CN105717531A - Method for detecting electron beam irradiation dose in aquatic products - Google Patents
Method for detecting electron beam irradiation dose in aquatic products Download PDFInfo
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- CN105717531A CN105717531A CN201610088219.0A CN201610088219A CN105717531A CN 105717531 A CN105717531 A CN 105717531A CN 201610088219 A CN201610088219 A CN 201610088219A CN 105717531 A CN105717531 A CN 105717531A
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Abstract
The invention relates to a method for detecting an electron beam irradiation dose in aquatic products. The method comprises the steps of: establishing a linear equation between O-tyrosine content in the electron beam irradiation aquatic products and the electron beam irradiation dose; utilizing a high performance liquid chromatography to detect the O-tyrosine content in the electron beam irradiation aquatic products; and obtaining the electron beam irradiation dose through the linear equation. According to the invention, the irradiation dose is determined by detecting the amount of the O-tyrosine content after the electron beam irradiation, and a theoretical basis for the safety of irradiated food is provided. The method for detecting the electron beam irradiation dose in the aquatic products has the advantages that the detecting method is rapid, convenient, loseless and high in precision; the detection method has a good linear relation with a target compound, the recovery rate is high, the relative deviation is small, the used reagent is less toxic, the usage amount is little, and the detecting method is environmentally friendly.
Description
Technical field
The present invention relates to a kind of analysis method of aquatic product quality and safety, particularly relate to a kind of method of electron beam irradiation dosage in easy and simple to handle, detection aquatic products rapidly and efficiently.
Background technology
Food irradiation technique is increasingly widely applied in food industry as a class cold processing technique, irradiated food carries out effectively supervision and is not only the eager desire of consumer, and be ensure the inevitable requirement that irradiated food develops in a healthy way.The detection method of current irradiated food is mainly to be set up based on gamma-ray irradiation, and on the products such as the spice that focuses mostly on, meat of poultris, dry fruit.
The effect of food is included primary effect and secondary action by radiation sterilizing, and primary effect is ionization and the chemical action that microbial cell matter interstitial is subject to generation after high-energy irradiation, makes material formation ion, excited state and molecular fragment；Secondary action be moisture etc. irradiated there is ionization and produce H, OH free radical and peroxide again with other material effects in cell, there is cross-linking reaction.Both effects cause the change of chemical bond in the organic molecules such as microbial DNA, RNA, protein, lipid, protein and DNA molecular crosslinking, the change of base in DNA sequence, thus causing the death of microbial cell, reaches to extend food storage time and fresh-keeping purpose.
There is dispute in the potential safety issue of irradiated food, the FAO/IEAE/WHO food irradiation joint specialist committee held in Geneva points out that " population mean dosage is any food of below 10kGy irradiation, it does not have toxicological critical, it is not necessary to carry out toxicological evaluation.Simultaneously on threpsology and microbiology is also safe ".But the color of food, taste, nutrition can be produced some side effect by irradiation improperly, the change of such as frowziness, color and luster and the destruction of nutritional labeling, the health of people be there is also potential safety hazard by the existence of radiolysis products.
Detection method for irradiated food generally has ESR electron spin resonance, thermoluminescence, DNA Comet Assay and adjacent tyrosine detection method etc., but ESR electron spin resonance detection equipment cost is higher, testing result needs professional to resolve, and limits its extensive use to a certain extent；Thermo luminescence method complex operation, sample size and type requirements is higher, biased sample sensitivity is relatively low；The accuracy of DNA Comet Assay detection needs to be investigated.
Both at home and abroad it is changed to starting point with compositions such as lipid, protein, carbohydrate, DNA after predose, carry out the exploitation of a series of detection technique, wherein adjacent tyrosine is a kind of marker being transformed by phenylalanine in radiative process, may be used for the detection of the higher irradiated food of protein content, but current research major part is for the detection of gamma-ray irradiation aquatic products, and the relation of adjacent tyrosine with irradiation dose is still not clear.
Summary of the invention
It is an object of the invention to there is no a defect of method fast and effectively and a kind of method of electron beam irradiation dosage in easy and simple to handle, detection aquatic products rapidly and efficiently is provided to solve existing detection electron beam irradiation dosage.
To achieve these goals, the present invention is by the following technical solutions:
A kind of detect the method for electron beam irradiation dosage in aquatic products, described method is first build in electron beam irradiation aquatic products the linear equation between adjacent tyrosine content and electron beam irradiation dosage, then utilize after high performance liquid chromatography detection electron beam irradiation the content of adjacent tyrosine in aquatic products, obtain electron beam irradiation dosage finally by linear equation.In the technical program, in food, phenylalanine is at the hydroxyl radical reaction of the rainy irradiation generation of effect of irradiation, produce tyrosine and two kinds of tyrosine isomers, i.e. adjacent tyrosine (Ortho-tyrosine, it is abbreviated as o-tyrosine) and m-Tyrosine (meta-tyrosine, it is abbreviated as m-tyrosine), tyrosine exists and body itself, therefore adjacent tyrosine and m-Tyrosine are often used as irradiation marker to differentiate that whether food is through irradiation, and adjacent tyrosine and m-Tyrosine are due to the similar more difficult separation of polarity, therefore neighbour's tyrosine is commonly used for the detection of irradiated food.The present invention by detect adjacent tyrosine content after electron beam irradiation number determine irradiation dose, the safety for irradiated food is provided fundamental basis, and the detection method of the present invention is fast and convenient, degree of accuracy is high.
As preferably, comprising the following steps:
A) the choosing and pretreatment of raw material: take scallop, base encloses the electron beam irradiation that shrimp and Lateolabrax japonicus (Cuvier et Va-lenciennes) (Lateolabracis) carry out 1-10kGy irradiation dose and processes, homogenate is placed at-40 DEG C preservation 5-10h；
B) pre-treatment: take step a) the sample 1-1.2g obtained, it is placed in tool plug centrifuge tube, adding 5-6.5mL mass fraction is 0.2% aqueous formic acid, 20000-22000r/min homogenizing 4-5min, ultrasonic 20-35min, centrifugal 25-30min at 10000-12000rpm4 DEG C, take supernatant, add 10-15mL acetone, place 3-5h for-40 DEG C, centrifugal 8-12min at 10000-12000rpm4 DEG C, take supernatant in 50-55 DEG C of outstanding steaming, lyophilizing after removing organic solvent, be that 0.2% formic acid redissolves with 1-1.2mL mass fraction, cross 0.22 μm of organic filter membrane, upper machine testing.
As preferably, the chromatographic condition of high performance liquid chromatography is: chromatographic column: AglientEclipse-C18 post (4.6mm × 250mm, 5.0 μm) chromatographic column；Mobile phase: A:0.1% formic acid, B: ethanol (20:80, v/v, in ethanol containing volume ratio be 0.5% glacial acetic acid and 0.3% triethylamine)；Column temperature: 30 DEG C, wavelength: Ex:275nm, Em:305nm, sample size: 20 μ L.
As preferably, Lateolabrax japonicus (Cuvier et Va-lenciennes) (Lateolabracis) is Y=0.1133X3-0.3534X2+ 5.2943X-1.1651, R2=0.9986；Wherein, Y is adjacent tyrosine content, and X is electron beam irradiation dosage.
As preferably, it is Y=-0.5626X that base encloses shrimp3+10.206X2-4.9731X+6.8588, R2=0.9938；Wherein, Y is adjacent tyrosine content, and X is electron beam irradiation dosage.
As preferably, scallop is Y=0.1376X respectively3-2.7062X2+ 19.949X+2.3847, R2=0.985；Wherein, Y is adjacent tyrosine content, and X is electron beam irradiation dosage.
As preferably, electron beam irradiation dosage respectively 1kGy, 3kGy, 5kGy, 7kGy, 10kGy in step a).
The invention has the beneficial effects as follows the present invention by detect adjacent tyrosine content after electron beam irradiation number determine irradiation dose, the safety for irradiated food is provided fundamental basis, and the detection method of the present invention is fast and convenient, lossless, degree of accuracy is high；Detection method is good to the linear relationship of target compound, and the response rate is high, and relative deviation is little, the reagent low toxicity of use, makes consumption few, environmentally friendly；Irradiation dose is in the scope of 1-10kGy, and the relation of the adjacent tyrosine content in three kinds of electron beam irradiation aquatic products and irradiation dose all can adopt cubic function to be simulated, its coefficient R2> 0.98.
Accompanying drawing explanation
Fig. 1 is the Linear equations of adjacent tyrosine content and irradiation dose in Lateolabrax japonicus (Cuvier et Va-lenciennes) (Lateolabracis) of the present invention.
Fig. 2 is that base of the present invention encloses the Linear equations of adjacent tyrosine content and irradiation dose in shrimp.
Fig. 3 is the Linear equations of adjacent tyrosine content and irradiation dose in scallop of the present invention.
Detailed description of the invention
Below by detailed description of the invention, the present invention will be further described.
In the present invention, if not refering in particular to, all devices and raw material all can be buied from market or the industry is conventional, and the method in following embodiment if no special instructions, is this area conventional method.
Scallop, base enclose shrimp, Lateolabrax japonicus (Cuvier et Va-lenciennes) (Lateolabracis) is purchased from the market of farm produce, South Mountain, Qingdao；
Agilent1260 high performance liquid chromatograph (is furnished with fluorescence detector) purchased from American Aglient company；
Vacuum rotary evaporator (Hei-Vap) is purchased from Heidolph company of Germany；
High-speed tissue mashing machine (DS-1) is purchased from Shanghai Sample Model Factory；
High speed dispersor is purchased from IKA instrument and equipment company limited；
High speed refrigerated centrifuge (3K-15) is purchased from Sigma company of Germany；
Ultrasonic cleaner (KQ5200B) is purchased from Kunshan Ultrasonic Instruments Co., Ltd.；
Electronic analytical balance (precision 0.1g, TE601-2) is purchased from Sai Duolisi company limited of Germany；
Electronic analytical balance (precision 0.0001g, BS-224-S) is purchased from Sai Duolisi company limited of Germany；
Ultrapure water system (Milli-Q) purchased from American Millipore company；
Magnetic stirring apparatus (GL-3250A) is purchased from Beijing LabTech Instrument Ltd.；
AglientEclipse-C18 post (4.6mm × 250mm, 5.0 μm) purchased from American Aglient company；Glacial acetic acid (analytical pure), triethylamine (analytical pure) are purchased from Chemical Reagent Co., Ltd., Sinopharm Group；
DL-neighbour's tyrosine (DL-o-tyrosine) available from Sigma, purity >=96.0%.
The chromatographic condition of high performance liquid chromatography is: chromatographic column: AglientEclipse-C18 post (4.6mm × 250mm, 5.0 μm) chromatographic column；Mobile phase: A:0.1% formic acid, B: ethanol (20:80, v/v, in ethanol containing volume ratio be 0.5% glacial acetic acid and 0.3% triethylamine)；Column temperature: 30 DEG C, wavelength: Ex:275nm, Em:305nm, sample size: 20 μ L.
Gradient Elution program: 0-14min, 92% (A)；14-14.2min, 92% (A)-20% (A)；14.2-21min, 20% (A)；21-21.2min, 20% (A)-92% (A)；21.2-30min, 92% (A).
Embodiment 1
A kind of detect the method for electron beam irradiation dosage in aquatic products, comprise the following steps:
A) the choosing and pretreatment of raw material: take scallop, base encloses the electron beam irradiation that shrimp and Lateolabrax japonicus (Cuvier et Va-lenciennes) (Lateolabracis) carry out 1kGy, 3kGy, 5kGy, 7kGy, 10kGy irradiation dose respectively and processes, and homogenate preserves 5h at being placed on-40 DEG C；
B) pre-treatment: take step a) the sample 1g obtained, is placed in tool plug centrifuge tube, and adding 5mL mass fraction is 0.2% aqueous formic acid, 20000r/min homogenizing 4min, centrifugal 25min at ultrasonic 20min, 10000rpm4 DEG C, take supernatant, add 10mL acetone, place 3h for-40 DEG C, centrifugal 8-12min at 10000rpm4 DEG C, take supernatant in 50 DEG C of outstanding steamings, lyophilizing after removing organic solvent, be that 0.2% formic acid redissolves with 1mL mass fraction, cross 0.22 μm of organic filter membrane, upper machine testing.
Embodiment 2
A kind of detect the method for electron beam irradiation dosage in aquatic products, comprise the following steps:
A) the choosing and pretreatment of raw material: take scallop, base encloses the electron beam irradiation that shrimp and Lateolabrax japonicus (Cuvier et Va-lenciennes) (Lateolabracis) carry out 1kGy, 3kGy, 5kGy, 7kGy, 10kGy irradiation dose respectively and processes, and homogenate preserves 8h at being placed on-40 DEG C；
B) pre-treatment: take step a) the sample 1.1g obtained, is placed in tool plug centrifuge tube, and adding 5.5mL mass fraction is 0.2% aqueous formic acid, 21000r/min homogenizing 4.5min, centrifugal 28min at ultrasonic 25min, 11000rpm4 DEG C, take supernatant, add 12mL acetone, place 4h for-40 DEG C, centrifugal 10min at 11000rpm4 DEG C, take supernatant in 53 DEG C of outstanding steamings, lyophilizing after removing organic solvent, be that 0.2% formic acid redissolves with 1.1mL mass fraction, cross 0.22 μm of organic filter membrane, upper machine testing.
Embodiment 3
A kind of detect the method for electron beam irradiation dosage in aquatic products, comprise the following steps:
A) the choosing and pretreatment of raw material: take scallop, base encloses the electron beam irradiation that shrimp and Lateolabrax japonicus (Cuvier et Va-lenciennes) (Lateolabracis) carry out 1kGy, 3kGy, 5kGy, 7kGy, 10kGy irradiation dose respectively and processes, and homogenate preserves 10h at being placed on-40 DEG C；
B) pre-treatment: take step a) the sample 1.2g obtained, is placed in tool plug centrifuge tube, and adding 6.5mL mass fraction is 0.2% aqueous formic acid, 22000r/min homogenizing 5min, centrifugal 30min at ultrasonic 35min, 12000rpm4 DEG C, take supernatant, add 15mL acetone, place 5h for-40 DEG C, centrifugal 12min at 12000rpm4 DEG C, take supernatant in 55 DEG C of outstanding steamings, lyophilizing after removing organic solvent, be that 0.2% formic acid redissolves with 1.2mL mass fraction, cross 0.22 μm of organic filter membrane, upper machine testing.
Comparative example 1, takes scallop, base encloses shrimp and Lateolabrax japonicus (Cuvier et Va-lenciennes) (Lateolabracis) without electron beam irradiation, and all the other steps are all identical with embodiment 1.
Comparative example 1 does not detect adjacent tyrosine content.
Table 1 is the adjacent tyrosine content in Lateolabrax japonicus (Cuvier et Va-lenciennes) (Lateolabracis) under various dose in embodiment 1-3.
Adjacent tyrosine content in Lateolabrax japonicus (Cuvier et Va-lenciennes) (Lateolabracis) under table 1, various dose
In Lateolabrax japonicus (Cuvier et Va-lenciennes) (Lateolabracis), adjacent tyrosine content is shown in Fig. 1 with irradiation dose graph of a relation.
Lateolabrax japonicus (Cuvier et Va-lenciennes) (Lateolabracis) is Y=0.1133X3-0.3534X2+ 5.2943X-1.1651, R2=0.9986；Wherein, Y is adjacent tyrosine content, and X is electron beam irradiation dosage.
Table 2 is the adjacent tyrosine content that in embodiment 1-3, under various dose, base encloses in shrimp.
Under table 2, different irradiation dose, base encloses the adjacent tyrosine content in shrimp
The adjacent tyrosine content that base encloses in shrimp is shown in Fig. 2 with irradiation dose graph of a relation.
It is Y=-0.5626X that base encloses shrimp3+10.206X2-4.9731X+6.8588, R2=0.9938；Wherein, Y is adjacent tyrosine content, and X is electron beam irradiation dosage.
Table 3 is the adjacent tyrosine content in scallop under various dose in embodiment 1-3.
Adjacent tyrosine content in scallop under table 3, different irradiation dose
Adjacent tyrosine content in scallop and irradiation dose graph of a relation are shown in Fig. 3.
Scallop is Y=0.1376X respectively3-2.7062X2+ 19.949X+2.3847, R2=0.985；Wherein, Y is adjacent tyrosine content, and X is electron beam irradiation dosage.
Adjacent tyrosine content from the known three kinds of electron beam irradiation aquatic products of table 1-3 increases along with the rising of irradiation dose, and the adjacent tyrosine content in variety classes aquatic products is different, relevant to the structure of aquatic products self and character；Irradiation dose is in the scope of 1-10kGy, and the relation of the adjacent tyrosine content in three kinds of electron beam irradiation aquatic products and irradiation dose all can adopt cubic function to be simulated, and sees Fig. 1-3, its coefficient R2> 0.98.
Claims (7)
1. one kind is detected the method for electron beam irradiation dosage in aquatic products, it is characterized in that, described method is first build in electron beam irradiation aquatic products the linear equation between adjacent tyrosine content and electron beam irradiation dosage, then utilize after high performance liquid chromatography detection electron beam irradiation the content of adjacent tyrosine in aquatic products, obtain electron beam irradiation dosage finally by linear equation.
2. according to claim 1 a kind of detect the method for electron beam irradiation dosage in aquatic products, it is characterised in that comprise the following steps:
A) the choosing and pretreatment of raw material: take scallop, base encloses the electron beam irradiation that shrimp and Lateolabrax japonicus (Cuvier et Va-lenciennes) (Lateolabracis) carry out 1-10kGy irradiation dose and processes, homogenate is placed at-40 DEG C preservation 5-10h；
B) pre-treatment: take the sample 1-1.2g that step a) obtains, it is placed in tool plug centrifuge tube, adding 5-6.5mL mass fraction is 0.2% aqueous formic acid, 20000-22000r/min homogenizing 4-5min, ultrasonic 20-35min, centrifugal 25-30min at 10000-12000rpm4 DEG C, take supernatant, add 10-15mL acetone, place 3-5h for-40 DEG C, centrifugal 8-12min at 10000-12000rpm4 DEG C, take supernatant in 50-55 DEG C of outstanding steaming, lyophilizing after removing organic solvent, be that 0.2% formic acid redissolves with 1-1.2mL mass fraction, cross 0.22 μm of organic filter membrane, upper machine testing.
3. according to claim 1 and 2 a kind of detect the method for electron beam irradiation dosage in aquatic products, it is characterised in that the chromatographic condition of high performance liquid chromatography is: chromatographic column: AglientEclipse-C18 post (4.6mm × 250mm, 5.0 μm) chromatographic column；Mobile phase: A:0.1% formic acid, B: ethanol (20:80, v/v, in ethanol containing volume ratio be 0.5% glacial acetic acid and 0.3% triethylamine)；Column temperature: 30 DEG C, wavelength: Ex:275nm, Em:305nm, sample size: 20 μ L.
4. according to claim 1 and 2 a kind of detect the method for electron beam irradiation dosage in aquatic products, it is characterised in that Lateolabrax japonicus (Cuvier et Va-lenciennes) (Lateolabracis) is Y=0.1133X3-0.3534X2+ 5.2943X-1.1651, R2=0.9986；Wherein, Y is adjacent tyrosine content, and X is electron beam irradiation dosage.
5. according to claim 1 and 2 a kind of detect the method for electron beam irradiation dosage in aquatic products, it is characterised in that it is Y=-0.5626X that base encloses shrimp3+10.206X2-4.9731X+6.8588, R2=0.9938；Wherein, Y is adjacent tyrosine content, and X is electron beam irradiation dosage.
6. according to claim 1 and 2 a kind of detect the method for electron beam irradiation dosage in aquatic products, it is characterised in that scallop is Y=0.1376X respectively3-2.7062X2+ 19.949X+2.3847, R2=0.985；Wherein, Y is adjacent tyrosine content, and X is electron beam irradiation dosage.
7. according to claim 2 a kind of detect the method for electron beam irradiation dosage in aquatic products, it is characterised in that electron beam irradiation dosage respectively 1kGy, 3kGy, 5kGy, 7kGy, 10kGy in step a).
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Cited By (3)
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| CN106568870A (en) * | 2016-12-28 | 2017-04-19 | 舟山出入境检验检疫局综合技术服务中心 | Detection method of irradiated dried seafood |
| CN106597515A (en) * | 2016-11-28 | 2017-04-26 | 江苏省农业科学院 | Product irradiation dose determination method |
| CN108896679A (en) * | 2018-07-18 | 2018-11-27 | 精晶药业股份有限公司 | A kind of efficient liquid phase detection method of l-tyrosine |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN106597515A (en) * | 2016-11-28 | 2017-04-26 | 江苏省农业科学院 | Product irradiation dose determination method |
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| CN108896679A (en) * | 2018-07-18 | 2018-11-27 | 精晶药业股份有限公司 | A kind of efficient liquid phase detection method of l-tyrosine |
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