壳聚糖-鱼精蛋白复合涂膜的制备及保鲜效果研究
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摘要
试验探究壳聚糖-鱼精蛋白复合涂膜对罗非鱼片冷藏保鲜的切实性。首先考察醋酸浓度对壳聚糖抑菌效果的影响,然后测定壳聚糖和鱼精蛋白溶液单独作用的最小抑菌浓度(MIC),通过药物复配得到的分级抑菌指数(FIC)确定复合制剂配比,最后研究涂膜内药物释缓效果及抗菌性能来探讨复合涂膜在罗非鱼片冷藏保鲜的应用。试验结果表明,壳聚糖和鱼精蛋白单独作用时的MIC对大肠杆菌为0.625 0和1.250 0 mg·mL~(-1),对金黄色葡萄球菌为0.312 5和0.625 0 mg·mL~(-1)。FIC指数确定药剂最优剂量比为1∶4;壳聚糖-鱼精蛋白复合涂膜对罗非鱼片有明显的冷藏保鲜作用,涂膜内复合药物能在细菌快速增长期前基本释放,可有效控制罗非鱼片TVB-N值、pH的上升和鱼片表面细菌总数的增长。复合药剂涂膜保鲜效果明显优于对照组,于4℃±1℃贮藏10 d后其各项指标均处于可接受范围内,较对照组延长货架期6 d左右。
This study focused on the feasibility of chitosan-protamine composite in Tilapia fillet preservation. First, find the concentration of the acetic acid to dissolve chitosan by bacteriostatic ring test; Then, investigate the minimal inhibitory concentration(MIC) of chitosan and protamine alone by broth microdilution method, and on this basis determining the proportion of the complex through combination of the drugs; Finally, focus on the containing drugs release effect in membrane, and study the application of composite coating to the fresh-keeping Tilapia fillets by the test of antibacterial properties. The results showed that when chitosan and protamine acted alone, the MIC value was 0.625 0 and 1.250 0 mg·mL~(-1) for escherichia coli, and the MIC value was 0.312 5 and 0.625 0 mg·mL~(-1) for Staphylococcus aureus. Chitosan and protamine's optimum proportion was determined to be 1︰4 by fractional inhibitory concentration test. Chitosan-protamine composite coating obviously had a fresh-keeping refrigeration effect on Tilapia fillets. The compound drugs in the coating could basically release before the rapid growth period of bacteria, which could effectively control the increase of the TVB-N value, the pH and the total number of the bacteria on the surface. The preservation effect of the medicament mixture group was better than the matched group. Under the 4 ℃±1 ℃ storage condition, the fillets could be preserved for 10 d with each measurement index which was within the acceptable range. Thus the shelf life of tilapia fillets could be extended for 6 d.
This study focused on the feasibility of chitosan-protamine composite in Tilapia fillet preservation. First, find the concentration of the acetic acid to dissolve chitosan by bacteriostatic ring test; Then, investigate the minimal inhibitory concentration(MIC) of chitosan and protamine alone by broth microdilution method, and on this basis determining the proportion of the complex through combination of the drugs; Finally, focus on the containing drugs release effect in membrane, and study the application of composite coating to the fresh-keeping Tilapia fillets by the test of antibacterial properties. The results showed that when chitosan and protamine acted alone, the MIC value was 0.625 0 and 1.250 0 mg·mL~(-1) for escherichia coli, and the MIC value was 0.312 5 and 0.625 0 mg·mL~(-1) for Staphylococcus aureus. Chitosan and protamine's optimum proportion was determined to be 1︰4 by fractional inhibitory concentration test. Chitosan-protamine composite coating obviously had a fresh-keeping refrigeration effect on Tilapia fillets. The compound drugs in the coating could basically release before the rapid growth period of bacteria, which could effectively control the increase of the TVB-N value, the pH and the total number of the bacteria on the surface. The preservation effect of the medicament mixture group was better than the matched group. Under the 4 ℃±1 ℃ storage condition, the fillets could be preserved for 10 d with each measurement index which was within the acceptable range. Thus the shelf life of tilapia fillets could be extended for 6 d.
引文
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[11]张雪娇,齐暑华,刘春叶,等.茚三酮显色法测定壳聚糖含量[J].化学研究与应用,2015,27(3):407-411.
[12]张凤雪,文娥,李柯翱,等.维药制剂药渣中蛋白质及游离氨基酸的含量测定[J].世界中医药,2016,11(2):327-329.
[13]韩婷,史国华,李素哲,等.壳聚糖中蛋白质含量测定方法的研究[J].中国医疗器械杂志,2016,40(2):122-124.
[14]郑连英,朱江峰,孙昆山.壳聚糖的抗菌性能研究[J].材料科学与工程,2000(2):22-24.
[15]吴迪,刘辉,浦金辉,等.O-羧甲基壳聚糖抑菌作用研究[J].医药导报,2013,32(2):142-145.
[16]谢勇,周南进,李弼民,等.不同浓度和pH的壳聚糖对幽门螺杆菌抑菌作用的影响[J].中华微生物学和免疫学杂志,2002(1):17.
[17]ANON.New food preservative[J].Journal of Seafood International,1987,2(10):48.
[18]孙慎侠,王海凤,齐珊珊.壳聚糖及其单体体外抑菌作用研究[J].大连大学学报,2004(2):97-98,109.
[19]ROLER S,COVILL N.The antifungal properties of chitosan in laboratory media and apple juice[J].International Journal Food Microbiol,1999(47):67-77.
[20]毕红燕,陈峥宏,綦廷娜,等.质子泵抑制剂与抗生素对幽门螺杆菌的联合抗菌作用[J].世界华人消化杂志,2014,22(23):3547-3551.
[21]史同瑞,刘宇,苏永福,等.壳聚糖-海藻酸钠微囊形成机理及制备方法研究进展[J].上海畜牧兽医通讯,2015(5):11-13.
[2]匡银近,张利芳,覃彩芹,等.可食性壳聚糖复合涂膜包装对鲜切莲藕保鲜效果的影响[J].湖北工程学院学报,2014,34(6):25-28.
[3]HONARKAR H,BARIKANI M.Applications of biopolymers I:Chitosan[J].Monatsh Chem,2009(7):1403-1420.
[4]李文茹,谢小保,欧阳友生,等.鱼精蛋白抑菌机理及在食品防腐中的应用[J].微生物学通报,2007(4):795-798.
[5]POTTER R,TRUELSTRUP HANSEN L,GILL T A.Effect of electrostatic interactions on the inhibition of foodborne bacteria by modified and native protamine[J].International Journal of Food Microbiology,2005(103):23-34.
[6]王南舟,钟立人,黄高雄,等.鱼精蛋白抗菌特性的研究[J].食品科学,2000(4):43-46.
[7]廖晓珊,钟晓红.壳聚糖协同竹醋液的抗菌性研究[J].中国食品添加剂,2009(6):126-130.
[8]郭锦棠,张瑾,殷俊威.抗菌性海藻酸钠膜的制备及性能分析[J].天津大学学报(自然科学与工程技术版),2013,46(7):653-658.
[9]邓靖,谭兴和,周晓媛.丁香油-海藻酸钠可食性抗菌膜的研制[J].食品工业科技,2009,30(6):302-305.
[10]AMIDON G L,LENNERN H,SHAH V P,et al.Atheoretical basis for a biopharmaceutic drug classification:The correlation of in vitro drug product dissolution and in vivo bioavailability[J].Pharmaceutical Research,1995(3):413-420.
[11]张雪娇,齐暑华,刘春叶,等.茚三酮显色法测定壳聚糖含量[J].化学研究与应用,2015,27(3):407-411.
[12]张凤雪,文娥,李柯翱,等.维药制剂药渣中蛋白质及游离氨基酸的含量测定[J].世界中医药,2016,11(2):327-329.
[13]韩婷,史国华,李素哲,等.壳聚糖中蛋白质含量测定方法的研究[J].中国医疗器械杂志,2016,40(2):122-124.
[14]郑连英,朱江峰,孙昆山.壳聚糖的抗菌性能研究[J].材料科学与工程,2000(2):22-24.
[15]吴迪,刘辉,浦金辉,等.O-羧甲基壳聚糖抑菌作用研究[J].医药导报,2013,32(2):142-145.
[16]谢勇,周南进,李弼民,等.不同浓度和pH的壳聚糖对幽门螺杆菌抑菌作用的影响[J].中华微生物学和免疫学杂志,2002(1):17.
[17]ANON.New food preservative[J].Journal of Seafood International,1987,2(10):48.
[18]孙慎侠,王海凤,齐珊珊.壳聚糖及其单体体外抑菌作用研究[J].大连大学学报,2004(2):97-98,109.
[19]ROLER S,COVILL N.The antifungal properties of chitosan in laboratory media and apple juice[J].International Journal Food Microbiol,1999(47):67-77.
[20]毕红燕,陈峥宏,綦廷娜,等.质子泵抑制剂与抗生素对幽门螺杆菌的联合抗菌作用[J].世界华人消化杂志,2014,22(23):3547-3551.
[21]史同瑞,刘宇,苏永福,等.壳聚糖-海藻酸钠微囊形成机理及制备方法研究进展[J].上海畜牧兽医通讯,2015(5):11-13.