本文選題：時(shí)間溫度指示器 + 大菱鲆生魚(yú)片��； 參考：《中國(guó)海洋大學(xué)》2014年碩士論文

【摘要】：溫度對(duì)食品在儲(chǔ)藏和運(yùn)輸過(guò)程中的質(zhì)量變化有著十分重要的影響。然而目前，我國(guó)以時(shí)間為基礎(chǔ)的食品標(biāo)簽制度雖然可以提供指示食品腐敗程度的保質(zhì)期，但是由于其無(wú)法直接反映食品在實(shí)際儲(chǔ)存和運(yùn)輸過(guò)程中所經(jīng)歷的溫度歷史，所以難以精確指示食品的新鮮程度。而對(duì)于一些直接生食的高端水產(chǎn)品或者鮮度要求極高的食品而言，簡(jiǎn)單準(zhǔn)確判斷食品鮮度十分重要。因此需要一種有效而直觀的方式同時(shí)指示食品所經(jīng)歷的溫度和時(shí)間。時(shí)間溫度指示器(Time temperatureindicator, TTI)是一種簡(jiǎn)單方便、經(jīng)濟(jì)有效的指示時(shí)間溫度的方法。然而，目前我國(guó)對(duì)于TTI的研究尚處于起步階段，理論研究不系統(tǒng)，開(kāi)發(fā)的TTI產(chǎn)品因存在諸多問(wèn)題而鮮有應(yīng)用。 本論文以酪氨酸和酪氨酸酶為研究對(duì)象，利用酶促反應(yīng)原理，開(kāi)發(fā)一種新型固態(tài)酶型TTI。經(jīng)可行性分析、指示時(shí)間優(yōu)化、動(dòng)力學(xué)分析等系統(tǒng)的理論研究后，設(shè)計(jì)了一種可以指示食品鮮度的TTI產(chǎn)品，并將其初步應(yīng)用于大菱鲆生魚(yú)片的質(zhì)量指示。主要研究?jī)?nèi)容如下： 1、分析了酪氨酸酶與酪氨酸液相反應(yīng)體系的褐變趨勢(shì)，發(fā)現(xiàn)不同溫度和時(shí)間條件下酶促反應(yīng)體系的褐變程度有著顯著的不同，闡明了開(kāi)發(fā)以酪氨酸酶促反應(yīng)為基礎(chǔ)的TTI的可行性。為進(jìn)一步證實(shí)開(kāi)發(fā)固態(tài)TTI產(chǎn)品的可行性，研究了固定化底物釋放速率的單因素實(shí)驗(yàn)，結(jié)果表明：酪氨酸在固定化后可以從聚乙烯醇凝膠薄膜中緩慢釋放并與酪氨酸酶進(jìn)行反應(yīng)，且在聚乙烯醇濃度為15%，酪氨酸含量為0.6mg/mL，反復(fù)冷凍-解凍循環(huán)法成膜的條件下，固相體系酪氨酸的釋放速率最低，體系達(dá)到反應(yīng)終點(diǎn)的時(shí)間約為18h。 2、將酪氨酸和酪氨酸酶分別固定化于聚乙烯醇凝膠緩釋薄膜，開(kāi)發(fā)新型的以酪氨酸酶促褐變?yōu)榛A(chǔ)的固態(tài)酶型時(shí)間溫度指示器，并根據(jù)指示時(shí)間優(yōu)化其制備條件。采用總體顏色變化ΔE來(lái)判斷TTI指示時(shí)間，選取ΔE值不再有顯著變化時(shí)的值（ΔE=38.21±2.00左右）作為T(mén)TI的指示終點(diǎn)。通過(guò)響應(yīng)面實(shí)驗(yàn)，成功建立了以TTI指示時(shí)間為響應(yīng)值，以酪氨酸含量、酪氨酸酶含量以及聚乙烯醇濃度為自變量的數(shù)學(xué)模型，其公式為：t=50.40+0.25A+0.50B+1.50C-10.33A2-7.83B2-8.83C2-3.75AB+0.75AC+1.25BC，其中t為指示時(shí)間，A為酪氨酸含量，B為酪氨酸酶含量，C為聚乙烯醇濃度。根據(jù)模型預(yù)測(cè)，確定了TTI達(dá)到最長(zhǎng)指示時(shí)間時(shí)各自變量的取值：酪氨酸30.08mg，，酪氨酸酶0.30mL，聚乙烯醇15.18%，此時(shí)TTI的最長(zhǎng)指示時(shí)間可以達(dá)到50.48h。 3、選取響應(yīng)面實(shí)驗(yàn)優(yōu)化的TTI制備條件，制備了一種新型固態(tài)酶型TTI，并對(duì)其進(jìn)行動(dòng)力學(xué)分析，為指示產(chǎn)品的選擇及TTI的應(yīng)用提供理論基礎(chǔ)。結(jié)果表明該TTI的反應(yīng)活化能Ea=0.409kJ/mol，指前因子A=2.91。采用SAS軟件對(duì)TTI進(jìn)行數(shù)學(xué)模型的分析，結(jié)果表明，時(shí)間t、溫度T和顏色響應(yīng)值F(X)之間滿(mǎn)足一個(gè)二元二次函數(shù)關(guān)系式F(X)=-0.00586T2-0.24002T+0.00902T×t-0.02060t-1.73930，該式中R2=0.9661，說(shuō)明模型擬合度良好。此模型建立了該TTI體系顏色變化與所處時(shí)間溫度之間的關(guān)系，為指示器的應(yīng)用打下基礎(chǔ)。 4、選取了鮮度要求高且附加值高的大菱鲆生魚(yú)片作為T(mén)TI指示的產(chǎn)品，首先研究了其鮮度變化。通過(guò)測(cè)定大菱鲆從初始至死亡48h內(nèi)，不同溫度(4、12、18℃)下大菱鲆魚(yú)肉中菌落總數(shù)、TVBN、pH和失重率變化情況，闡明了大菱鲆生魚(yú)片從0h至48h的鮮度變化情況。實(shí)驗(yàn)結(jié)果表明，大菱鲆生魚(yú)片菌落總數(shù)在48h內(nèi)變化顯著，且4℃下儲(chǔ)藏48h左右的大菱鲆生魚(yú)片已超出生食標(biāo)準(zhǔn)；而TVBN、pH和失重率在48h內(nèi)則無(wú)明顯變化。因此，選用菌落總數(shù)表征大菱鲆生魚(yú)片的鮮度指標(biāo)。通過(guò)對(duì)大菱鲆生魚(yú)片鮮度變化的動(dòng)力學(xué)分析，計(jì)算鮮度活化能為0.009kJ/mol，這與之前計(jì)算的TTI的活化能（0.409kJ/mol）接近，符合TTI對(duì)應(yīng)產(chǎn)品的標(biāo)準(zhǔn)（產(chǎn)品活化能與TTI活化能相差小于40kJ/mol）。這表明本實(shí)驗(yàn)中設(shè)計(jì)的TTI可以很好地應(yīng)用于大菱鲆生魚(yú)片的鮮度指示。實(shí)驗(yàn)中同時(shí)設(shè)計(jì)了TTI結(jié)構(gòu)，該結(jié)構(gòu)可以粘附到生魚(yú)片產(chǎn)品外包裝表面，易于激活，保護(hù)性良好，結(jié)構(gòu)小巧，且透明材質(zhì)的設(shè)計(jì)，方便消費(fèi)者觀察。 研究表明開(kāi)發(fā)出的新型固態(tài)酶型TTI，反應(yīng)體系簡(jiǎn)單，顏色變化明顯，可以用于判斷極易腐敗低溫流通食品（如大菱鲆生魚(yú)片）的新鮮程度，為生魚(yú)片供應(yīng)鏈中的制造商、流通商以及零售商提供了一種有效地產(chǎn)品質(zhì)量控制手段，同時(shí)對(duì)于消費(fèi)者更準(zhǔn)確地判定此類(lèi)食品是否適于生食有著積極的意義。
[Abstract]:Temperature has a very important impact on the quality of food in storage and transportation. However, the time based food labeling system in our country can provide a shelf life indicating the degree of food corruption, but it does not directly reflect the temperature history of the food in the actual storage and transportation process. It is difficult to accurately indicate the freshness of the food. It is important to determine the food freshness simply and accurately for some high end aquatic products with direct food or high freshness. Therefore, an effective and intuitive way is needed to indicate the temperature and time of the food at the same time. The time temperature indicator (Time temperaturein) Dicator, TTI) is a simple, convenient, economical and effective method to indicate time and temperature. However, at present, the research on TTI is still in its infancy, the theoretical research is not systematic, and the developed TTI products are rarely applied because of many problems.
In this paper, using tyrosine and tyrosinase as the research object, using the principle of enzymatic reaction, we developed a new solid-state enzyme type TTI. through the feasibility analysis, indicating time optimization, dynamic analysis and other systems. A kind of TTI product which can indicate food freshness was designed and applied to the quality of turbot sashimi slices. The main contents of the study are as follows:
1, the browning trend of tyrosinase and tyrosine liquid phase reaction system was analyzed. It was found that the browning degree of the enzyme catalyzed reaction system was significantly different under different temperatures and time conditions. The feasibility of developing TTI based on tyrosinase catalyzed reaction was clarified. The immobilization was studied to further confirm the feasibility of developing solid TTI products. The single factor experiment of the rate of substrate release shows that tyrosine can be released slowly and react with tyrosinase after immobilization, and the release rate of tyrosine in solid phase system is the most in the condition of the concentration of polyvinyl alcohol 15%, the tyrosine content is 0.6mg/mL, and the repeated freezing thawing cycle method is used. Low, the time for the system to reach the end of the reaction is about 18h.
2, tyrosine and tyrosinase were immobilized on the slow-release film of polyvinyl alcohol gel, and a new type of solid-state temperature indicator was developed based on tyrosinase promoting browning, and the preparation conditions were optimized according to the indication time. The TTI indication time was determined by the overall color change Delta E, and the value of delta E was no longer significantly changed. The value (delta E=38.21 + 2) is the terminal point of the TTI. Through the response surface experiment, a mathematical model with the content of tyrosine, tyrosinase and the concentration of polyvinyl alcohol as the independent variable is established by the response surface experiment. The formula is: t=50.40+0.25A+0.50B+ 1.50C-10.33A2-7.83B2-8.83C2-3.75AB+0.75AC+1.25BC, of which t For the indication time, A is tyrosine content, B is tyrosinase content and C is polyvinyl alcohol concentration. According to the model prediction, the values of each variable are determined when TTI reaches the longest indication time: tyrosine 30.08mg, tyrosinase 0.30mL, polyvinyl alcohol 15.18%, and at this time the longest time of TTI can reach 50.48h.
3, a new solid-state enzyme type TTI was prepared by selecting the TTI preparation conditions optimized by the response surface experiment. The kinetic analysis was carried out to provide a theoretical basis for the selection of the product and the application of TTI. The results showed that the activation energy of the TTI was Ea=0.409kJ/mol and the former factor A=2.91. was analyzed by SAS software for the mathematical model of TTI. It shows that the time t, the temperature T and the color response value F (X) satisfy a two element two function relation F (X) =-0.00586T2-0.24002T+0.00902T x t-0.02060t-1.73930, which indicates that the model has good fitting degree. This model establishes the relationship between the color change of the TTI system and the temperature of the time, and lays the foundation for the application of the indicator. Foundation.
4, the freshness of turbot sashimi slices with high freshness and high added value was selected as a TTI indicator. First, the freshness of turbot was studied. The total number of colonies, TVBN, pH and weight loss of turbot turbot in turbot turbot from initial to death 48h and at different temperatures (4,12,18 degrees C) were determined. The fresh fish slices from 0h to 48h were clarified. The results showed that the total number of the total colony of turbot sashimi slices changed significantly in 48h, and the sashimi slices of turbot turbot, which was stored for about 48h at 4 C, had exceeded the standard of birth food, while the TVBN, pH and weightlessness rate had no obvious changes in 48h. The kinetic analysis of the fresh degree change shows that the activation energy of the freshness is 0.009kJ/mol, which is close to the activation energy (0.409kJ/mol) of the previously calculated TTI, which is in line with the standard of the corresponding product of the TTI (the difference between the activation energy of the product and the activation energy of TTI is less than 40kJ/mol). This shows that the designed TTI can be well applied to the freshness indication of the sashimi slices of turbot. The TTI structure was designed at the same time. The structure can be attached to the surface of the sashimi product on the outside of the packaging. It is easy to activate, with good protection, small structure, and a transparent material design, which is convenient for the consumer to observe.
The research shows that the new solid state enzyme type TTI has a simple reaction system and the color change is obvious. It can be used to judge the freshness of the highly corrupt low temperature circulation food (such as the turbot sashimi). It provides an effective means of quality control for the manufacturers, tradable merchants and retailers in the supply chain of sashimi. It is of great significance to judge more accurately whether this kind of food is suitable for raw food.

【學(xué)位授予單位】：中國(guó)海洋大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類(lèi)號(hào)】：TS254.7

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