冰温保鲜技术对牛肉品质的影响研究
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摘要
冰温贮藏是将生鲜食品贮藏在0℃到其冰点以上的温度范围内。这样既使生鲜食品生理活性维持在最低程度,又能维持其正常的新陈代谢,抑制微生物的生长,而且不产生冻害,使食品能够长期保鲜。冰温技术以其卓越的贮藏的优越性,在美、日、韩等国得到了长足的发展和应用。近年来,国内对冰温的研究也逐渐增多,在冰温下贮藏蓝莓、菠菜、梨、南美白对虾、鸡肉等的研究都取得了一定的成效。但目前冰温贮藏技术的研究主要还是针对果品蔬菜和水产品,对肉制品,特别的牛肉的研究的相对较少。本研究主要以牛肉为原料,将冰温技术与真空包装技术、保鲜剂保鲜技术、气调保鲜技术相结合,分别对牛肉进行贮藏,并测定贮藏期间牛肉的理化性质、结构特性等,并利用电子鼻技术对牛肉的贮藏货架期进行预测,取得了良好的效果。主要结论如下：
(1)选取不同地区不同部位的牛肉进行冰点测定,研究冰点与水分含量和可溶性固形物含量之间的关系。结果表明,不同地区不同牛胴体部位的牛肉的冰点、含水量以及可溶性固形物之间均存在一定的差异性,牛胴体上位置比较接近的牛肉的含水量和可溶性之间比较接近；不同胴体部位对冰点、含水量以及可溶性固形物的影响要大于不同地区对三个参数的影响；相关性分析发现,水分含量与冰点温度值之间存在假性相关性,所以,不能单单通过水分含量来预测冰点温度值；可溶性固形物的含量与冰点温度值之间有很强的负相关性；对冰点温度和可溶性固形物之间进行回归分析,得到一元线性方程Y=-0.098X-0.208,经验证此方程可以合理地预测不同地区不同胴体部位牛肉的冰点温度值。
(2)以冰温+真空(Cv)、0℃+真空(Zv)、4℃+真空(Fv)、0℃(Z)分别与真空包装相结合,研究贮藏期间牛肉品质指标的变化情况。综合贮藏期间菌落总数和挥发性盐基氮值发现,4种条件下牛肉的货架期分别为32天、24天,20天和8天。相同温度下,菌落总数、TVB-N值以及pH值的变化速度均是Zv (3)对Cv、Zv、Fv这3种贮藏条件下的牛肉进行了从宏观组织质构到微观结构变化的系统研究。除了肌节长度外,其他指标(TPA参数、蛋白质溶解度、巯基含量等)与贮藏时间之间都存在显著或极显著的相关性(p<0.05或p<0.01),说明这些指标对贮藏货架期都有一定的指示作用。对结构指标的测定显示,指标的变化基本上是温度越高,指标变化越快,所以温度对贮藏货架期的影响大。3种贮藏条件下,通过对指标的观测,Cv条件最能维持结构的稳定性,其次是Zv条件,较差的是Fv条件。
(4)对牛肉的TPA参数(硬度、凝聚性、弹性、胶性、咀嚼性)和嫩度的测定显示,随着贮藏时间的延长,凝聚性和嫩度先上升再下降,弹性存在一定的波动性,硬度、胶性和咀嚼性则不断下降。相关分析表明,嫩度与TPA指标之间显著相关。利用共聚焦显微镜对牛肉的肌节长度和肌纤维直径进行测定,结果显示,肌节长度经历了先下降再上升的过程,而肌纤维直径则因失水而逐渐变小。由于内源酶以及外源微生物等作用,蛋白质不断降解,从而导致MFI在贮藏过程中不断增大。MFI可以反映组织结构的降解程度,相关性分析发现,其与同样能够反映降解程度的嫩度指标之间存在着极显著的相关性(p<0.01),可以用MFI来衡量牛肉的嫩度。
(5)贮藏期间蛋白质的溶解度不断下降,溶解度的变化是由蛋白质的变性引起的。而巯基的变化反映了蛋白质的氧化程度。贮藏过程中巯基不断氧化形成二硫键,其含量不断下降。蛋白质溶解度与巯基含量之间存在极显著的相关性(p<0.01)。通过傅里叶变换红外光谱法对提取的可溶性蛋白中的二级结构含量的变化做了进一步的分析,结果表明贮藏末期蛋白质二级结构中的α-螺旋含量减少,而β-折叠片、β-转角与无规则卷曲的含量都出现了一定的升高,这可能是因为蛋白质氧化遭到自由基的攻击,自由基争夺氢离子,导致氢键的断裂,从而α-螺旋结构被破坏,向无规则卷曲形式转变。
(6)利用保鲜剂壳聚糖、茶多酚和Nisin与冰温结合贮藏牛肉。结果发现,3者的保鲜效果依次为壳聚糖>茶多酚>Nisin,三种保鲜剂之间存在显著的交互作用在。冰温条件下,复合保鲜剂的最佳复合条件为：壳聚糖1.70%,Nisin0.03%,茶多酚0.24%。通过Mintab15软件建立回归模型,回归方程为Y=12.55-1.08A-0.70B-1.08C+0.99A2+0.73B2+0.52C2+0.38AB+0.25AC+0.26BC,模型验证误差为2.28%,可以较好地反映在冰温结合复合保鲜剂对牛肉的保鲜效果。与单纯冰温贮藏相比,冰温与复合保鲜剂的结合可以有效延长牛肉的货架期。
(7)冰温与气调包装相结合对牛肉的研究发现,与单纯冰温贮藏相比,冰温与气调结合可以将牛肉的货架期延长4天以上,50%02+50%C02组和30%02+70%C02组的保鲜效果要优于80%02+20%C02组。冰温结合气调包装组牛肉的TVB-N值远远小于冰温对照组,几组气调包装间无显著性差异。高氧气调包装会加大牛肉的失水率。高的氧气含量使鲜肉有较好的色泽,但是贮藏后期肉样表面有褐变产生,较高的二氧化碳对颜色也会产生不利的影响。冰温结合50%O2+50%CO2可以更好地维护肉的红度。
(8)用电子鼻对冰温和普通冷藏条件下牛肉新鲜度进行预测。电子鼻可以很好地区分开不同贮藏时间的牛肉。电子鼻以菌落总数、TVB-N、感官评价值建立模型,并用模型对未参与建模的牛肉进行指标预测。预测值与实测值的回归分析表明,模型可以很好地预测各自所代表的贮藏条件下的牛肉的TVB-N值、菌落总数、感官评分值。电子鼻模型的建立为不同低温贮藏条件下牛肉品质的测定提供了有效而快捷的方法,对牛肉低温贮藏期间的品质监控意义深远。
Controlled freezing point (CFP) storage refers to the storage of fresh food within the temperature range below0℃but above the freezing-point, which can maintain physiological activity of fresh food to the lowest degree, also maintain its normal metabolism, inhibit the growth of microorganisms and cause no cold damage, thus achieve the effect of maintaining long-time freshness. In recent years, CFP technology has been widely applied in the US, Japan and South Korea, etc, and domestic studies on CFP have also gradually increased. Certain achievements have been made on the storage of blueberry, pears, spinach, Penaeus vannamei and chicken at CFP. But present CFP storage studies are mainly related to fruits, vegetables and aquatic products; CFP storage studies on the chilled meat, particularly the beef, are relatively fewer. This study mainly deals with the beef stored at CFP, combined with vacuum conditions, preservatives, modified atmosphere packaging separately as the storage methods for retaining freshness, the physicochemical properties and structural characteristics of the beef during storage were measured, electronic nose technology was used for the beef storage shelf life prediction, which proved good results. The main conclusions are as follows:
(1) Beef of different parts and from different regions was selected for testing at freezing point to determine the relationship between the freezing point and moisture content&the soluble solids. The results showed that beef of different carcass parts and from different regions varied in terms of freezing point, moisture content and soluble solids, the moisture content and soluble solids of the beef from the adjacent parts on the carcass were relatively similar; different parts of the carcass had larger effect on the three parameters of freezing point, moisture content and soluble solids than that of different regions; the correlation analysis indicated that moisture content was spuriously correlated with the temperature value of freezing point, therefore the moisture content alone can not be used for prediction of freezing temperature value; there was a strong negative correlation between soluble solids content and freezing temperature value; the regression analysis on the relationship between the freezing temperature and soluble solids resulted in a unary linear equation Y=-0.098X-0.208, which was verified to be capable of giving reasonable prediction of temperature values of freezing point for the beef of different carcass parts and from different regions.
(2) The storage methods of CFP+vacuum (Cv),0℃+vacuum (Zv),4℃+vacuum (Fv),0℃(Z) were combined with vacuum-packing respectively to investigate the change of beef quality indicators during storage. Given the total plate count and total volatile basic nitrogen during storage, the shelf life of beef under four conditions was32days,24days,20days and8days. The change rate of total plate count, TVB-N, and pH at the same temperature was Zv (3) A systematic study in relation to the macroscopic to microscopic structure change of the chilled beef under the three storage conditions Cv, Zv, Fv was conducted., the other indicators (TPA parameters, tenderness, muscle fiber diameter, protein solubility, myofibrillar fragmentation index, sulfhydryl content, secondary structure content), except the sarcomere length, had significant or highly significant correlation with the storage time (p<0.05or p <0.01), signifying these indicators were of a certain indication to the storage shelf life. The structural indicators show the change of indicators is such that the higher the temperature is, the faster the indicator changes, so the temperature has a large impact on the storage shelf life. For the above three kinds of storage conditions, based on the indicators observed, the structure stability under Cv condition is best maintained, followed by Zv condition and Fv condition comes last.
(4) The measured TPA parameters (hardness, cohesiveness, elasticity, colloidality, chewiness) and tenderness of the beef showed that with the extension of storage time, the cohesiveness and tenderness first increased and then decreased, for elasticity, there was a certain degree of fluctuation, while hardness, colloidality and chewiness were declining. The correlation analysis revealed there was a significant correlation between tenderness and TPA parameters. The beef muscle sarcomere length and radius measured with confocal microscopy showed the sarcomere length first decreased and then increased, while the muscle fiber radius beccame smaller due to dehydration. Because of action of endogenous and exogenous microbial enzymes, the protein was degraded constantly, resulting in the increase in MFI during storage. MFI can reflect the degradation degree of the tissue structure, and the correlation analysis indicates that it has a highly significant correlation (p<0.01) with tenderness indicator that can also reflect the degree of degradation, as a result, MFI can be used to measure the MFI beef tenderness.
(5) The solubility of the protein during storage was declining caused by the denaturation of the protein. The change of thiol group reflects the degree of oxidation of the protein; the thiol was continuously oxidized during storage to become disulfide bond, with the content decreasing. Protein solubility is significantly correlated with thiol levels (p<0.01). Fourier transform infrared spectroscopy was applied for further analysis of secondary structure content change of the soluble protein extracted, and the result showed that a-helix content of protein secondary structure decreased at the end of storage, while the content of β-pleated sheet, β-turn and random coil increased at a certain degree, was a certain, which was probably due to the protein oxidation attacked by free radical and free radical capturing the hydrogen ion, leading to breakage of hydrogen bonds, and a-helix structure was damaged and changed toward the random coil.
(6) The preservative chitosan, polyphenols and Nisin combined with CFP for storage of beef. It was found that the preservation effect of these three methods was chitosan> TP> Nisin, and there existed significant interaction between these three kinds of preservatives. Under CFP condition, the optimum compound conditions for compound preservative are: chitosan1.70%, Nisin0.03%, polyphenols0.24%. A regression model was build on Mintab15software, resulting in the regression equation of Y=12.55-1.08A-0.70B-1.08C+0.99A2+0.73B2+0.52C2+0.38AB+0.25AC+0.26BC, with model validation error of2.28%, which can better reflect the preservation effect of the compound preservative combined with CFP storage on beef. Compared with pure CFP storage, the combination of CFP storage with compound preservative combination can effectively extend the shelf life of chilled beef.
(7) The study on beef CFP storage combined with modified atmosphere packaging found that compared with the CFP storage alone, the combination of CFP storage with modified atmosphere packaging can extend the shelf life of beef for over4days, the preservation effect of50%02+50%CO2group and30%02+70%CO2group was better than that of80%02+20%CO2group. The beef TVB-N under CFP storage combined with modified atmosphere packaging was far less than the control group of CFP storage, no significant difference was found between several groups of modified atmosphere packaging. High oxygen modified atmosphere packaging will increase the rate of water loss of beef. High oxygen content gained better meat color, yet causing brown stain on the beef surface in later period of the storage, and the higher the carbon dioxide will also adversely affect the color. CFP storage combined with50%O2+50%CO2can better maintain the meat freshness.
(8) The electronic nose was used for freshness prediction of the beef stored at CFP and under normal storage. Electronic nose can well distinguish the beef of different storage times. The electronic nose builds models based on parameters of total plate count, TVB-N and sensory evaluation value, which can be used for indicator forecast of the beef not involved in modeling. The regression analysis of the predicted and measured values show the model can well predict total plate count, TVB-N and sensory evaluation value of the beef under the corresponding storage conditions. The establishment of an electronic nose model provides an effective and efficient method for determining the quality of chilled beef under different storage conditions, which is of great significance for quality control of chilled fresh beef during storage.
(1)选取不同地区不同部位的牛肉进行冰点测定,研究冰点与水分含量和可溶性固形物含量之间的关系。结果表明,不同地区不同牛胴体部位的牛肉的冰点、含水量以及可溶性固形物之间均存在一定的差异性,牛胴体上位置比较接近的牛肉的含水量和可溶性之间比较接近；不同胴体部位对冰点、含水量以及可溶性固形物的影响要大于不同地区对三个参数的影响；相关性分析发现,水分含量与冰点温度值之间存在假性相关性,所以,不能单单通过水分含量来预测冰点温度值；可溶性固形物的含量与冰点温度值之间有很强的负相关性；对冰点温度和可溶性固形物之间进行回归分析,得到一元线性方程Y=-0.098X-0.208,经验证此方程可以合理地预测不同地区不同胴体部位牛肉的冰点温度值。
(2)以冰温+真空(Cv)、0℃+真空(Zv)、4℃+真空(Fv)、0℃(Z)分别与真空包装相结合,研究贮藏期间牛肉品质指标的变化情况。综合贮藏期间菌落总数和挥发性盐基氮值发现,4种条件下牛肉的货架期分别为32天、24天,20天和8天。相同温度下,菌落总数、TVB-N值以及pH值的变化速度均是Zv
(4)对牛肉的TPA参数(硬度、凝聚性、弹性、胶性、咀嚼性)和嫩度的测定显示,随着贮藏时间的延长,凝聚性和嫩度先上升再下降,弹性存在一定的波动性,硬度、胶性和咀嚼性则不断下降。相关分析表明,嫩度与TPA指标之间显著相关。利用共聚焦显微镜对牛肉的肌节长度和肌纤维直径进行测定,结果显示,肌节长度经历了先下降再上升的过程,而肌纤维直径则因失水而逐渐变小。由于内源酶以及外源微生物等作用,蛋白质不断降解,从而导致MFI在贮藏过程中不断增大。MFI可以反映组织结构的降解程度,相关性分析发现,其与同样能够反映降解程度的嫩度指标之间存在着极显著的相关性(p<0.01),可以用MFI来衡量牛肉的嫩度。
(5)贮藏期间蛋白质的溶解度不断下降,溶解度的变化是由蛋白质的变性引起的。而巯基的变化反映了蛋白质的氧化程度。贮藏过程中巯基不断氧化形成二硫键,其含量不断下降。蛋白质溶解度与巯基含量之间存在极显著的相关性(p<0.01)。通过傅里叶变换红外光谱法对提取的可溶性蛋白中的二级结构含量的变化做了进一步的分析,结果表明贮藏末期蛋白质二级结构中的α-螺旋含量减少,而β-折叠片、β-转角与无规则卷曲的含量都出现了一定的升高,这可能是因为蛋白质氧化遭到自由基的攻击,自由基争夺氢离子,导致氢键的断裂,从而α-螺旋结构被破坏,向无规则卷曲形式转变。
(6)利用保鲜剂壳聚糖、茶多酚和Nisin与冰温结合贮藏牛肉。结果发现,3者的保鲜效果依次为壳聚糖>茶多酚>Nisin,三种保鲜剂之间存在显著的交互作用在。冰温条件下,复合保鲜剂的最佳复合条件为：壳聚糖1.70%,Nisin0.03%,茶多酚0.24%。通过Mintab15软件建立回归模型,回归方程为Y=12.55-1.08A-0.70B-1.08C+0.99A2+0.73B2+0.52C2+0.38AB+0.25AC+0.26BC,模型验证误差为2.28%,可以较好地反映在冰温结合复合保鲜剂对牛肉的保鲜效果。与单纯冰温贮藏相比,冰温与复合保鲜剂的结合可以有效延长牛肉的货架期。
(7)冰温与气调包装相结合对牛肉的研究发现,与单纯冰温贮藏相比,冰温与气调结合可以将牛肉的货架期延长4天以上,50%02+50%C02组和30%02+70%C02组的保鲜效果要优于80%02+20%C02组。冰温结合气调包装组牛肉的TVB-N值远远小于冰温对照组,几组气调包装间无显著性差异。高氧气调包装会加大牛肉的失水率。高的氧气含量使鲜肉有较好的色泽,但是贮藏后期肉样表面有褐变产生,较高的二氧化碳对颜色也会产生不利的影响。冰温结合50%O2+50%CO2可以更好地维护肉的红度。
(8)用电子鼻对冰温和普通冷藏条件下牛肉新鲜度进行预测。电子鼻可以很好地区分开不同贮藏时间的牛肉。电子鼻以菌落总数、TVB-N、感官评价值建立模型,并用模型对未参与建模的牛肉进行指标预测。预测值与实测值的回归分析表明,模型可以很好地预测各自所代表的贮藏条件下的牛肉的TVB-N值、菌落总数、感官评分值。电子鼻模型的建立为不同低温贮藏条件下牛肉品质的测定提供了有效而快捷的方法,对牛肉低温贮藏期间的品质监控意义深远。
Controlled freezing point (CFP) storage refers to the storage of fresh food within the temperature range below0℃but above the freezing-point, which can maintain physiological activity of fresh food to the lowest degree, also maintain its normal metabolism, inhibit the growth of microorganisms and cause no cold damage, thus achieve the effect of maintaining long-time freshness. In recent years, CFP technology has been widely applied in the US, Japan and South Korea, etc, and domestic studies on CFP have also gradually increased. Certain achievements have been made on the storage of blueberry, pears, spinach, Penaeus vannamei and chicken at CFP. But present CFP storage studies are mainly related to fruits, vegetables and aquatic products; CFP storage studies on the chilled meat, particularly the beef, are relatively fewer. This study mainly deals with the beef stored at CFP, combined with vacuum conditions, preservatives, modified atmosphere packaging separately as the storage methods for retaining freshness, the physicochemical properties and structural characteristics of the beef during storage were measured, electronic nose technology was used for the beef storage shelf life prediction, which proved good results. The main conclusions are as follows:
(1) Beef of different parts and from different regions was selected for testing at freezing point to determine the relationship between the freezing point and moisture content&the soluble solids. The results showed that beef of different carcass parts and from different regions varied in terms of freezing point, moisture content and soluble solids, the moisture content and soluble solids of the beef from the adjacent parts on the carcass were relatively similar; different parts of the carcass had larger effect on the three parameters of freezing point, moisture content and soluble solids than that of different regions; the correlation analysis indicated that moisture content was spuriously correlated with the temperature value of freezing point, therefore the moisture content alone can not be used for prediction of freezing temperature value; there was a strong negative correlation between soluble solids content and freezing temperature value; the regression analysis on the relationship between the freezing temperature and soluble solids resulted in a unary linear equation Y=-0.098X-0.208, which was verified to be capable of giving reasonable prediction of temperature values of freezing point for the beef of different carcass parts and from different regions.
(2) The storage methods of CFP+vacuum (Cv),0℃+vacuum (Zv),4℃+vacuum (Fv),0℃(Z) were combined with vacuum-packing respectively to investigate the change of beef quality indicators during storage. Given the total plate count and total volatile basic nitrogen during storage, the shelf life of beef under four conditions was32days,24days,20days and8days. The change rate of total plate count, TVB-N, and pH at the same temperature was Zv
(4) The measured TPA parameters (hardness, cohesiveness, elasticity, colloidality, chewiness) and tenderness of the beef showed that with the extension of storage time, the cohesiveness and tenderness first increased and then decreased, for elasticity, there was a certain degree of fluctuation, while hardness, colloidality and chewiness were declining. The correlation analysis revealed there was a significant correlation between tenderness and TPA parameters. The beef muscle sarcomere length and radius measured with confocal microscopy showed the sarcomere length first decreased and then increased, while the muscle fiber radius beccame smaller due to dehydration. Because of action of endogenous and exogenous microbial enzymes, the protein was degraded constantly, resulting in the increase in MFI during storage. MFI can reflect the degradation degree of the tissue structure, and the correlation analysis indicates that it has a highly significant correlation (p<0.01) with tenderness indicator that can also reflect the degree of degradation, as a result, MFI can be used to measure the MFI beef tenderness.
(5) The solubility of the protein during storage was declining caused by the denaturation of the protein. The change of thiol group reflects the degree of oxidation of the protein; the thiol was continuously oxidized during storage to become disulfide bond, with the content decreasing. Protein solubility is significantly correlated with thiol levels (p<0.01). Fourier transform infrared spectroscopy was applied for further analysis of secondary structure content change of the soluble protein extracted, and the result showed that a-helix content of protein secondary structure decreased at the end of storage, while the content of β-pleated sheet, β-turn and random coil increased at a certain degree, was a certain, which was probably due to the protein oxidation attacked by free radical and free radical capturing the hydrogen ion, leading to breakage of hydrogen bonds, and a-helix structure was damaged and changed toward the random coil.
(6) The preservative chitosan, polyphenols and Nisin combined with CFP for storage of beef. It was found that the preservation effect of these three methods was chitosan> TP> Nisin, and there existed significant interaction between these three kinds of preservatives. Under CFP condition, the optimum compound conditions for compound preservative are: chitosan1.70%, Nisin0.03%, polyphenols0.24%. A regression model was build on Mintab15software, resulting in the regression equation of Y=12.55-1.08A-0.70B-1.08C+0.99A2+0.73B2+0.52C2+0.38AB+0.25AC+0.26BC, with model validation error of2.28%, which can better reflect the preservation effect of the compound preservative combined with CFP storage on beef. Compared with pure CFP storage, the combination of CFP storage with compound preservative combination can effectively extend the shelf life of chilled beef.
(7) The study on beef CFP storage combined with modified atmosphere packaging found that compared with the CFP storage alone, the combination of CFP storage with modified atmosphere packaging can extend the shelf life of beef for over4days, the preservation effect of50%02+50%CO2group and30%02+70%CO2group was better than that of80%02+20%CO2group. The beef TVB-N under CFP storage combined with modified atmosphere packaging was far less than the control group of CFP storage, no significant difference was found between several groups of modified atmosphere packaging. High oxygen modified atmosphere packaging will increase the rate of water loss of beef. High oxygen content gained better meat color, yet causing brown stain on the beef surface in later period of the storage, and the higher the carbon dioxide will also adversely affect the color. CFP storage combined with50%O2+50%CO2can better maintain the meat freshness.
(8) The electronic nose was used for freshness prediction of the beef stored at CFP and under normal storage. Electronic nose can well distinguish the beef of different storage times. The electronic nose builds models based on parameters of total plate count, TVB-N and sensory evaluation value, which can be used for indicator forecast of the beef not involved in modeling. The regression analysis of the predicted and measured values show the model can well predict total plate count, TVB-N and sensory evaluation value of the beef under the corresponding storage conditions. The establishment of an electronic nose model provides an effective and efficient method for determining the quality of chilled beef under different storage conditions, which is of great significance for quality control of chilled fresh beef during storage.
引文
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