CN105717531A - Method for detecting electron beam irradiation dose in aquatic products - Google Patents
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- 238000010894 electron beam technology Methods 0.000 title claims abstract description 51
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- 235000019253 formic acid Nutrition 0.000 claims description 13
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 12
- ZMANZCXQSJIPKH-UHFFFAOYSA-N Triethylamine Chemical compound CCN(CC)CC ZMANZCXQSJIPKH-UHFFFAOYSA-N 0.000 claims description 12
- 230000005855 radiation Effects 0.000 claims description 12
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 claims description 10
- 239000006228 supernatant Substances 0.000 claims description 9
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims description 8
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- 239000002994 raw material Substances 0.000 claims description 6
- 239000007864 aqueous solution Substances 0.000 claims description 5
- 239000012528 membrane Substances 0.000 claims description 5
- 239000003960 organic solvent Substances 0.000 claims description 5
- 229960000583 acetic acid Drugs 0.000 claims description 4
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- 238000001514 detection method Methods 0.000 description 15
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- 239000000126 substance Substances 0.000 description 5
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- JZKXXXDKRQWDET-UHFFFAOYSA-N meta-tyrosine Natural products OC(=O)C(N)CC1=CC=CC(O)=C1 JZKXXXDKRQWDET-UHFFFAOYSA-N 0.000 description 4
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- OUYCCCASQSFEME-QMMMGPOBSA-N L-tyrosine Chemical compound OC(=O)[C@@H](N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-QMMMGPOBSA-N 0.000 description 2
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- 230000005251 gamma ray Effects 0.000 description 2
- 150000002632 lipids Chemical class 0.000 description 2
- COLNVLDHVKWLRT-UHFFFAOYSA-N phenylalanine Natural products OC(=O)C(N)CC1=CC=CC=C1 COLNVLDHVKWLRT-UHFFFAOYSA-N 0.000 description 2
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01T—MEASUREMENT OF NUCLEAR OR X-RADIATION
- G01T1/00—Measuring X-radiation, gamma radiation, corpuscular radiation, or cosmic radiation
- G01T1/02—Dosimeters
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Abstract
本发明涉及一种检测水产品中电子束辐照剂量的方法，所述方法为首先构建电子束辐照水产品中邻酪氨酸含量与电子束辐照剂量之间的线性方程，然后利用高效液相色谱法检测电子束辐照后水产品中邻酪氨酸的含量，最后通过线性方程得到电子束辐照剂量。本发明通过检测电子束辐照后邻酪氨酸含量的多少来确定辐照剂量，为辐照食品的安全性提供理论基础，本发明的检测方法快速简便、无损、精确度高；检测方法对目标化合物的线性关系好，回收率高，相对偏差小，使用的试剂低毒，使用量少，对环境友好。
The invention relates to a method for detecting electron beam irradiation dose in aquatic products. The method is to firstly construct a linear equation between the o-tyrosine content in electron beam irradiated aquatic products and the electron beam irradiation dose, and then use the high-efficiency The content of o-tyrosine in aquatic products after electron beam irradiation was detected by liquid chromatography, and finally the electron beam irradiation dose was obtained by linear equation. The invention determines the irradiation dose by detecting the content of o-tyrosine after electron beam irradiation, and provides a theoretical basis for the safety of irradiated food. The linear relationship of the target compound is good, the recovery rate is high, the relative deviation is small, the reagents used are low-toxic, the usage amount is small, and the environment is friendly.
Description
技术领域technical field
本发明涉及一种水产品质量与安全的分析方法，尤其涉及一种操作简便、快速高效的检测水产品中电子束辐照剂量的方法。The invention relates to an analysis method for the quality and safety of aquatic products, in particular to a method for detecting electron beam radiation dose in aquatic products with simple operation, fast and high efficiency.
背景技术Background technique
食品辐照技术作为一类冷加工技术越来越广泛的应用到食品工业中，对辐照食品进行有效监管不仅是消费者的迫切愿望，而且是保障辐照食品健康发展的必然要求。当前辐照食品的检测方法主要是γ射线辐照为基础建立起来的，而且多集中香辛料、禽肉类、干果类等产品上。As a kind of cold processing technology, food irradiation technology is more and more widely used in the food industry. Effective supervision of irradiated food is not only the urgent desire of consumers, but also an inevitable requirement to ensure the healthy development of irradiated food. The current detection methods for irradiated food are mainly established on the basis of γ-ray irradiation, and most of them are concentrated on spices, poultry meat, dried fruits and other products.
辐照杀菌对食品的作用包括初级作用和次级作用，初级作用即微生物细胞质间质受到高能照射后发生的电离作用和化学作用，使物质形成离子、激发态和分子碎片；次级作用是水分等经辐照发生电离而产生H、OH自由基和过氧化物再与细胞内其他物质作用，发生交联反应。这两种作用引起微生物DNA、RNA、蛋白质、脂类等有机分子中化学键的变化，蛋白质与DNA分子交联，DNA序列中碱基的改变，从而导致微生物细胞的死亡，达到延长食品储藏时间和保鲜的目的。The effect of radiation sterilization on food includes primary effect and secondary effect. The primary effect is the ionization and chemical action of the microbial cytoplasmic interstitial substance after being irradiated by high energy, which makes the substance form ions, excited states and molecular fragments; the secondary effect is moisture After ionization by irradiation, H, OH free radicals and peroxides are generated, and then interact with other substances in the cell to undergo a cross-linking reaction. These two effects cause changes in chemical bonds in organic molecules such as microbial DNA, RNA, proteins, and lipids, cross-linking of proteins and DNA molecules, and changes in bases in DNA sequences, which lead to the death of microbial cells and prolong food storage. The purpose of keeping fresh.
辐照食品的潜在安全性问题存在争议，在日内瓦召开的FAO/IEAE/WHO食品辐照联合专家委员会指出“总体平均剂量为10kGy以下辐照的任何食品，没有毒理学危险，无需进行毒理学评价。同时在营养学上和微生物学上也是安全的”。但不当的辐照会对食品的颜色、味道、营养产生一些副作用，比如有臭味、色泽的改变和营养成分的破坏，辐解产物的存在对人的健康也存在安全隐患。The potential safety of irradiated food is controversial. The FAO/IEAE/WHO Food Irradiation Joint Expert Committee held in Geneva pointed out that "any food irradiated with an overall average dose of less than 10kGy has no toxicological hazard and no toxicological evaluation is required. .It is also nutritionally and microbiologically safe". However, improper irradiation will cause some side effects on the color, taste, and nutrition of food, such as odor, color change, and destruction of nutritional components. The existence of radiolysis products also poses safety hazards to human health.
对于辐照食品的检测方法一般有ESR电子自旋共振、热释光、DNA彗星分析法与邻酪氨酸检测法等，但是ESR电子自旋共振检测设备成本较高，检测结果需要专业人员解析，一定程度上限制了它的广泛应用；热释光方法操作繁琐，对样品数量和种类要求较高，对混合样品敏感性较低；DNA彗星分析检测的准确性有待考究。The detection methods for irradiated food generally include ESR electron spin resonance, thermoluminescence, DNA comet analysis and o-tyrosine detection, etc. However, the cost of ESR electron spin resonance detection equipment is relatively high, and the detection results require professional analysis , which limits its wide application to a certain extent; the thermoluminescence method is cumbersome to operate, has high requirements on the number and type of samples, and is less sensitive to mixed samples; the accuracy of DNA comet analysis and detection needs to be studied.
国内外以辐照前后脂类、蛋白质、碳水化合物、DNA等成分的变化为出发点，进行了一系列检测技术的开发，其中邻酪氨酸是在辐射过程中由苯丙氨酸转化而成的一种标示物，可以用于蛋白质含量较高的辐照食品的检测，但是当前研究大部分针对γ射线辐照水产品的检测，而且对邻酪氨酸与辐照剂量的关系尚不明确。Based on the changes of lipids, proteins, carbohydrates, DNA and other components before and after irradiation, a series of detection technologies have been developed at home and abroad. Among them, o-tyrosine is converted from phenylalanine during the irradiation process. A marker that can be used for the detection of irradiated food with high protein content, but most of the current research is on the detection of γ-ray irradiated aquatic products, and the relationship between o-tyrosine and irradiation dose is not yet clear.
发明内容Contents of the invention
本发明的目的在于为了解决现有检测电子束辐照剂量没有快速有效的方法的缺陷而提供一种操作简便、快速高效的检测水产品中电子束辐照剂量的方法。The object of the present invention is to provide a simple, fast and efficient method for detecting electron beam radiation dose in aquatic products in order to solve the defect that there is no fast and effective method for detecting electron beam radiation dose in the prior art.
为了实现上述目的，本发明采用以下技术方案：In order to achieve the above object, the present invention adopts the following technical solutions:
一种检测水产品中电子束辐照剂量的方法，所述方法为首先构建电子束辐照水产品中邻酪氨酸含量与电子束辐照剂量之间的线性方程，然后利用高效液相色谱法检测电子束辐照后水产品中邻酪氨酸的含量，最后通过线性方程得到电子束辐照剂量。在本技术方案中，食品中苯丙氨酸在辐照的作用下雨辐照产生的羟基自由基反应，生产酪氨酸和两种酪氨酸异构体，即邻酪氨酸(Ortho-tyrosine，简写为o-tyrosine)和间酪氨酸(meta-tyrosine，简写为m-tyrosine)，酪氨酸存在与机体本身，因此邻酪氨酸和间酪氨酸常被用作辐照标识物来鉴别食品是否经过辐照，而邻酪氨酸和间酪氨酸由于极性相似较难分离，故邻酪氨酸常被用于辐照食品的检测。本发明通过检测电子束辐照后邻酪氨酸含量的多少来确定辐照剂量，为辐照食品的安全性提供理论基础，本发明的检测方法快速简便、精确度高。A method for detecting the dose of electron beam irradiation in aquatic products, the method is firstly constructing a linear equation between the content of o-tyrosine in the electron beam irradiated aquatic products and the dose of electron beam irradiation, and then using high performance liquid chromatography The content of o-tyrosine in aquatic products after electron beam irradiation was detected by the method, and finally the electron beam irradiation dose was obtained through the linear equation. In this technical scheme, phenylalanine in the food reacts with the hydroxyl radicals produced by the rain radiation under the action of radiation to produce tyrosine and two tyrosine isomers, i.e. o-tyrosine (Ortho- Tyrosine, abbreviated as o-tyrosine) and meta-tyrosine (abbreviated as m-tyrosine), tyrosine exists in the body itself, so o-tyrosine and meta-tyrosine are often used as radiation labels It is difficult to separate o-tyrosine and m-tyrosine due to similar polarity, so o-tyrosine is often used in the detection of irradiated food. The invention determines the irradiation dose by detecting the content of o-tyrosine after electron beam irradiation, and provides a theoretical basis for the safety of irradiated food. The detection method of the invention is fast, simple and has high precision.
作为优选，包括以下步骤：Preferably, the following steps are included:
a)原料的选取与预处理：取扇贝、基围虾与鲈鱼进行1-10kGy辐照剂量的电子束辐照处理，匀浆后置于-40℃下保存5-10h；a) Selection and pretreatment of raw materials: scallops, prawns and perch are subjected to electron beam irradiation treatment with an irradiation dose of 1-10kGy, and after homogenization, they are stored at -40°C for 5-10h;
b)前处理：取步骤a)得到的样品1-1.2g，置于具塞离心管中，加入5-6.5mL质量分数为0.2％甲酸水溶液，20000-22000r/min均质4-5min，超声20-35min，10000-12000rpm4℃下离心25-30min，取上清，加入10-15mL丙酮，-40℃放置3-5h，10000-12000rpm4℃下离心8-12min，取上清液于50-55℃悬蒸，除去有机溶剂后冻干，用1-1.2mL质量分数为0.2％甲酸复溶，过0.22μm有机滤膜，上机检测。b) Pretreatment: Take 1-1.2 g of the sample obtained in step a), place it in a centrifuge tube with a stopper, add 5-6.5 mL of 0.2% formic acid aqueous solution, homogenize at 20000-22000 r/min for 4-5 min, and ultrasonically 20-35min, centrifuge at 10000-12000rpm at 4°C for 25-30min, take the supernatant, add 10-15mL acetone, place at -40°C for 3-5h, centrifuge at 10000-12000rpm at 4°C for 8-12min, take the supernatant at 50-55 ℃ suspension steaming, remove the organic solvent and then lyophilize, reconstitute with 1-1.2mL 0.2% formic acid, pass through a 0.22μm organic filter membrane, and test on the machine.
作为优选，高效液相色谱的色谱条件为：色谱柱：AglientEclipse-C18柱(4.6mm×250mm，5.0μm)色谱柱；流动相：A：0.1％甲酸，B：乙醇(20:80，v/v，乙醇中含有体积比为0.5％的冰乙酸和0.3％的三乙胺)；柱温：30℃，波长：Ex:275nm，Em：305nm，进样量：20μL。As preferably, the chromatographic conditions of high performance liquid chromatography are: chromatographic column: AglientEclipse-C18 column (4.6mm * 250mm, 5.0 μ m) chromatographic column; Mobile phase: A: 0.1% formic acid, B: ethanol (20:80, v/ v, ethanol contains 0.5% glacial acetic acid and 0.3% triethylamine by volume); column temperature: 30°C, wavelength: Ex: 275nm, Em: 305nm, injection volume: 20 μL.
作为优选，鲈鱼为Y＝0.1133X3-0.3534X2+5.2943X-1.1651，R2＝0.9986；其中，Y为邻酪氨酸含量，X为电子束辐照剂量。Preferably, the perch is Y=0.1133X 3 -0.3534X 2 +5.2943X-1.1651, R 2 =0.9986; wherein, Y is the content of o-tyrosine, and X is the electron beam irradiation dose.
作为优选，基围虾为Y＝-0.5626X3+10.206X2-4.9731X+6.8588，R2＝0.9938；其中，Y为邻酪氨酸含量，X为电子束辐照剂量。As a preference, Y=-0.5626X 3 +10.206X 2 -4.9731X+6.8588, R 2 =0.9938; wherein, Y is the content of o-tyrosine, and X is the electron beam irradiation dose.
作为优选，扇贝分别为Y＝0.1376X3-2.7062X2+19.949X+2.3847，R2＝0.985；其中，Y为邻酪氨酸含量，X为电子束辐照剂量。Preferably, the scallops are respectively Y=0.1376X 3 -2.7062X 2 +19.949X+2.3847, R 2 =0.985; wherein, Y is the content of o-tyrosine, and X is the electron beam irradiation dose.
作为优选，步骤a)中电子束辐照剂量分别为1kGy，3kGy，5kGy，7kGy，10kGy。Preferably, the electron beam irradiation doses in step a) are 1 kGy, 3 kGy, 5 kGy, 7 kGy, 10 kGy respectively.
本发明的有益效果是本发明通过检测电子束辐照后邻酪氨酸含量的多少来确定辐照剂量，为辐照食品的安全性提供理论基础，本发明的检测方法快速简便、无损、精确度高；检测方法对目标化合物的线性关系好，回收率高，相对偏差小，使用的试剂低毒，使用量少，对环境友好；辐照剂量为1-10kGy的范围内，三种电子束辐照水产品中的邻酪氨酸含量与辐照剂量的关系均可以采用三次函数进行模拟，其相关系数R2＞0.98。The beneficial effect of the present invention is that the present invention determines the irradiation dose by detecting the content of o-tyrosine after electron beam irradiation, and provides a theoretical basis for the safety of irradiated food. The detection method of the present invention is fast, simple, non-destructive and accurate High accuracy; the detection method has a good linear relationship to the target compound, high recovery rate, small relative deviation, low toxicity of reagents, less usage, and environmental friendliness; the irradiation dose is within the range of 1-10kGy, three electron beams The relationship between o-tyrosine content in irradiated aquatic products and irradiation dose can be simulated by cubic function, and the correlation coefficient R 2 >0.98.
附图说明Description of drawings
图1是本发明鲈鱼中邻酪氨酸含量与辐照剂量的线性方程图。Fig. 1 is a linear equation diagram of o-tyrosine content and irradiation dose in perch of the present invention.
图2是本发明基围虾中邻酪氨酸含量与辐照剂量的线性方程图。Fig. 2 is a linear equation diagram of o-tyrosine content and irradiation dose in the shrimp of the present invention.
图3是本发明扇贝中邻酪氨酸含量与辐照剂量的线性方程图。Fig. 3 is a linear equation diagram of o-tyrosine content and irradiation dose in scallops of the present invention.
具体实施方式detailed description
下面通过具体实施方式对本发明做进一步的描述。The present invention will be further described below through specific embodiments.
在本发明中，若非特指，所有设备和原料均可从市场购得或是本行业常用的，下述实施例中的方法，如无特别说明，均为本领域常规方法。In the present invention, unless otherwise specified, all equipment and raw materials can be purchased from the market or commonly used in this industry. The methods in the following examples, unless otherwise specified, are conventional methods in this field.
扇贝、基围虾、鲈鱼购于青岛南山农贸市场；Scallops, prawns and perch were purchased at Nanshan Farmer’s Market in Qingdao;
Agilent1260高效液相色谱仪(配有荧光检测器)购自美国Aglient公司；Agilent1260 high performance liquid chromatograph (equipped with fluorescence detector) is purchased from American Agilent company;
真空旋转蒸发仪(Hei-Vap)购自德国Heidolph公司；Vacuum rotary evaporator (Hei-Vap) was purchased from Heidolph Company, Germany;
高速组织捣碎机(DS-1)购自上海标本模型厂；A high-speed tissue grinder (DS-1) was purchased from Shanghai Specimen Model Factory;
高速分散机购自IKA仪器设备有限公司；The high-speed dispersing machine was purchased from IKA Instrument Equipment Co., Ltd.;
高速冷冻离心机(3K-15)购自德国Sigma公司；A high-speed refrigerated centrifuge (3K-15) was purchased from Sigma, Germany;
超声波清洗器(KQ5200B)购自昆山市超声仪器有限公司；Ultrasonic cleaner (KQ5200B) was purchased from Kunshan Ultrasonic Instrument Co., Ltd.;
电子分析天平(精度0.1g，TE601-2)购自德国赛多利斯有限公司；Electronic analytical balance (precision 0.1g, TE601-2) was purchased from Germany Sartorius Co., Ltd.;
电子分析天平(精度0.0001g，BS-224-S)购自德国赛多利斯有限公司；Electronic analytical balance (accuracy 0.0001g, BS-224-S) was purchased from Germany Sartorius Co., Ltd.;
超纯水系统(Milli-Q)购自美国Millipore公司；The ultrapure water system (Milli-Q) was purchased from Millipore, USA;
磁力搅拌器(GL-3250A)购自北京莱伯泰科学仪器有限公司；The magnetic stirrer (GL-3250A) was purchased from Beijing Labotai Scientific Instrument Co., Ltd.;
AglientEclipse-C18柱(4.6mm×250mm，5.0μm)购自美国Aglient公司；冰乙酸(分析纯)、三乙胺(分析纯)购自国药集团化学试剂有限公司；AglientEclipse-C18 column (4.6mm×250mm, 5.0μm) was purchased from Aglient Company in the United States; Glacial acetic acid (analytical grade) and triethylamine (analytical grade) were purchased from Sinopharm Chemical Reagent Co., Ltd.;
DL-邻酪氨酸(DL-o-tyrosine)购自Sigma公司，纯度≥96.0％。DL-o-tyrosine (DL-o-tyrosine) was purchased from Sigma Company with a purity of ≥96.0%.
高效液相色谱的色谱条件为：色谱柱：AglientEclipse-C18柱(4.6mm×250mm，5.0μm)色谱柱；流动相：A：0.1％甲酸，B：乙醇(20:80，v/v，乙醇中含有体积比为0.5％的冰乙酸和0.3％的三乙胺)；柱温：30℃，波长：Ex:275nm，Em：305nm，进样量：20μL。The chromatographic conditions of high performance liquid chromatography are: chromatographic column: AglientEclipse-C18 column (4.6mm * 250mm, 5.0 μ m) chromatographic column; Mobile phase: A: 0.1% formic acid, B: ethanol (20:80, v/v, ethanol containing 0.5% glacial acetic acid and 0.3% triethylamine in volume ratio); column temperature: 30°C, wavelength: Ex: 275nm, Em: 305nm, injection volume: 20μL.
高效液相色谱梯度洗脱程序：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)。High performance liquid chromatography 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).
实施例1Example 1
一种检测水产品中电子束辐照剂量的方法，包括以下步骤：A method for detecting electron beam radiation dose in aquatic products, comprising the following steps:
a)原料的选取与预处理：取扇贝、基围虾与鲈鱼分别进行1kGy，3kGy，5kGy，7kGy，10kGy辐照剂量的电子束辐照处理，匀浆后置于-40℃下保存5h；a) Selection and pretreatment of raw materials: Scallops, prawns and perch were subjected to electron beam irradiation treatment with irradiation doses of 1kGy, 3kGy, 5kGy, 7kGy, and 10kGy respectively, and the homogenate was stored at -40°C for 5h;
b)前处理：取步骤a)得到的样品1g，置于具塞离心管中，加入5mL质量分数为0.2％甲酸水溶液，20000r/min均质4min，超声20min，10000rpm4℃下离心25min，取上清，加入10mL丙酮，-40℃放置3h，10000rpm4℃下离心8-12min，取上清液于50℃悬蒸，除去有机溶剂后冻干，用1mL质量分数为0.2％甲酸复溶，过0.22μm有机滤膜，上机检测。b) Pretreatment: Take 1 g of the sample obtained in step a), put it in a centrifuge tube with a stopper, add 5 mL of formic acid aqueous solution with a mass fraction of 0.2%, homogenize at 20,000 r/min for 4 min, ultrasonicate for 20 min, and centrifuge at 10,000 rpm for 25 min at 4°C, take the above Add 10mL of acetone, place at -40°C for 3h, centrifuge at 10,000rpm at 4°C for 8-12min, take the supernatant and steam at 50°C, remove the organic solvent and freeze-dry, redissolve in 1mL of 0.2% formic acid, pass through 0.22 μm organic filter membrane, on-board detection.
实施例2Example 2
一种检测水产品中电子束辐照剂量的方法，包括以下步骤：A method for detecting electron beam radiation dose in aquatic products, comprising the following steps:
a)原料的选取与预处理：取扇贝、基围虾与鲈鱼分别进行1kGy，3kGy，5kGy，7kGy，10kGy辐照剂量的电子束辐照处理，匀浆后置于-40℃下保存8h；a) Selection and pretreatment of raw materials: scallops, prawns and perch were subjected to electron beam irradiation treatment with irradiation doses of 1kGy, 3kGy, 5kGy, 7kGy, and 10kGy respectively, and the homogenate was stored at -40°C for 8h;
b)前处理：取步骤a)得到的样品1.1g，置于具塞离心管中，加入5.5mL质量分数为0.2％甲酸水溶液，21000r/min均质4.5min，超声25min，11000rpm4℃下离心28min，取上清，加入12mL丙酮，-40℃放置4h，11000rpm4℃下离心10min，取上清液于53℃悬蒸，除去有机溶剂后冻干，用1.1mL质量分数为0.2％甲酸复溶，过0.22μm有机滤膜，上机检测。b) Pretreatment: Take 1.1 g of the sample obtained in step a), put it in a centrifuge tube with a stopper, add 5.5 mL of 0.2% formic acid aqueous solution, homogenize at 21000 r/min for 4.5 min, ultrasonic for 25 min, and centrifuge at 11000 rpm for 28 min at 4 °C , take the supernatant, add 12mL of acetone, place at -40°C for 4h, centrifuge at 11000rpm at 4°C for 10min, take the supernatant and steam at 53°C, remove the organic solvent and freeze-dry, redissolve with 1.1mL of 0.2% formic acid, Pass through a 0.22μm organic filter membrane and test on the machine.
实施例3Example 3
一种检测水产品中电子束辐照剂量的方法，包括以下步骤：A method for detecting electron beam radiation dose in aquatic products, comprising the following steps:
a)原料的选取与预处理：取扇贝、基围虾与鲈鱼分别进行1kGy，3kGy，5kGy，7kGy，10kGy辐照剂量的电子束辐照处理，匀浆后置于-40℃下保存10h；a) Selection and pretreatment of raw materials: Scallops, prawns and perch were subjected to electron beam irradiation treatment with irradiation doses of 1kGy, 3kGy, 5kGy, 7kGy, and 10kGy respectively, and the homogenate was stored at -40°C for 10h;
b)前处理：取步骤a)得到的样品1.2g，置于具塞离心管中，加入6.5mL质量分数为0.2％甲酸水溶液，22000r/min均质5min，超声35min，12000rpm4℃下离心30min，取上清，加入15mL丙酮，-40℃放置5h，12000rpm4℃下离心12min，取上清液于55℃悬蒸，除去有机溶剂后冻干，用1.2mL质量分数为0.2％甲酸复溶，过0.22μm有机滤膜，上机检测。b) Pre-treatment: take 1.2 g of the sample obtained in step a), place it in a centrifuge tube with a stopper, add 6.5 mL of 0.2% formic acid aqueous solution, homogenize at 22000 r/min for 5 min, sonicate for 35 min, and centrifuge at 12000 rpm for 30 min at 4°C. Take the supernatant, add 15mL of acetone, place it at -40°C for 5h, centrifuge at 12000rpm at 4°C for 12min, take the supernatant and hang steam at 55°C, remove the organic solvent and freeze-dry, redissolve in 1.2mL of 0.2% formic acid, and pass 0.22μm organic filter membrane, on-board detection.
对比例1，取取扇贝、基围虾与鲈鱼不经电子束辐照，其余步骤均与实施例1相同。In comparative example 1, scallops, prawns and perch were not irradiated with electron beams, and the rest of the steps were the same as in Example 1.
对比例1中未检测出邻酪氨酸含量。No o-tyrosine content was detected in Comparative Example 1.
表1为实施例1-3中不同剂量下鲈鱼中的邻酪氨酸含量。Table 1 is the content of o-tyrosine in perch under different doses in Examples 1-3.
表1、不同剂量下鲈鱼中的邻酪氨酸含量Table 1. O-tyrosine content in sea bass under different doses
鲈鱼中邻酪氨酸含量与辐照剂量关系图见图1。The relationship between o-tyrosine content and irradiation dose in perch is shown in Figure 1.
鲈鱼为Y＝0.1133X3-0.3534X2+5.2943X-1.1651，R2＝0.9986；其中，Y为邻酪氨酸含量，X为电子束辐照剂量。Perch is Y=0.1133X 3 -0.3534X 2 +5.2943X-1.1651, R 2 =0.9986; wherein, Y is the content of o-tyrosine, and X is the dose of electron beam irradiation.
表2为实施例1-3中不同剂量下基围虾中的邻酪氨酸含量。Table 2 is the content of ortho-tyrosine in the shrimp under different doses in Examples 1-3.
表2、不同辐照剂量下基围虾中的邻酪氨酸含量Table 2. The o-tyrosine content in the base shrimp under different irradiation doses
基围虾中的邻酪氨酸含量与辐照剂量关系图见图2。See Figure 2 for the relationship between o-tyrosine content and irradiation dose in genotype shrimp.
基围虾为Y＝-0.5626X3+10.206X2-4.9731X+6.8588，R2＝0.9938；其中，Y为邻酪氨酸含量，X为电子束辐照剂量。The basal shrimp is Y=-0.5626X 3 +10.206X 2 -4.9731X+6.8588, R 2 =0.9938; wherein, Y is the content of o-tyrosine, and X is the electron beam irradiation dose.
表3为实施例1-3中不同剂量下扇贝中的邻酪氨酸含量。Table 3 shows the o-tyrosine content in scallops with different doses in Examples 1-3.
表3、不同辐照剂量下扇贝中的邻酪氨酸含量Table 3. O-tyrosine content in scallops under different irradiation doses
扇贝中中的邻酪氨酸含量与辐照剂量关系图见图3。The relationship between o-tyrosine content and irradiation dose in scallops is shown in Figure 3.
扇贝分别为Y＝0.1376X3-2.7062X2+19.949X+2.3847，R2＝0.985；其中，Y为邻酪氨酸含量，X为电子束辐照剂量。Scallops are respectively Y=0.1376X 3 -2.7062X 2 +19.949X+2.3847, R 2 =0.985; wherein, Y is the content of o-tyrosine, and X is the electron beam irradiation dose.
从表1-3可知三种电子束辐照水产品中的邻酪氨酸含量随着辐照剂量的升高而增大，不同种类水产品中的邻酪氨酸含量不同，与水产品自身的结构和性质相关；辐照剂量为1-10kGy的范围内，三种电子束辐照水产品中的邻酪氨酸含量与辐照剂量的关系均可以采用三次函数进行模拟，见图1-3，其相关系数R2＞0.98。It can be seen from Table 1-3 that the content of o-tyrosine in the three types of electron beam irradiated aquatic products increases with the increase of the irradiation dose, and the content of o-tyrosine in different types of aquatic products is different, which is different from that of the aquatic products themselves. In the range of 1-10kGy irradiation dose, the relationship between o-tyrosine content and irradiation dose in three kinds of electron beam irradiated aquatic products can be simulated by cubic function, as shown in Figure 1- 3. The correlation coefficient R 2 >0.98.
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