622 meat product Nutrition 0.000 title claims abstract description 41
- 238000004088 simulation Methods 0.000 title claims abstract description 34
- 238000000034 method Methods 0.000 title claims abstract description 33
- 230000000903 blocking effect Effects 0.000 title claims abstract description 24
- 238000012360 testing method Methods 0.000 claims abstract description 106
- 230000005764 inhibitory process Effects 0.000 claims abstract description 95
- 230000004044 response Effects 0.000 claims abstract description 53
- UMFJAHHVKNCGLG-UHFFFAOYSA-N n-Nitrosodimethylamine Chemical compound CN(C)N=O UMFJAHHVKNCGLG-UHFFFAOYSA-N 0.000 claims abstract description 47
- 230000000694 effects Effects 0.000 claims abstract description 25
- 239000003112 inhibitor Substances 0.000 claims abstract description 21
- 238000012545 processing Methods 0.000 claims abstract description 15
- 238000012795 verification Methods 0.000 claims abstract description 15
- 238000004364 calculation method Methods 0.000 claims abstract description 4
- 239000000341 volatile oil Substances 0.000 claims description 120
- 238000013461 design Methods 0.000 claims description 29
- 235000016639 Syzygium aromaticum Nutrition 0.000 claims description 25
- 244000223014 Syzygium aromaticum Species 0.000 claims description 25
- 241000949456 Zanthoxylum Species 0.000 claims description 24
- 240000004760 Pimpinella anisum Species 0.000 claims description 23
- 235000012550 Pimpinella anisum Nutrition 0.000 claims description 23
- 244000178231 Rosmarinus officinalis Species 0.000 claims description 21
- 235000015241 bacon Nutrition 0.000 claims description 19
- 238000006243 chemical reaction Methods 0.000 claims description 15
- 239000000839 emulsion Substances 0.000 claims description 14
- 239000007853 buffer solution Substances 0.000 claims description 13
- 239000000243 solution Substances 0.000 claims description 12
- 235000007265 Myrrhis odorata Nutrition 0.000 claims description 10
- 230000003993 interaction Effects 0.000 claims description 10
- 240000004160 Capsicum annuum Species 0.000 claims description 9
- 235000008534 Capsicum annuum var annuum Nutrition 0.000 claims description 9
- 238000002835 absorbance Methods 0.000 claims description 9
- RUFPHBVGCFYCNW-UHFFFAOYSA-N 1-naphthylamine Chemical compound C1=CC=C2C(N)=CC=CC2=C1 RUFPHBVGCFYCNW-UHFFFAOYSA-N 0.000 claims description 8
- HVBSAKJJOYLTQU-UHFFFAOYSA-N 4-aminobenzenesulfonic acid Chemical compound NC1=CC=C(S(O)(=O)=O)C=C1 HVBSAKJJOYLTQU-UHFFFAOYSA-N 0.000 claims description 8
- CDBYLPFSWZWCQE-UHFFFAOYSA-L Sodium Carbonate Chemical compound [Na+].[Na+].[O-]C([O-])=O CDBYLPFSWZWCQE-UHFFFAOYSA-L 0.000 claims description 8
- LPXPTNMVRIOKMN-UHFFFAOYSA-M sodium nitrite Chemical compound [Na+].[O-]N=O LPXPTNMVRIOKMN-UHFFFAOYSA-M 0.000 claims description 8
- AKYUZBFQLWFNOB-UHFFFAOYSA-K trisodium 2-hydroxypropane-1,2,3-tricarboxylate hydrochloride Chemical compound [Na+].[Na+].[Na+].Cl.[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O AKYUZBFQLWFNOB-UHFFFAOYSA-K 0.000 claims description 8
- 239000011521 glass Substances 0.000 claims description 7
- 238000002474 experimental method Methods 0.000 claims description 6
- 244000089698 Zanthoxylum simulans Species 0.000 claims description 5
- 238000004945 emulsification Methods 0.000 claims description 5
- 238000010200 validation analysis Methods 0.000 claims description 5
- 229920002594 Polyethylene Glycol 8000 Polymers 0.000 claims description 4
- 238000011161 development Methods 0.000 claims description 4
- 230000001804 emulsifying effect Effects 0.000 claims description 4
- 238000013433 optimization analysis Methods 0.000 claims description 4
- 229910000029 sodium carbonate Inorganic materials 0.000 claims description 4
- 235000010288 sodium nitrite Nutrition 0.000 claims description 4
- 229950000244 sulfanilic acid Drugs 0.000 claims description 4
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 4
- FCLZCOCSZQNREK-UHFFFAOYSA-N Pyrrolidine, hydrochloride Chemical compound Cl.C1CCNC1 FCLZCOCSZQNREK-UHFFFAOYSA-N 0.000 claims description 3
- 238000007405 data analysis Methods 0.000 claims description 3
- 239000007788 liquid Substances 0.000 claims description 3
- 238000013507 mapping Methods 0.000 claims description 3
- 230000001678 irradiating effect Effects 0.000 claims description 2
- 238000007781 pre-processing Methods 0.000 claims description 2
- XKLJHFLUAHKGGU-UHFFFAOYSA-N nitrous amide Chemical compound ON=N XKLJHFLUAHKGGU-UHFFFAOYSA-N 0.000 abstract description 12
- 238000005259 measurement Methods 0.000 abstract 1
- 235000008184 Piper nigrum Nutrition 0.000 description 21
- 241000722363 Piper Species 0.000 description 17
- 235000002566 Capsicum Nutrition 0.000 description 14
- 239000006002 Pepper Substances 0.000 description 14
- 235000016761 Piper aduncum Nutrition 0.000 description 14
- 235000017804 Piper guineense Nutrition 0.000 description 14
- 235000013305 food Nutrition 0.000 description 12
- 239000000796 flavoring agent Substances 0.000 description 11
- 235000019634 flavors Nutrition 0.000 description 11
- 244000061520 Angelica archangelica Species 0.000 description 9
- 235000001287 Guettarda speciosa Nutrition 0.000 description 9
- 230000002829 reductive effect Effects 0.000 description 9
- 235000013599 spices Nutrition 0.000 description 9
- 230000015572 biosynthetic process Effects 0.000 description 8
- 244000203593 Piper nigrum Species 0.000 description 7
- 238000004458 analytical method Methods 0.000 description 7
- 235000013614 black pepper Nutrition 0.000 description 7
- 239000003921 oil Substances 0.000 description 7
- 244000131415 Zanthoxylum piperitum Species 0.000 description 6
- 235000008853 Zanthoxylum piperitum Nutrition 0.000 description 6
- 235000013372 meat Nutrition 0.000 description 6
- 239000010649 ginger oil Substances 0.000 description 5
- 241001127714 Amomum Species 0.000 description 4
- 241000234314 Zingiber Species 0.000 description 4
- 235000006886 Zingiber officinale Nutrition 0.000 description 4
- 239000002131 composite material Substances 0.000 description 4
- 235000013399 edible fruits Nutrition 0.000 description 4
- 235000008397 ginger Nutrition 0.000 description 4
- 238000004519 manufacturing process Methods 0.000 description 4
- 238000011160 research Methods 0.000 description 4
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 3
- 238000000605 extraction Methods 0.000 description 3
- 239000000203 mixture Substances 0.000 description 3
- 230000001105 regulatory effect Effects 0.000 description 3
- 235000007650 Aralia spinosa Nutrition 0.000 description 2
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 2
- YGJHZCLPZAZIHH-UHFFFAOYSA-N N-Nitrosodi-n-butylamine Chemical compound CCCCN(N=O)CCCC YGJHZCLPZAZIHH-UHFFFAOYSA-N 0.000 description 2
- WBNQDOYYEUMPFS-UHFFFAOYSA-N N-nitrosodiethylamine Chemical compound CCN(CC)N=O WBNQDOYYEUMPFS-UHFFFAOYSA-N 0.000 description 2
- IOVCWXUNBOPUCH-UHFFFAOYSA-M Nitrite anion Chemical compound [O-]N=O IOVCWXUNBOPUCH-UHFFFAOYSA-M 0.000 description 2
- 238000000540 analysis of variance Methods 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 239000010634 clove oil Substances 0.000 description 2
- 230000003247 decreasing effect Effects 0.000 description 2
- 238000010586 diagram Methods 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 239000003344 environmental pollutant Substances 0.000 description 2
- 235000019441 ethanol Nutrition 0.000 description 2
- 235000011389 fruit/vegetable juice Nutrition 0.000 description 2
- 230000002401 inhibitory effect Effects 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 238000005457 optimization Methods 0.000 description 2
- 231100000719 pollutant Toxicity 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 239000010668 rosemary oil Substances 0.000 description 2
- 229940058206 rosemary oil Drugs 0.000 description 2
- 231100000419 toxicity Toxicity 0.000 description 2
- 230000001988 toxicity Effects 0.000 description 2
- 244000141331 Amomum villosum Species 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- 101000603420 Homo sapiens Nuclear pore complex-interacting protein family member A1 Proteins 0.000 description 1
- YLKFDHTUAUWZPQ-UHFFFAOYSA-N N-Nitrosodi-n-propylamine Chemical compound CCCN(N=O)CCC YLKFDHTUAUWZPQ-UHFFFAOYSA-N 0.000 description 1
- UWSDONTXWQOZFN-UHFFFAOYSA-N N-nitrosopiperidine Chemical compound O=NN1CCCCC1 UWSDONTXWQOZFN-UHFFFAOYSA-N 0.000 description 1
- 102100038845 Nuclear pore complex-interacting protein family member A1 Human genes 0.000 description 1
- 244000269722 Thea sinensis Species 0.000 description 1
- YUWBVKYVJWNVLE-UHFFFAOYSA-N [N].[P] Chemical compound [N].[P] YUWBVKYVJWNVLE-UHFFFAOYSA-N 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- -1 amine compound Chemical class 0.000 description 1
- 230000003064 anti-oxidating effect Effects 0.000 description 1
- 230000003260 anti-sepsis Effects 0.000 description 1
- 239000003963 antioxidant agent Substances 0.000 description 1
- 230000003078 antioxidant effect Effects 0.000 description 1
- 230000033228 biological regulation Effects 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- 230000000711 cancerogenic effect Effects 0.000 description 1
- 231100000315 carcinogenic Toxicity 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
- 238000000354 decomposition reaction Methods 0.000 description 1
- 230000007423 decrease Effects 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 230000006866 deterioration Effects 0.000 description 1
- XHFGWHUWQXTGAT-UHFFFAOYSA-N dimethylamine hydrochloride Natural products CNC(C)C XHFGWHUWQXTGAT-UHFFFAOYSA-N 0.000 description 1
- IQDGSYLLQPDQDV-UHFFFAOYSA-N dimethylazanium;chloride Chemical compound Cl.CNC IQDGSYLLQPDQDV-UHFFFAOYSA-N 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 238000005485 electric heating Methods 0.000 description 1
- 239000003995 emulsifying agent Substances 0.000 description 1
- LIWAQLJGPBVORC-UHFFFAOYSA-N ethylmethylamine Chemical compound CCNC LIWAQLJGPBVORC-UHFFFAOYSA-N 0.000 description 1
- 238000003050 experimental design method Methods 0.000 description 1
- 235000013373 food additive Nutrition 0.000 description 1
- 239000002778 food additive Substances 0.000 description 1
- 239000007789 gas Substances 0.000 description 1
- 239000000383 hazardous chemical Substances 0.000 description 1
- 239000010460 hemp oil Substances 0.000 description 1
- 230000002452 interceptive effect Effects 0.000 description 1
- 150000002576 ketones Chemical class 0.000 description 1
- 230000000670 limiting effect Effects 0.000 description 1
- 238000012417 linear regression Methods 0.000 description 1
- 150000002632 lipids Chemical class 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 231100000243 mutagenic effect Toxicity 0.000 description 1
- 230000003505 mutagenic effect Effects 0.000 description 1
- IRJPMACBKNQUFU-UHFFFAOYSA-N n-(nitrosomethyl)ethanamine Chemical compound CCNCN=O IRJPMACBKNQUFU-UHFFFAOYSA-N 0.000 description 1
- 150000002826 nitrites Chemical class 0.000 description 1
- 230000003647 oxidation Effects 0.000 description 1
- 238000007254 oxidation reaction Methods 0.000 description 1
- 150000002989 phenols Chemical class 0.000 description 1
- 150000008442 polyphenolic compounds Chemical class 0.000 description 1
- 235000013824 polyphenols Nutrition 0.000 description 1
- 230000008092 positive effect Effects 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 230000000979 retarding effect Effects 0.000 description 1
- 239000001509 sodium citrate Substances 0.000 description 1
- NLJMYIDDQXHKNR-UHFFFAOYSA-K sodium citrate Chemical compound O.O.[Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O NLJMYIDDQXHKNR-UHFFFAOYSA-K 0.000 description 1
- 238000007619 statistical method Methods 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 238000000194 supercritical-fluid extraction Methods 0.000 description 1
- 230000001629 suppression Effects 0.000 description 1
- 238000005211 surface analysis Methods 0.000 description 1
- 231100000378 teratogenic Toxicity 0.000 description 1
- 230000003390 teratogenic effect Effects 0.000 description 1
- 238000010998 test method Methods 0.000 description 1
- 238000000870 ultraviolet spectroscopy Methods 0.000 description 1
Images
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/75—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
- G01N21/77—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator
- G01N21/78—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator producing a change of colour
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
- G01N21/31—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry
- G01N21/33—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry using ultraviolet light
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/75—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
- G01N21/77—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator
- G01N21/78—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator producing a change of colour
- G01N2021/786—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator producing a change of colour with auxiliary heating for reaction
Landscapes
- Physics & Mathematics (AREA)
- Chemical & Material Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Spectroscopy & Molecular Physics (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Plasma & Fusion (AREA)
- Engineering & Computer Science (AREA)
- Meat, Egg Or Seafood Products (AREA)
Abstract
The invention relates to a method for establishing an in-vitro nitrosation simulation system for generating a CSEO ratio blocking NPYR in a salted meat product, which comprises ⑴ in-vitro nitrosation reaction simulation, ⑵ NPYR measurement, ⑶ single-factor test, ⑷ response surface center combination test, ⑸ NPYR and NDMA inhibition rate calculation, ⑹ data processing and ⑹ data processing, wherein the model preferably obtains an in-vitro nitrosation simulation system establishment method for generating the CSEO ratio blocking NPYR in the salted meat product, the prediction value of the NPYR inhibition rate is 69.84%, the inhibition rate obtained by the verification test is (68.40% +/-0.33%), the inhibition rate of the ratio on NDMA is (66.69 +/-0.43%), the inhibition rates of the ratio on the NDMA are respectively 66.13% and 65.99%, and the obtained in-vitro nitrosation simulation system establishment method has good actual effect and practical value, and provides new theoretical data for selecting and supporting the meat product according to the industrial N-nitrosamine inhibitor.
Description
Technical Field
The invention belongs to the field of food processing, relates to a food safety technology, and particularly relates to a method for establishing an in-vitro nitrosation simulation system for blocking NPYR generation by CSEO proportion in a salted meat product.
Background
In recent years, with the rapid increase of economic strength of China, the safety problem of the food processing industry is more and more emphasized. In the field of meat product processing, nitrites are widely used as food additives because they can impart a stable red color to meat products, inhibit the growth and propagation of harmful bacteria, and stabilize the oxidation of lipids in the products during storage, thereby retarding the deterioration of food products and improving the flavor of meat. However, nitrite is very easy to have nitrosation reaction with amine compound, which is a protein decomposition product rich in the salted meat product, under the weak acid condition, and finally a hazardous substance (N-nitrosamine) with carcinogenic, teratogenic and mutagenic properties is generated. Thus, in the processing of salted meat products, the production of N-nitrosamines is often reduced or inhibited by the addition of nitrosation inhibitors.
At present, researches on how to control the formation of N-nitrosamine at home and abroad mainly focus on finding natural nitrosation inhibitors such as VC, VE, tea polyphenol, Spice Essential Oil (SEO) and the like, wherein the SEO not only has unique smell and can correct and color, but also has the effects of bacteriostasis, antisepsis, antioxidation and the like. The mechanism of SEO for blocking N-nitrosamine is to utilize its antioxidant component (such as phenols and ketones) to denitrify the nitrosation reagent N2O3Reduction to N2NO, and the like, thereby effectively eliminating the residual quantity of nitrite and blocking the generation of N-nitrosamine.
N-dimethylnitrosamine (NDMA) is one of N-nitrosamine with strong toxicity, and China stipulates the NDMA limit in GB 2762-2017 pollutant limit in food safety standard food, meat and meat products (except meat cans) have the content of not more than 3.0 mu g/kg, and through the design of a secondary regression orthogonal combination test, in an in vitro simulation nitrosation reaction system, Composite Spice Essential Oil (CSEO) for blocking the generation of the NDMA is preferably obtained, wherein the composite spice essential oil comprises 2.078m black pepper L/L, ginger juice 3.118m L/L, 3.692m L/L, 2.833m L/L and clove 0.218m L/L, the suppression rate of the NDMA reaches 64.41%, and when the CSEO is applied to the processing of the Western-type bacon, the NDMA content can be remarkably reduced, while the NDMA content in the group of the N-Nitrosoamine (NPMA) is reduced by 56.47 g-47%, and the NDMA content in other groups is also reduced by 56.51.47% or 65%.
Although the content of NPYR in meat and meat products is not regulated by corresponding standards promulgated by China, the NPYR contained in meat products sold in the American market is regulated by the U.S. food and drug administration to be not more than 10 mug/kg. Therefore, in view of ensuring the safety quality of the western bacon, the present invention screens CSEO that can effectively block NPYR with the goal of blocking NPYR formation as an assessment.
The response surface design method is a statistical method which utilizes a reasonable experimental design method and obtains certain data through experiments, adopts a multivariate quadratic regression equation to fit the functional relationship between factors and response values, seeks optimal process parameters through the analysis of the regression equation and solves the multivariate problem. The invention adopts a method of combining single factor and center combination test design of response surface, firstly selects 5 SEOs (pepper essential oil, anise essential oil, pepper essential oil, clove essential oil, rosemary essential oil, black pepper essential oil, angelica flavor ginger and amomum fruit essential oil) with better NPYR inhibition rate from 8 kinds of commercial SEOs, and then screens the optimal proportion of the 5 SEOs by the center combination test design of the response surface. Finally, the optimized CSEO is subjected to a verification test, the inhibition effect on NPYR is examined, and the inhibition rate on NDMA is also made, so that the purpose of laying a foundation for the future application of the CSEO in salted meat products such as western bacon and the like is achieved.
Disclosure of Invention
The invention aims to overcome the defects of the prior art, and provides a method for establishing an in-vitro nitrosation simulation system generated by blocking NPYR by a CSEO ratio in a salted meat product, wherein the predicted value of the NPYR inhibition ratio obtained by model optimization is 69.84%, the inhibition ratio obtained by a verification test is (68.40 +/-0.33)%, and the inhibition ratio of the ratio to NDMA is (66.69 +/-0.43)%, the inhibition ratios of the NPYR and the NDMA are 66.13% and 65.99% respectively, and the obtained regression model has good fitting effect with the reality and practical value, and provides a new theoretical basis and data support for the selection of the N-nitrosamine inhibitor in the meat product industry.
The technical problem to be solved by the invention is realized by adopting the following technical scheme:
an in-vitro nitrosation simulation system establishment method for blocking NPYR generation by CSEO proportion in salted meat products comprises the following specific steps:
⑴ in vitro simulation of nitrosation reaction
a, respectively sucking SEO with different volumes into a 50m L centrifugal tube, adding 0.05g PEG8000, and then adding 0.5 mol/L sodium citrate-hydrochloric acid buffer solution to make the total volume of the SEO to be 10.0m L;
b, homogenizing and emulsifying the solution obtained in the step a for 1min, putting 1.0m L prepared emulsion into a 50m L centrifuge tube, sequentially adding 0.5 mol/L sodium citrate-hydrochloric acid buffer solution 5.0m L, 1 mmol/L sodium nitrite 0.5m L, 1 mmol/L pyrrolidine hydrochloride 0.5m L and 3m L buffer solution to enable the total volume of the reaction system to be 10.0m L, and carrying out water bath reaction for 1h at 37 ℃;
⑵ NPYR determination
a, after the in vitro simulation nitrosation reaction in the step ⑴ is finished, taking 1.0m L reaction solution into a glass test tube, adding 0.5% sodium carbonate solution 0.5m L to terminate the reaction, and placing the glass test tube under a 15W ultraviolet lamp in a super clean bench for irradiating for 15 min;
b, taking out, immediately and sequentially adding 1% of sulfanilic acid 1.5m L, 0.1% of α -naphthylamine 1.5m L and 0.5m L buffer solution until the volume of the solution is accurate to 5.0m L, and shaking up;
c, standing for 15min for color development, and then carrying out colorimetric determination on the absorbance at the wavelength of 525 nm;
⑶ one-factor test
a, sucking 0.00, 0.05, 0.50, 1.00, 1.50 and 2.00m L for each SEO respectively to ensure that the final concentration of each SEO in the in-vitro simulated nitrosation reaction is 0.0, 0.5, 5.0, 10.0, 15.0 and 20.0m L/L respectively;
b, after the reaction is finished, carrying out NPYR determination, wherein a blank control group is blank emulsion without SEO;
c, calculating the inhibition rate of each SEO on NPYR at the corresponding volume concentration, and repeating the test for 3 times;
⑷ center of response surface combination test
On the basis of a single-factor test result, the SEO volume concentration is screened, the SEO volume concentration is taken as an influence factor level, the NPYR inhibition rate is taken as a response value, Design-Expert7.0 software is used, and a response surface optimization analysis test is designed according to a Box-Behnken center combined test Design principle;
⑷ validation test
Verification in vitro nitrosation simulation emulsification system
Adding the CSEO proportion obtained according to the design of the response surface center combined test into a system formed by in vitro simulation NPYR, and performing 6 times of repeated verification tests;
⑸ calculation of NPYR and NDMA inhibition rates
(A0Is the absorbance value of the blank emulsion without SEO; a. thexAs absorbance value upon addition of SEO emulsion).
⑹ data processing.
Further, in step ⑴, the pH of the sodium citrate-hydrochloric acid buffer is 3.0.
In addition, in the step ⑴, a homogenizer is adopted for homogenizing and emulsifying, and the B gear rotating speed is adjusted to 13000 r/min.
Furthermore, the distance between the liquid level of the tube and the tube is 15cm in step ⑵.
And the CSEO proportion obtained by the response surface center combination design is applied to the western-style bacon, the inhibition effect of the addition of the nitrosation inhibitor on NPYR and NDMA in the western-style bacon is analyzed, meanwhile, a control experiment is carried out, and the test result of the combination design is further verified.
In addition, in the single-factor test, the SEO is 8 types, namely zanthoxylum essential oil, aniseed essential oil, pimento essential oil, clove essential oil, rosemary essential oil, black pepper essential oil, angelica flavor ginger oil and amomum fruit essential oil, the volume concentration of the rosemary essential oil is 20.0m L/L, the zanthoxylum essential oil is 5.0-10.0 m L0/L1, the pimento essential oil is 0.5m L/L, the clove essential oil is 15.0m L/L, the angelica flavor ginger oil is 10.0m L/L, and the aniseed essential oil is 15.0m L/L.
And analyzing the inhibition effect of the nitrosation inhibitor on NPYR and NDMA by taking the volume concentration of pepper essential oil (A, 330.0-470.0 mu L/L), the volume concentration of anise essential oil (B, 300.0-370.0 mu L/L), the volume concentration of pepper essential oil (C, 13.0-20.0 mu L/L), the volume concentration of clove essential oil (D, 10.0-17.0 mu L/L) and the volume concentration of rosemary essential oil (E, 100.0-170.0 mu L/L) as influence factor levels and the NPYR inhibition rate as a response value, wherein the code of the factor levels is as follows:
and the CSEO proportion obtained by the response surface center combination design is applied to the western-style bacon, the inhibition effect of the addition of the nitrosation inhibitor on NPYR and NDMA in the western-style bacon is analyzed, meanwhile, a control experiment is carried out, and the test result of the combination design is further verified.
Moreover, the volume concentrations of the zanthoxylum essential oil, the aniseed essential oil, the pimento essential oil, the clove essential oil and the rosemary essential oil are 441.0, 337.0, 17.0, 14.0 and 137.0 mu L/L respectively, the predicted value of the NPYR inhibition rate is 69.84 percent according to a model, the inhibition rate obtained by a verification test is (68.40 +/-0.33 percent), the inhibition rate of the mixture ratio on NDMA is (66.69 +/-0.43 percent), the inhibitor at the level is added into a meat product (Western bacon), the inhibition rates of the NPYR and the NDMA are 66.13 percent and 65.99 percent respectively,
furthermore, the data processing was performed by preprocessing the experimental data with Microsoft Excel 2010, data analysis with statistix8.1, significant differences (P <0.05) by Turkey test program, Sigmaplot10.0 mapping.
Moreover, Design-Expert7.0 Design is adopted in response surface test, result analysis is carried out, and interaction effects of any two factors are analyzed and evaluated by establishing a regression equation, drawing contour lines and a response surface diagram.
The invention has the advantages and positive effects that:
1. in the method, NDMA is one of N-nitrosamine with strong toxicity, NDMA limit is regulated in GB 2762-2017 pollutant limit in food safety standard food, and the content of meat and meat products (except meat cans) is not more than 3.0 mu g/kg in China, the inventor designs an alcohol extraction Composite Spice Essential Oil (CSEO) for blocking the generation of NDMA in an in vitro simulation nitrosation reaction system through a secondary regression orthogonal combination test in the earlier stage, wherein the proportion of the alcohol extraction composite spice essential oil is black pepper 2.078m L/L, ginger juice 3.118m L/L, pepper 3.692m L/L, 2.833m L/L and clove 0.218m L/L, the inhibition rate of NDMA reaches 64.41%, when CSEO is applied to the processing of the western-type roots, the NDMA content can be remarkably reduced (the NDMA content is 0.218m L/L), the inhibition rate of NDMA is reduced by the NPR limit of the NPR 65), and the other products are not subjected to the NPR inhibition rate regulation of the NPR 65, and the NPR limit is reduced by the NPR limit of the NPR 65, and the NPR limit of the national standard food safety standard food products (the NPR limit), and the NPR limit is not reduced by the NPR 65, the NPR limit of the U.S, the national standard food safety standard food products are considered in the U.
2. The invention adopts a method of combining single factor and center combination test design of response surface, firstly selects 5 SEOs (pepper essential oil, anise essential oil, pepper essential oil, clove essential oil, rosemary essential oil, black pepper essential oil, angelica flavor ginger and amomum fruit essential oil) with better NPYR inhibition rate from 8 kinds of commercial SEOs, and then screens the optimal proportion of the 5 SEOs by the center combination test design of the response surface. Finally, the optimized CSEO is subjected to a verification test, the inhibition effect on NPYR is examined, and the inhibition rate on NDMA is also made, so that the purpose of laying a foundation for the future application of the CSEO in salted meat products such as western bacon and the like is achieved.
3. The invention provides an in-vitro nitrosation simulation system establishment method for blocking NPYR generation by CSEO proportion in salted meat products through single-factor tests and response surface center combination tests, which is verified by western bacon, the model preferably obtains that the predicted value of NPYR inhibition rate is 69.84%, the verification test obtains that the inhibition rate is (68.40 +/-0.33)%, the inhibition rate of the proportion on NDMA is (66.69 +/-0.43)%, the in-vitro nitrosation simulation system establishment method for blocking NPYR generation by CSEO proportion in salted meat products is established, the inhibitor of the level is added into meat products (western bacon), the inhibition rates of NPYR and NDMA are 66.13% and 65.99%, the obtained regression model has good fitting effect with practical value, and new theoretical basis and data support are provided for the selection of N-nitrosamine inhibitors in the meat product industry.
Drawings
FIG. 1 is a graph of the response of prickly ash and pricklyash peel of the present invention to inhibition;
FIG. 2 is a contour plot of inhibition of Zanthoxylum bungeanum and Zanthoxylum bungeanum according to the present invention;
FIG. 3 is a graph of the response of anise and clove to inhibition according to the present invention;
FIG. 4 is a line contour plot of the inhibition of anise and clove of the present invention.
Detailed Description
The present invention is further illustrated by the following specific examples, which are intended to be illustrative, not limiting and are not intended to limit the scope of the invention.
1 materials and methods
1.1 test materials
Commercially available 8 types of supercritical extraction SEO (zanthoxylum essential oil, anise essential oil, pimento essential oil, clove essential oil, rosemary essential oil, black pepper essential oil, angelica flavor ginger oil, amomum fruit essential oil), Tianjin, spring science and technology development Co., Ltd, emulsifier (PEG8000), sodium citrate, hydrochloric acid, α -naphthylamine, absolute ethyl alcohol, anhydrous sulfanilic acid, sodium nitrite, dimethylamine hydrochloride, sodium carbonate, all analytically pure, national medicine group chemical agents Co., Ltd, 7 types of N-nitrosamine standards, NDMA, N-diethylnitrosamine (NDEA), N-methylethylnitrosamine (N-nitrosylmethylethylamine, NMEA), N-dibutylnitrosamine (N-nitrosodibutylnitrosamine, NDBA), N-dipropylnitrosamine (N-nitrosodipropylnitrosamine, NDPA), NPIP, NPYR, Sigma USA.
STARTER3100 acidimeter, Ohaus, USA; 18Basic homogenizer, IKA corporation, germany; XMTD-4000 electric heating constant temperature water bath, Shanghai Ke Heng industrials development Limited company; NU-425-; t6 New century type ultraviolet-visible spectrophotometer, Beijing Pujingyao general instruments, Inc.; 7890A gas chromatograph equipped with a nitrogen phosphorus detector, Agilent, USA.
1.2 test methods
1.2.1 preparation of in vitro simulation nitrosation emulsification System
Respectively sucking SEO with different volumes to 50m L centrifuge tubes with screw caps, adding 0.05g PEG8000, adding a certain volume of 0.5 mol/L sodium citrate-hydrochloric acid buffer solution (pH 3.0) to make the total volume of 10.0m L0, emulsifying for 1min by using a homogenizer B baffle (13000r/min), taking 1.0m L1 prepared emulsion to 50m L2 centrifuge tubes with screw caps, sequentially adding 0.5 mol/L sodium citrate-hydrochloric acid buffer solution (pH 3.0)5.0m L, 1 mmol/L sodium nitrite 0.5m L, 1 mmol/L pyrrolidine hydrochloride 0.5m L and 3m L buffer solution to make the total volume of the reaction system 10.0m L, and carrying out water bath reaction for 1h at 37 ℃.
1.2.2 method for measuring NPYR
After the in vitro simulation nitrosation reaction is finished according to 1.2.1, taking 1.0m L reaction solution into a glass test tube, adding 0.5% sodium carbonate solution 0.5m L to terminate the reaction, placing the glass test tube under a 15W ultraviolet lamp in a clean bench for 15min (the distance between the liquid level of the test tube and a lamp tube is 15cm), taking out the glass test tube, sequentially adding 1% sulfanilic acid 1.5m L (shaking up), 0.1% α -naphthylamine 1.5m L (shaking up) and 0.5m L buffer solution until the volume of the solution is accurate to 5.0m L, shaking up, standing for 15min for color development, and carrying out colorimetric determination on the solution at the wavelength of 525nm to determine the absorbance.
1.2.3 Single factor test
The inhibition rate of 8 SEO emulsions with different volume concentrations on NPYR is studied, 8 SEO emulsions with different volume concentrations are respectively absorbed by 0.00, 0.05, 0.50, 1.00, 1.50 and 2.00m L according to a method of 1.2.1, so that the final concentration of each SEO in the in-vitro simulated nitrosation reaction is respectively 0.0, 0.5, 5.0, 10.0, 15.0 and 20.0m L/L, after the reaction is finished, the NPYR is measured according to 1.2.2, a blank control group is a blank emulsion without adding spice essential oil, the inhibition rate of each SEO on the NPYR at the corresponding volume concentration is calculated, and the test is repeated for 3 times.
1.2.4 response surface center combination test design
On the basis of the single-factor test result, the volume concentration (A, 330.0-470.0 mu L/L) of the zanthoxylum essential oil, the volume concentration (B, 300.0-370.0 mu L/L) of the aniseed essential oil, the volume concentration (C, 13.0-20.0 mu L/L) of the zanthoxylum essential oil, the volume concentration (D, 10.0-17.0 mu L/L) of the clove essential oil and the volume concentration (E, 100.0-170.0 mu L/L) of the rosemary essential oil are finally used as influencing factor levels, the NPYR inhibition rate is used as a response value, a Design-Expert7.0 software is used, a response surface optimization analysis test is designed according to a Box-Behnken center combined test Design principle, and an encoding factor level table is shown in Table 1.
TABLE 1 response surface test factors and horizontal coding values
1.2.5 validation test
1.2.5.1 validation in vitro nitrosation simulated emulsification System
The CSEO optimal proportion designed by the response surface center combination test is added into a system formed by in vitro simulation NPYR according to 1.2.4, and 6 times of repeated verification tests are carried out. To understand how the preferred CSEO inhibits NDMA formation in vitro, the rate of inhibition of NDMA formation by CSEO in a simulated system was also determined.
1.2.5.2 validation in Western bacon
The optimum proportioning nitrosation inhibitor obtained by the combined design of the center of the response surface is applied to the actual production of the western-style bacon, the inhibition effect of the addition of the nitrosation inhibitor on NPYR and NDMA in the western-style bacon is analyzed, meanwhile, a control experiment is carried out (the addition proportion of the rest raw materials is consistent with that of a test group except that the nitrosation inhibitor is not added in the control group of the western-style bacon), and the test result of the combined design is further verified.
1.2.6 calculation of NPYR and NDMA inhibition
(A0Is the absorbance value of the blank emulsion without SEO; a. thexAs absorbance value upon addition of SEO emulsion).
1.3 data processing
The experimental data were pre-processed with Microsoft Excel 2010, data analysis was performed with Statistix8.1 and significant differences (P <0.05) were performed by Turkey test program, Sigmaplot10.0 mapping. The response surface test is designed by adopting Design-Expert7.0 and results are analyzed, and the interaction effect of any two factors is analyzed and evaluated by establishing a regression equation, drawing contour lines and a response surface graph.
2 results and analysis
2.1 Single factor test
TABLE 28 inhibition of NPYR (%). by the addition of different concentrations of spice essential oils
Note: the capital letters in different shoulders indicate that the difference between different volume concentrations of the same spice essential oil is significant (P < 0.05).
The inhibition rate of 8 SEO to NPYR changes in different addition concentrations is shown in Table 2, it is known from Table 2 that the inhibition rate of 7 SEO to NPYR is strongest, the inhibition effect is also obviously enhanced with the increase of the volume concentration of 0.5-20.0 m L/L (P <0.05), the inhibition rate to NPYR reaches the highest value (L%) when the volume concentration of rosemary essential oil is 20.0m L/L, the inhibition rate reaches 84.16% when the volume concentration of rosemary essential oil is 20.0m L/L, the inhibition rate to Zanthoxylum bungeanum is in the range of 0.5-20.0 m L/L, the inhibition rate to Zanthoxylum piperitum is in the range of 0.5-L% and the volume concentration of Zanthoxylum piperitum is in the range of < 7.0.0.72%, the volume concentration of Zanthoxylum piperitum, the inhibition rate is increased from 0.72 to L% and the volume concentration of Zanthoxylum piperitum no < L, the inhibition rate is decreased from 0.72% to L% when the volume concentration of Zanthoxylum piperitum < 0.72, the volume concentration of Zanthoxylum piperitum < 0.72, the volume concentration, the NPY0/L, the NPY0% is increased from the NPM L, the NPK < 0% and the volume concentration of the NPK < 0% after the NPK < 0% of the NPK < 0/L, the NPK 0% of the NPK 0, the NPK 0/L, the NPK 0% of the NPK 0, the NPK 0% of the Zanthoxylum piper pepper is increased, the NPK 0/L, the NPK 0% of the NPK 0/L, the NPK 0/L, the NPK 0% of the NPK 0/L, the Zanthoxylum piper oil is increased, the NPK 0/L, the Zanthoxylum piper pepper is increased, the NPK 0/L, the NPK 72, the NP.
The SEO has a mellow flavor, a certain SEO only needs to be used at a high concentration (except the zanthoxylum oil, the aniseed oil, the rosemary oil and the clove oil are required to be more than 15.0m L/L in volume and shown in table 2) to inhibit NPYR to reach the maximum value in a group, however, in practical application, the single SEO with the high concentration cannot be added, the inhibiting rate of the angelica flavor ginger oil to NPYR is higher than that of the aniseed oil in each volume concentration, but the angelica is slightly bitter in flavor when being added into meat products, and the angelica is a spice commonly used in meat product processing and has wide application value, so that the final determination is that in the central combination test design of a response surface, a response surface optimization test is carried out by taking the zanthoxylum oil, the aniseed oil, the hemp oil, the clove oil and the rosemary oil as self variables, the concentration of the 5 SEO is determined at a zero level, mainly according to the principle that the SEO, clove and clove essential oils are selected as a final extraction factor of a final volume of 3975.2. the final combined essential oil, a volume of the SEO concentration is selected as a single zanthoxylum oil, a theoretical essential oil of 3913, a theoretical essential oil of a.
2.2 center of response surface combination test
Based on a single-factor test result, 5 factors of zanthoxylum essential oil (A), aniseed essential oil (B), pimento essential oil (C), clove essential oil (D) and rosemary essential oil (E) are selected as independent variables, and a Design-Expert7.0 software is used to carry out a 5-factor-3-level response surface optimization analysis test through a Box-Behnken central combined test Design principle, so as to research the influence of the Design on the NPYR inhibition rate (Y).
2.2.1 regression model construction and significance testing
The design and results of the response surface center combination test are shown in table 3.
The software Design-expert7.0 is used for carrying out multiple linear regression analysis on the test results, and a quadratic regression equation of the inhibition rate (Y) on the pepper essential oil (A), the aniseed essential oil (B), the pimento essential oil (C), the clove essential oil (D) and the rosemary essential oil (E) can be established:
Y＝70.01+0.013A+0.022B+0.026C+0.031D+0.030E+0.058AB+0.085AC+0.017AD+0.058AE-0.037BC+0.075BD+0.010BE+2.500E-003+0.035CE+0.043DE-0.55A2-0.54B2-0.51C2-0.54D2-0.55E2。
TABLE 3 center combination test design and results for response surface
Analysis of variance was performed on the regression model established in the experiment and the results are shown in table 4. As can be seen from table 4, model F is 78.98, P<0.01, reaching a very significant level, indicating that the regression model is meaningful; the mismatching term F is 1.51, and P is 0.3432>0.05, the model is not remarkably missimulated, which shows that the influence of unknown factors on the result is small, the test error mainly comes from random errors, the regression model established by the response surface method has high fitting degree, and the response surface can be really fitted. Determining coefficient R of model20.9433, the regression model established by the response surface method has better regression effect; adjustment factor Radj20.9720, which shows that the model can explain the change of 97.20% response value, so that the fitting degree and the reliability are high, the model can be used for analyzing and predicting the NPYR inhibition rate. CV (coefficient of variation of Y) represents the accuracy of the test, the lower the CV value, the higher the accuracy of the test, the lower the CV value of the test, which is 0.091%, the higher the accuracy of the explanation model, and the higher the reliability of the test operation, which has certain practical guiding significance, and can be used to theoretically predict the NPYR inhibition rate. In addition, the significance test of the regression equation coefficient on the model shows that the primary terms in the model are not significant, and the secondary terms are extremely significant (P)<0.01), AC and BD in interactive items reach a significant level (P)<0.05), the other items have no significant influence on the index (P)>0.05). Therefore, the influence of each factor on the NPYR inhibition ratio is not a simple linear relationship. According to the magnitude of the F value, the sequence of the NPYR inhibition effect of the five test factors is as follows: clove essential oil (D)>Rosemary essential oil (E)>Spicy pepper essential oil (C)>Octagonal essential oil (B)>And (C) pepper essential oil (A).
TABLE 4 response surface test regression model analysis of variance
Note: the P value is the result of the homogeneity check of the variance; indicates extreme significance at the a-0.01 level and indicates significance at the a-0.05 level.
2.2.2 response surface and contour plot analysis
The influence of research factors and interaction thereof on the NPYR inhibition rate can be reflected more visually through a response surface and a contour map drawn by Design-expert7.0 software, and the larger the curve curvature in the response surface map is, the larger the influence of the research factors on the NPYR inhibition rate is; the contour lines appear as ovals, indicating significant interaction between the factors studied, while circles indicate insignificant interaction. The response surface analysis of the interaction between the factors by fitting the regression equation of the model is shown in FIGS. 1-4. The interaction between the zanthoxylum essential oil and the zanthoxylum essential oil is obvious, after the volume concentration of the zanthoxylum essential oil is fixed, the NPYR inhibition rate shows a trend of increasing first and then decreasing along with the increase of the volume concentration of the zanthoxylum essential oil, the contour diagram is oval, and when the volume concentration of the zanthoxylum essential oil is fixed, the NPYR inhibition rate also increases first and then decreases along with the increase of the volume concentration of the zanthoxylum essential oil. The interaction between the anise essential oil and the clove essential oil also reaches a remarkable level (P is less than 0.05), when the anise essential oil is at a certain volume concentration, the NPYR inhibition effect is firstly increased and then reduced along with the increase of the volume concentration of the clove essential oil, and otherwise, the same change trend is presented. Therefore, the inhibition effect of the interaction between the zanthoxylum essential oil and the zanthoxylum essential oil, and the aniseed essential oil and the clove essential oil on NPYR is large, and the analysis result is consistent with the analysis result of variance.
2.2.3 optimum Process Condition prediction and verification test
The optimal proportion of CSEO for inhibiting NPYR is predicted according to Design-Expert7.0 software and is 441.0 mu L/L of pepper essential oil, 337.0 mu L/L of anise essential oil, 17.0 mu L/L of pepper essential oil, 14.0 mu L/L of clove essential oil and 137.0 mu L/L of rosemary essential oil, at the moment, the prediction result of the NPYR inhibition rate is 69.84%, the CSEO under the condition is verified in a system for simulating NPYR formation in vitro, the NPYR inhibition rate is (68.40 +/-0.33%) measured by the test and basically accords with the predicted value, the model parameters obtained by optimizing the application response surface method are accurate and reliable, the test has practical application value, the optimal proportion is verified in the system for simulating NDMA formation in vitro, the test is determined, the NDMA inhibition rate is (3 +/-0.43%), the influence of the nitrosation inhibitor at the same level is added into the system for simulating NDMA formation in the production, the west root is reasonably determined by the test, the NDMA inhibition rate is (66.69 +/-0.43%), the NDMA inhibition rate is reasonably determined by the test and the influence of the nitrosation inhibitor is determined by the test and the test on the nitrosation rate of the NPMA formation in the group, the test and the test, the influence of the nitrosation rate is determined by the test, the influence of the nitrosation rate of the nitrosation inhibitor in the nitrosation rate is determined by the test and the test, the nitrosation rate of the test is determined by the test, the test.
Although the test uses the optimum mixture ratio obtained by the combination design of the center of the response surface, the inhibition ratio of the NPYR is (68.40 ± 0.33)% (verification test value), although the result is slightly lower than the highest NPYR inhibition ratio in the single factor test (the volume concentration of rosemary is 20.0m L/L, and the inhibition ratio of the NPYR is 85.70%), because the volume concentration of each SEO in the screened optimum mixture ratio is far lower than the highest concentration when a single SEO is used in consideration of the influence of the addition amount of the SEO on the flavor of the meat product in the actual production and processing of the meat product.
3 conclusion
The results of single-factor tests show that in an in vitro simulated nitrosation system, except the amomum villosum essential oil, other 7 SEOs have different degrees of inhibition effects on the generation of NPYR, and the inhibition rate and the addition concentration thereof have a certain relationship, wherein the rosemary essential oil has the strongest inhibition effect on the NPYR, when the volume concentration of the rosemary essential oil is 20.0m L/L, the inhibition rate on the NPYR reaches the highest value (85.70%), the pepper essential oil (5.0-10.0 m L0/L1), the pepper essential oil (0.5m L/L) and the clove essential oil (15.0m L/L) have the inhibition rate on the NPYR of more than 68%, the angelica flavor ginger oil (10.0m L/L) and the aniseed essential oil (15.0m L/L) have the low inhibition effects (more than 50%), and the black pepper essential oil has the relatively weak inhibition rate on the NPYR (less than 50%).
The CSEO combined by 5 SEOs is obtained by a response surface center combination design test, the optimal volume concentration ratio is that when the volume concentrations of the zanthoxylum bungeanum essential oil, the aniseed essential oil, the pimento essential oil, the clove essential oil and the rosemary essential oil are 441.0, 337.0, 17.0, 14.0 and 137.0 mu L/L respectively, the predicted value of the NPYR inhibition rate is 69.84% preferably obtained by a model, the inhibition rate obtained by a verification test is (68.40 +/-0.33)%, the inhibition rate of the proportion on NDMA is (66.69 +/-0.43)%, the inhibitor of the level is added into a meat product (Western bacon), the NPYR inhibition rate and the NDMA inhibition rate are 66.13% and 65.99% respectively, the regression model obtained is good in fitting effect with the actual effect, has practical value, and provides a new theoretical basis and data support for the selection of the N-nitrosamine inhibitor in the meat product industry.
Although the embodiments of the present invention have been disclosed for illustrative purposes, those skilled in the art will appreciate that: various substitutions, changes and modifications are possible without departing from the spirit and scope of the invention and the appended claims, and therefore the scope of the invention is not limited to the embodiments disclosed.
Claims (10)
1. An in-vitro nitrosation simulation system establishment method for blocking NPYR generation by CSEO proportion in salted meat products is characterized in that: the specific method comprises the following steps:
⑴ in vitro simulation of nitrosation reaction
a, respectively sucking SEO with different volumes into a 50m L centrifugal tube, adding 0.05g PEG8000, and then adding 0.5 mol/L sodium citrate-hydrochloric acid buffer solution to make the total volume of the SEO to be 10.0m L;
b, homogenizing and emulsifying the solution obtained in the step a for 1min, putting 1.0m L prepared emulsion into a 50m L centrifuge tube, sequentially adding 0.5 mol/L sodium citrate-hydrochloric acid buffer solution 5.0m L, 1 mmol/L sodium nitrite 0.5m L, 1 mmol/L pyrrolidine hydrochloride 0.5m L and 3m L buffer solution to enable the total volume of the reaction system to be 10.0m L, and carrying out water bath reaction for 1h at 37 ℃;
⑵ NPYR determination
a, after the in vitro simulation nitrosation reaction in the step ⑴ is finished, taking 1.0m L reaction solution into a glass test tube, adding 0.5% sodium carbonate solution 0.5m L to terminate the reaction, and placing the glass test tube under a 15W ultraviolet lamp in a super clean bench for irradiating for 15 min;
b, taking out, immediately and sequentially adding 1% of sulfanilic acid 1.5m L, 0.1% of α -naphthylamine 1.5m L and 0.5m L buffer solution until the volume of the solution is accurate to 5.0m L, and shaking up;
c, standing for 15min for color development, and then carrying out colorimetric determination on the absorbance at the wavelength of 525 nm;
⑶ one-factor test
a, sucking 0.00, 0.05, 0.50, 1.00, 1.50 and 2.00m L for each SEO respectively to ensure that the final concentration of each SEO in the in-vitro simulated nitrosation reaction is 0.0, 0.5, 5.0, 10.0, 15.0 and 20.0m L/L respectively;
b, after the reaction is finished, carrying out NPYR determination, wherein a blank control group is blank emulsion without SEO;
c, calculating the inhibition rate of each SEO on NPYR at the corresponding volume concentration, and repeating the test for 3 times;
⑷ center of response surface combination test
On the basis of a single-factor test result, the SEO volume concentration is screened, the SEO volume concentration is taken as an influence factor level, the NPYR inhibition rate is taken as a response value, Design-Expert7.0 software is used, and a response surface optimization analysis test is designed according to a Box-Behnken center combined test Design principle;
⑷ validation test
Verification in vitro nitrosation simulation emulsification system
Adding the CSEO proportion obtained according to the design of the response surface center combined test into a system formed by in vitro simulation NPYR, and performing 6 times of repeated verification tests;
⑸ calculation of NPYR and NDMA inhibition rates
(A0Is the absorbance value of the blank emulsion without SEO; a. thexAs absorbance value upon addition of SEO emulsion).
⑹ data processing.
2. The method for establishing the in vitro nitrosation simulation system for blocking NPYR generation by CSEO ratio in the salted meat product as claimed in claim 1, wherein the pH of the sodium citrate-hydrochloric acid buffer solution in step ⑴ is 3.0.
3. The method for building the in-vitro nitrosation simulation system for the CSEO ratio blocking NPYR generation in the salted meat product as claimed in claim 1, wherein the homogenizing emulsification treatment in step ⑴ adopts a homogenizer, and the B shift rotation speed is 13000 r/min.
4. The method for building an in vitro nitrosation simulation system for generating CSEO proportion blocking NPYR in salted meat products as claimed in claim 1, wherein the distance between the liquid level of the test tube and the light tube in step ⑵ is 15 cm.
5. The method for establishing the in-vitro nitrosation simulation system for the CSEO ratio blocking NPYR generation in the salted meat product as claimed in claim 1, wherein in the single-factor test, the SEOs are 8 types, and the SEOs are 20.0m L/L volume concentration, 5.0-10.0 m L0/L1 volume concentration, 0.5m L/L volume concentration and 15.0m L/L volume concentration, 10.0m L/L volume concentration and 15.0m L/L volume concentration of the rosemary essential oil.
6. The method for establishing the in vitro nitrosation simulation system for the CSEO ratio blocking NPYR generation in the salted meat product as claimed in claim 5, is characterized in that the inhibition effect of the nitrosation inhibitor on NPYR and NDMA is analyzed by using the volume concentration of zanthoxylum essential oil (A, 330.0-470.0 mu L/L), the volume concentration of aniseed essential oil (B, 300.0-370.0 mu L/L), the volume concentration of pimento essential oil (C, 13.0-20.0 mu L/L), the volume concentration of clove essential oil (D, 10.0-17.0 mu L/L) and the volume concentration of rosemary essential oil (E, 100.0-170.0 mu L/L) as the influence factor level and using the NPYR inhibition rate as the response value, wherein the factor level is as follows:
7. the method for establishing the in vitro nitrosation simulation system for blocking NPYR generation by CSEO ratio in the salted meat product as claimed in claim 1, wherein: the CSEO proportion obtained by the central combination design of the response surface is applied to the Western-style bacon, the inhibition effect of the addition of the nitrosation inhibitor on NPYR and NDMA in the Western-style bacon is analyzed, meanwhile, a control experiment is carried out, and the test result of the combination design is further verified.
8. The method for establishing the in vitro nitrosation simulation system for blocking NPYR generation by CSEO proportion in the salted meat product as claimed in claim 7, is characterized in that the volume concentrations of the zanthoxylum bungeanum essential oil, the anise essential oil, the pimento essential oil, the clove essential oil and the rosemary essential oil are 441.0, 337.0, 17.0, 14.0 and 137.0 μ L/L respectively, the predicted value of the NPYR inhibition rate obtained by a model is 69.84%, the inhibition rate obtained by a verification test is (68.40 +/-0.33%), the inhibition rate of the proportion on NDMA is (66.69 +/-0.43%), and the inhibition rate of the NPYR and the NDMA obtained by adding the inhibitor at the level into the meat product (Western bacon) is 66.13% and 65.99% respectively.
9. The method for establishing the in vitro nitrosation simulation system for blocking NPYR generation by CSEO ratio in the salted meat product as claimed in claim 1, wherein: the data processing was performed by preprocessing the experimental data with Microsoft Excel 2010, data analysis with Statistix8.1, significant differences (P <0.05) by Turkey test program, sigmaplott 10.0 mapping.
10. The method for establishing the in vitro nitrosation simulation system for blocking NPYR generation by CSEO ratio in the salted meat product as claimed in claim 1, wherein: the response surface test is designed by adopting Design-Expert7.0 and results are analyzed, and the interaction effect of any two factors is analyzed and evaluated by establishing a regression equation, drawing contour lines and a response surface graph.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010294693.5A CN111398265A (en) | 2020-04-15 | 2020-04-15 | Establishment method of in-vitro nitrosation simulation system for blocking NPYR generation by CSEO proportion in salted meat product |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010294693.5A CN111398265A (en) | 2020-04-15 | 2020-04-15 | Establishment method of in-vitro nitrosation simulation system for blocking NPYR generation by CSEO proportion in salted meat product |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN111398265A true CN111398265A (en) | 2020-07-10 |
Family
ID=71433270
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202010294693.5A Pending CN111398265A (en) | 2020-04-15 | 2020-04-15 | Establishment method of in-vitro nitrosation simulation system for blocking NPYR generation by CSEO proportion in salted meat product |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN111398265A (en) |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106309521A (en) * | 2016-08-30 | 2017-01-11 | 甘肃中医药大学 | Method for optimizing process parameters of angelica sinensis total flavonoid extraction with response surface methodology |
| CN109679868A (en) * | 2018-12-28 | 2019-04-26 | 天津农学院 | A kind of microorganism nitrosation inhibitor and preparation method thereof |
| CN110057766A (en) * | 2019-05-06 | 2019-07-26 | 吉林工商学院 | Response surface optimization Semen Coicis polyphenol extracting method |
| CN110214921A (en) * | 2019-06-20 | 2019-09-10 | 天津农学院 | A kind of aromatic condiment essential oil composite inhibitor for blocking NDMA to generate in in-vitro simulated nitrosation system |
-
2020
- 2020-04-15 CN CN202010294693.5A patent/CN111398265A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106309521A (en) * | 2016-08-30 | 2017-01-11 | 甘肃中医药大学 | Method for optimizing process parameters of angelica sinensis total flavonoid extraction with response surface methodology |
| CN109679868A (en) * | 2018-12-28 | 2019-04-26 | 天津农学院 | A kind of microorganism nitrosation inhibitor and preparation method thereof |
| CN110057766A (en) * | 2019-05-06 | 2019-07-26 | 吉林工商学院 | Response surface optimization Semen Coicis polyphenol extracting method |
| CN110214921A (en) * | 2019-06-20 | 2019-09-10 | 天津农学院 | A kind of aromatic condiment essential oil composite inhibitor for blocking NDMA to generate in in-vitro simulated nitrosation system |
Non-Patent Citations (3)
| Title |
|---|
| 熊凤娇 等: "二次回归正交设计优选阻断NDMA形成的亚硝化抑制剂", 《肉类研究》 * |
| 陈文静 等: "体外模拟亚硝化体系中优选阻断NDMA生成的香辛料精油复合抑制剂", 《食品工业科技》 * |
| 陈文静 等: "复合香辛料亚硝化抑制剂对西式培根品质的影响", 《肉类研究》 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Uebelacker et al. | Quantitative determination of acetaldehyde in foods using automated digestion with simulated gastric fluid followed by headspace gas chromatography | |
| Millour et al. | Simultaneous analysis of 21 elements in foodstuffs by ICP-MS after closed-vessel microwave digestion: Method validation | |
| Villamor et al. | Effects of ethanol, tannin, and fructose concentrations on sensory properties of model red wines | |
| Fernández de Simón et al. | Phenolic compounds in a Spanish red wine aged in barrels made of Spanish, French and American oak wood | |
| CN108918796A (en) | A method of cherry wine Ester fragrance synergistic action effect is analyzed based on σ-τ intensity method | |
| JP2013128464A (en) | Fermented malt beverage imparted with fruity fragrance | |
| CN108761002A (en) | A method of alcohols in chrysanthemum essential oil is analyzed based on S curve method and is acted synergistically with terpenoid substance fragrance | |
| Ross et al. | Odour detection threshold and consumer rejection of 1, 1, 6‐trimethyl‐1, 2‐dihydronaphthalene in 1‐year‐old R iesling wines | |
| CN108982755A (en) | A method of based on S curve method analysis cherry wine Ester fragrance synergistic effect | |
| CN104267164B (en) | A kind of method of easy Fast Measurement yellow rice wine alcoholic strength | |
| Mönch et al. | Folate bioavailability from foods rich in folates assessed in a short term human study using stable isotope dilution assays | |
| CN103217413A (en) | Analysis method for determining potassium in potassium metavanadate and/or sodium in sodium metavanadate | |
| Darias-Martı́n et al. | Comparative study of methods for determination of titrable acidity in wine | |
| Cooper et al. | Impact of light source on color and lipid oxidative stabilities from a moderately color-stable beef muscle during retail display | |
| CN103487446B (en) | A kind of based on the detection method of Alumen additive in the fried food of dielectric property | |
| CN111398265A (en) | Establishment method of in-vitro nitrosation simulation system for blocking NPYR generation by CSEO proportion in salted meat product | |
| Qian et al. | Decoding the aroma characteristics of icewine by partial least-squares regression, aroma reconstitution, and omission studies | |
| Honorato et al. | Analytical curve or standard addition method: how to elect and design—a strategy applied to copper determination in sugarcane spirits using AAS | |
| AUBRY et al. | Sensory analysis of Burgundy Pinot noir wines: A comparison of orthonasal and retronasal profiling | |
| CN110214921A (en) | A kind of aromatic condiment essential oil composite inhibitor for blocking NDMA to generate in in-vitro simulated nitrosation system | |
| CN109060747B (en) | Method for rapidly detecting sulfur dioxide in white granulated sugar | |
| CN105424790B (en) | A kind of method of external source addition ethyl lactate in detection white wine | |
| CN105424897B (en) | A kind of method of external source addition ethyl hexanoate in detection white wine | |
| Jack | Development of guidelines for the preparation and handling of sensory samples in the Scotch whisky industry | |
| Cliff et al. | Comparison of new and existing threshold methods for evaluating sulfur compounds in different base wines |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20200710 |