CN115606630A - 一种用于减轻鲜切果蔬氧化褐变的处理方法 - Google Patents
一种用于减轻鲜切果蔬氧化褐变的处理方法 Download PDFInfo
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Abstract
本发明公开了一种减轻鲜切果蔬氧化褐变的处理方法。本发明的处理方法具体包括如下步骤:(1)果蔬处理:将新鲜果蔬消毒洗净后,鲜切后放入容器中;(2)核黄素溶液处理:将上述鲜切果蔬置于核黄素溶液中,浸泡2‑8min;(3)LED蓝光处理:将上述果蔬取出晾干后,在波长460nm蓝光下照射5‑20min,照射功率30W;(4)物料输出:将光照后的果蔬包装保存。本发明的以逆境中易发生褐变的果蔬苹果作为对象,采用核黄素介导光动力技术防褐变方法,研究表明,该方法显著抑制苹果鲜切褐变,在4℃贮藏8天仍能较好保持原有品质,减轻重量损失,有着很好的经济价值。另外,本发明方法操作简单,保鲜效果好,应用前景广。
Description
技术领域
本发明属于鲜切果蔬保鲜技术领域,具体地说,涉及一种减轻鲜切果蔬氧化褐变的处理方法。
背景技术
鲜切果蔬是一种新鲜、营养、方便、具有良好风味的食品,可满足消费者对即食健康食品的需求。但鲜切果蔬如苹果在贮藏、运输和销售过程中极易变质,这是由于果蔬经切割加工,果肉组织受损,引起各种生理生化反应。常见的有微生物病菌滋长导致营养价值降低,机械损伤导致切割面氧化褐变;呼吸速率加强加速果蔬成熟衰老,水分流失导致果实萎蔫等。这些问题都极大缩短了鲜切果蔬的货架期,为解决这些问题通常对果蔬进行提前预处理。
核黄素(Riboflavin),又名维生素B2,是异四氧嘧啶环激发产生的一种光敏剂,当氧化剂存在时,核黄素被氧化且在492~495nm或517~527nm的激发光发射范围内可以检测到荧光。核黄素具有天然、高效、安全卫生、使用剂量低、持续作用时间较长等优点被应用于医疗治疗和动物中,但在果蔬食品贮藏保鲜技术方面罕见报道,具有巨大的探索空间。
苹果果脆甘甜、营养价值高,富含矿物质和维生素,含钙量丰富,有助于代谢掉体内多余盐分,苹果酸具有可代谢热量,防止下半身肥胖等功效。其采收后呼吸代谢速度更快,酶活性更高而导致其保鲜贮藏更为困难。同时,苹果经过机械处理之后,受到严重的机械损伤,切割表面由于机体自身抗逆作用,短期内会发生褐变,严重影响产品的外观品质和经济价值。因此,在运输和贮藏期间,如何有效地控制鲜切苹果褐变并达到保鲜效果,成为苹果市场快速发展的瓶颈以及高校科研人员的研究热点。目前,国内外对减轻鲜切苹果褐变的研究报道较少。现有的减轻褐变手段主要集中在不同保鲜剂的使用上,如硫化氢、阿魏酸与芦荟凝胶或藻酸盐涂层、赖氨酸和精氨酸复配处理、姜黄素处理等,均是从酶活性抑制或底物抑制的角度分别出发,但是从膜系统稳定性以及內源抗氧化系统角度鲜有报道。也存在少量物理抑制手段的研究报道,主要有紫外线、臭氧水等,但抑制褐变效果有限。以上方法已无法满足现代果蔬加工业对高质高量、绿色环保等的加工需求。
发明内容
本发明的目的在于克服现有技术的不足,提供一种用于减轻鲜切果蔬褐变效果的处理方法,该方法安全无毒、方便卫生,能有效抑制鲜切果蔬氧化褐变,从而达到保鲜的目的。
为解决上述问题,本发明提出了一种减轻鲜切果蔬褐变的处理方法,包括如下步骤:
(1)器具用品消毒处理:将所有待使用的器具用品用0.02%NaClO溶液或75%乙醇溶液浸泡,消毒30min捞出晾干;
(2)果蔬样品处理:将预冷后的新鲜果蔬用去离子水溶液洗涤后自然晾干20min;利用切瓣工具切瓣,得到大小一致的鲜切果蔬;然后用砂布擦干鲜切果蔬表面的汁液,放入事先准备好的盘子中依次排放整齐;
(3)核黄素溶液处理:将上述鲜切果蔬置于核黄素溶液中,浸泡2-8min;
(4)光动力技术处理:将晾干后的鲜切果蔬进行蓝光LED照射处理,其中蓝光LED处理的辐照波长为460nm,照射功率为30W;照射时间为5-20min。
(5)物料输出:将光照后的果蔬,包装保存。
进一步地,上述技术方案步骤(2)中所述的鲜切果蔬优选为红富士苹果。
进一步地,上述技术方案步骤(3)中所述核黄素溶液由核黄素、蒸馏水组成。
更进一步地,上述技术方案所述的核黄素溶液采用如下方法制备而成,具体步骤如下:
事先准备好称量纸、电子天平、容量瓶、量筒、玻璃棒、药匙、烧杯、pH试纸、核黄素试剂、磁力搅拌器等,首先打开电子天平,将称量纸放在上面,用药匙从核黄素试剂中依次准确称取0.015g、0.03g和0.06g,然后分别放入烧杯中,先加入少量pH 7.0的蒸馏水用玻璃棒进行搅拌,然后缓慢加入蒸馏水,加到1L左右时,再利用磁力搅拌器进行搅拌,最后定容至2L,上下混匀,保存待用。
进一步地,上述技术方案步骤(3)中所述核黄素溶液中核黄素的浓度为20μmol L-1-80μmol L-1。
更进一步地,上述技术方案步骤(3)中所述核黄素溶液中核黄素的浓度优选为20μmol L-1。
进一步地,上述技术方案步骤(3)中所述鲜切果蔬在核黄素溶液中的浸泡时间优选为5min。
进一步地,上述技术方案步骤(5)中所述保存的温度优选为4℃。
本发明对减轻鲜切果蔬氧化褐变的机理与现有技术中减轻鲜切果蔬氧化褐变的机理不同:引起鲜切果蔬,例如苹果、梨等褐变的主要成因是多酚氧化酶(Polyphenoloxidase,缩写PPO)和过氧化物酶(Peroxidase,缩写POD)的酶促褐变,当前大多数抑制褐变方法通过外源添加PPO、POD抑制剂或者物理法抑制PPO、POD活性。因此,本发明抑制褐变有两方面因素:1.利用核黄素在LED蓝光照射后产生大量活性氧,通过诱导内源抗氧化系统,抑制PPO、POD活性;2.降低活性过氧化水平,维持膜的稳定性,抑制底物与酶接触。本发明的核黄素对鲜切苹果的褐变抑制效果十分明显,且在保鲜过程中能有效抑制PPO、POD等褐变相关酶酶活性的升高以及增强抗氧化系统如抗坏血酸(ascorbic acid,AsA)含量,抗坏血酸过氧化物酶(ascorbate peroxidase,APX)、谷胱甘肽还原酶(glutathione reductase,GR)活性,维持生物膜系统稳定性;而减轻鲜切果蔬褐变的机理主要在于两个方面的协同作用。
与现有技术相比,本发明具有如下有益效果:
(1)本发明以在逆环境中易发生褐变、贮藏期内难以保持食用品质的鲜切苹果作为研究对象,采用核黄素作为防褐变保鲜剂,研究表明,核黄素溶液对鲜切苹果具有非常显著的褐变抑制效果,而且显著抑制失重。鲜切苹果采用本发明方法处理后冷藏8天后仍能保持原有的品质和风味;
(2)本发明采用的核黄素是一种水溶性维生素,对环境无污染、安全卫生,可改善鲜切水果的食用风味,具有绿色环保、物美价廉的优点;
(3)核黄素本身作为营养价值的光敏剂,在有光源的情况下发挥作用,可高效抑制鲜切苹果褐变PPO和POD活性,同时增强抗氧化系统,维持生物膜系统稳定性,具有全面效果显著等特点。
(4)本发明不仅可以用于减轻鲜切苹果的氧化褐变,还可以应用于其他鲜切果蔬的褐变保鲜,本发明方法处理简单,保鲜效果好,创新性强,应用前景广泛。
附图说明
图1代表性L+W+、L+R20、L+R40、L+R80四种处理对鲜切苹果褐变处理效果图(其中L:蓝光LED;W:Water;R:Riboflavin,μmol);
图2L+W+、L+R20、L+R40、L+R80四种处理鲜切苹果的褐变度柱状图;
图3L+W+、L+R20、L+R40、L+R80四种处理鲜切苹果的失重率柱状图;
图4L+W+、L+R20、L+R40、L+R80四种处理鲜切苹果的多酚氧化酶(PPO)柱状图;
图5L+W+、L+R20、L+R40、L+R80四种处理鲜切苹果的过氧化氢酶(POD)柱状图;
图6L+W+、L+R20、L+R40、L+R80四种处理鲜切苹果的抗坏血酸(ASA)含量柱状图;
图7L+W+、L+R20、L+R40、L+R80四种处理鲜切苹果的抗坏血酸过氧化物酶(APX)柱状图;
图8L+W+、L+R20、L+R40、L+R80四种处理鲜切苹果的谷胱甘肽还原酶(GR)柱状图;
所有图中的数据均对应于独立重复的平均值±标准误差。(*表示P<0.05,**表示P<0.01,***表示P<0.001)。
具体实施方式
下面通过具体的实施例和附图对本发明的技术方案做进一步详细地说明。以下实施例仅是本发明较佳的实施例,并非是对本发明做其他形式的限定,任何熟悉本专业的技术人员可能利用上述揭示的技术内容加以变更为同等变化的等效实施例。凡是未脱离本发明方案内容,依据本发明的技术实质对以下实施例所做的任何简单修改或等同变化,均落在本发明的保护范围内。
本发明下述实施例中采用的核黄素溶液均由核黄素、蒸馏水组成。所述的核黄素溶液均是采用如下方法制备而成,具体步骤如下:
按比例依次准确称取核黄素,加入烧杯中,然后在磁力搅拌条件下逐滴加入蒸馏水,最终形成透明清晰、无浑浊的核黄素溶液。
以核黄素浓度为20μmol L-1的核黄素溶液的配制为例,采用如下步骤制得:
首先打开电子天平,将称量纸放在上面,用药匙从核黄素试剂中准确称取0.015g,然后分别放入烧杯中,先加入少量PH7.0的蒸馏水用玻璃棒进行搅拌,然后缓慢加入蒸馏水,加到1L左右时,再利用磁力搅拌器在转速800rpm条件下进行搅拌,最后定容至2L,上下混匀,保存待用。即为20μmol L-1浓度的核黄素溶液。
实施例1
本实施例的一种用于减轻鲜切果蔬氧化褐变的处理方法,具体包括如下步骤:
(1)器具用品消毒处理:将所有待使用的器具用品用0.02%NaClO溶液或75%乙醇溶液浸泡,消毒30min捞出晾干;
(2)苹果样品处理:将预冷后的新鲜苹果用去离子水溶液洗涤后自然晾干20min;利用切瓣工具切瓣,得到大小一致的苹果瓣;然后用砂布擦干表面的汁液,放入事先准备好的盘子中依次排放整齐;
(3)核黄素溶液处理:将上述鲜切果蔬置于20μmol L-1核黄素溶液中,浸泡5min,取出晾干;
(4)光动力技术处理:将鲜切果蔬进行蓝光LED照射处理,其中蓝光LED处理的辐照波长为460nm,照射功率为30W;照射时间为10min;
(5)物料输出:将光照后的苹果,放入保鲜盒中4℃保存。
实施例2
本实施例的一种用于减轻鲜切果蔬氧化褐变的处理方法,具体包括如下步骤:
(1)器具用品消毒处理:将所有待使用的器具用品用0.02%NaClO溶液或75%乙醇溶液浸泡,消毒30min捞出晾干;
(2)苹果样品处理:将预冷后的新鲜苹果用去离子水溶液洗涤后自然晾干20min;利用六角切瓣工具切瓣,得到大小一致的苹果瓣;然后用砂布擦干表面的汁液,放入事先准备好的盘子中依次排放整齐;
(3)核黄素溶液处理:将上述鲜切果蔬置于40μmol L-1核黄素溶液中,浸泡5min,取出晾干;
(4)光动力技术处理:将鲜切果蔬进行蓝光LED照射处理,其中蓝光LED处理的辐照波长为460nm,照射功率为30W;照射时间为10min;
(5)物料输出:将光照后的苹果,放入保鲜盒中4℃保存。
实施例3
本实施例的一种用于减轻鲜切果蔬氧化褐变的处理方法,具体包括如下步骤:
(1)器具用品消毒处理:将所有待使用的器具用品用0.02%NaClO溶液或75%乙醇溶液浸泡,消毒30min捞出晾干;
(2)苹果样品处理:将预冷后的新鲜苹果用去离子水溶液洗涤后自然晾干20min;利用切瓣工具切瓣,得到大小一致的苹果瓣;然后用砂布擦干表面的汁液,放入事先准备好的盘子中依次排放整齐;
(3)核黄素溶液处理:将上述鲜切果蔬置于80μmol L-1核黄素溶液中,浸泡5min,取出晾干;
(4)光动力技术处理:将鲜切果蔬进行蓝光LED照射处理,其中蓝光LED处理的辐照波长为460nm,照射功率为30W;照射时间为10min;
(5)物料输出:将光照后的苹果,放入保鲜盒中4℃保存。
对比例1
本对比例的一种用于减轻鲜切果蔬氧化褐变的处理方法,工艺与实施例1基本相同,区别仅在于本对照例中仅采用蒸馏水浸泡处理鲜切苹果,未用核黄素溶液处理。
实施例1-3不同方法下进行鲜切苹果贮藏过程中品质变化测试试验
苹果瓣冷藏进行一次各项指标的测定,具体如下:
褐变度测定
称取0.3g样品,加入3mL预冷的蒸馏水,于低温下匀浆2min,10000×g,4℃,离心20min,取上清液在25℃保温5min后,在波长410nm处测定上清液的吸光值A,结果以10×A表示。
失重率
贮藏过程中各试验组失重率利用标记的鲜切苹果失重率表示,采用称重法测定,并按该式计算:失重率=(m0-mi)/m0×100%式中,m0为鲜切苹果初始质量,g;mi为贮藏第i天时的鲜切苹果质量,g。
PPO活性
称取0.3g样品,置于研钵中,加入1.8mL提取缓冲液,在冰浴条件下研磨成匀浆,于4℃、12000×g离心30min,加入4.0mL50mmol L-1、pH5.5的乙酸-乙酸钠缓冲液和1mL 50mmolL-1邻苯二酚溶液,最后加入200μL酶提取液,同时立即开始计时。将反应混合液倒入比色杯中,置于分光光度计样品室中。在波长420nm处测定上清液的吸光值,作为初始值,然后每隔30s记录一次,连续测定,至少获取8个点的数据。重复三次。
POD活性
称取0.3g样品,置于研钵中,加入1.8mL提取缓冲液,在冰浴条件下研磨成匀浆,于4℃、12000×g离心30min,加入3mL 25mmol L-1愈创木酚溶液和0.5mL酶提取液,再加入0.2mL H2O2溶液混合迅速混合启动反应,同时立即开始计时。将反应混合液倒入比色杯中,置于分光光度计样品室中。在波长470nm处测定上清液的吸光值,作为初始值,然后每隔30s记录一次,连续测定,至少获取8个点的数据。重复三次。
ASA活性
称取0.2g样品置于2mL离心管中,加入20g L-1草酸溶液至2mL,摇匀,10000×g,离心10min后,取上清液备用。吸取1.5mL上清液置于100mL的三角瓶中,用已标定的2,6-二氯酚靛酚溶液滴定至出现微红色、且15s不褪色为止,记下染料的用量。同时,以1mL 20g/L草酸溶液做三次空白。
APX活性
称取0.3g样品,加入1.8mL经4℃预冷的提取缓冲液,冰浴匀浆,离心(12000×g,30min,4℃),取上清液待用。依次加入2.6mL反应缓冲液(提前放在室温)和0.1mL酶提取液。最后加入0.3mL的2mmoL/LH2O2溶液以启动酶促反应,立即混匀,立即计时。启动后15s开始记录反应体系在波长290nm处的吸光值变化,每隔15s记录一次,连续测定,至少获取8个点的数据,以蒸馏水为参比对分光光度计进行调零,重复三次。
GR活性
称取0.3g样品,加入1.8mL经4℃预冷的100mmol/L pH 7.5的磷酸缓冲液(含1mmol/L EDTA),匀浆,离心(12000×g,30min,4℃),取上清液待用。依次加入2.7mL100mmol/L pH 7.5的磷酸缓冲液(含1mmol/L EDTA)、0.1mL 5mmol/L GSSG溶液和0.2mL酶液。最后加入40μL 4mmol/L NADPH溶液以启动酶促反应,立即混匀。并开始计时,从启动后15s开始测定,记录混合液在波长340nm处的吸光值变化,每隔15s记录一次,连续测定,至少获得8个点的数据。实验重复三次。
试验统计:
采用Fisher’s least significant difference(LSD)法对不同处理样品之间进行多重比较分析(P<0.05)。
试验结果:
由图1可以看出,贮藏0-2d内,纯水光动力、20μmol L-1核黄素光动力处理组鲜切苹果瓣色泽变化尚无明显差异;贮藏第4d时,在L+W+对照组,苹果瓣已出现部分褐变,但在同期,L+R20和L+R40处理组切瓣苹果仍保持良好的白色表型;贮藏第6d时,L+W+对照和L+R80处理组苹果瓣褐变严重,而在L+R20和L+R40处理组,苹果瓣色泽较好,尤其是L+R20处理组。这些结果表明L+R20、L+R40和L+R80处理组均显著影响鲜切苹果褐变度,且最佳的控制褐变参数为L+R20即20μmol L-1核黄素光动力处理。
由图2可以看出,贮藏期间,鲜切苹果褐变度是随着贮藏时间而不断上升,表明鲜切苹果的品质是随着冷藏时间的不断延长而逐渐下降,褐变也在持续地发生。但各处理组的鲜切苹果的褐变度始终低于对照组,L+R20、L+R40处理组的效果最好,在第8d L+R20、L+R40处理组的褐变度仅为对照组的66.67%和63.15%,表明了L+R20、L+R40处理组可有效地抑制鲜切苹果产生褐变现象,从而较好的保持鲜切苹果品质。
由图3可知,鲜切苹果在贮藏过程中的失重率可影响其硬度,也能间接反映其呼吸辐照剂量,是反映鲜切果蔬品质优劣的重要指标之一。随着贮藏时间的延长,鲜切苹果失水严重,失重率逐渐增大。其中L+R20和L+R40处理组失重率均小于对照组,说明L+R20和L+R40处理组在降低失重率方面均有效。
由图4可知,贮藏期间,对照组的PPO活性呈不断增加的趋势,而处理组PPO活性呈现的是开始缓慢上升然后下降的趋势。在贮藏6d时,PPO活性达到一个最大值。其中在贮藏4d、6d、8d时,L+R20、L+R40、L+R80处理组与对照组有着明显的显著性差异(P<0.05),尤其是6d、8d。结果说明核黄素介导光动力技术能够抑制鲜切苹果PPO活性的上升。
由图5可知,贮藏期间,所有处理组的POD活性随着贮藏时间的延长呈现一直上升的趋势。贮藏8d,POD活性出现活性高峰。说明随着贮藏时间的延长,褐变发生越来越严重。自第4d起,3种处理组的POD酶活性显著(P<0.05)低于对照,但除了第2d外,3种处理组之间差异并不显著。说明贮藏过程中,3种处理组可以显著地抑制POD活性。
由图6可知,贮藏期间,苹果中的ASA含量呈现先递增后递减的趋势,其中L+R20、L+R40处理组的AsA含量显著高于对照组,表明核黄素介导光动力技术能够维持高AsA水平。
由图7可知,贮藏期间,APX酶活性呈现的是先递增后下降的趋势;在4d时其活性达到最大值。L+R20和L+R40处理组APX活性在冷藏期间均显著比对照组高,说明核黄素介导光动力技术增强了APX活性。
由图8可知,类似APX,GR酶活性呈现的是先递增后下降的趋势;在4d时其活性达到最大值。L+R20处理组GR活性在冷藏期间均显著比对照组高,说明核黄素介导光动力技术增强了GR活性。
由上述实施例及相关指标图片的抑制效果可以看出,本发明采用核黄素溶液浸泡处理对鲜切苹果的褐变进行抑制,能达到减轻褐变、抑制果蔬失重率的下降以及保鲜的目的,保持鲜切苹果的色泽品质及风味,且减轻效果是随核黄素溶液浓度升高而减弱。
Claims (6)
1.一种减轻鲜切果蔬氧化褐变的处理方法,其特征在于:具体包括如下步骤:
(1)器具用品消毒处理:将所有待使用的器具用品用0.02%NaClO溶液或75%乙醇溶液浸泡,消毒30min捞出晾干;
(2)果蔬样品处理:将预冷后的新鲜果蔬用去离子水溶液洗涤后自然晾干20min;切瓣后得到大小一致的鲜切果蔬;然后用砂布擦干鲜切果蔬表面的汁液,放入事先准备好的盘子中依次排放整齐;其中:所述的果蔬为红富士苹果;
(3)核黄素溶液处理:将上述鲜切果蔬置于核黄素溶液中,浸泡2-8min;所述核黄素溶液由核黄素、蒸馏水组成;所述核黄素溶液的浓度为20μmolL-1-80μmol L-1;
(4)光动力技术处理:将鲜切果蔬进行蓝光LED照射处理,其中蓝光LED处理的辐照波长为460nm,照射功率为30W,照射时间为5-20min;
(5)物料输出:将光照后果蔬包装保存。
2.根据权利要求1所述的用于减轻鲜切果蔬氧化褐变的处理方法,其特征在于:所述的核黄素溶液采用如下方法配制而成,具体步骤如下:
精确称取0.015g、0.03g和0.06g,然后分别放入烧杯中,先加入少量pH 7.0的蒸馏水用玻璃棒进行搅拌,然后缓慢加入蒸馏水,加到1L左右时,再利用磁力搅拌器进行搅拌,最后定容至2L,上下混匀,保存待用。
3.根据权利要求1所述的用于减轻鲜切果蔬氧化褐变的处理方法,其特征在于:步骤(3)中所述核黄素溶液的浓度为20μmol L-1。
4.根据权利要求1所述的用于减轻鲜切果蔬氧化褐变的处理方法,其特征在于:步骤(3)中所述果蔬在核黄素溶液浸泡时间为5min。
5.根据权利要求1所述的用于减轻鲜切果蔬氧化褐变的处理方法,其特征在于:步骤(4)中所述果蔬照射时间为10min。
6.根据权利要求1所述的用于减轻鲜切果蔬氧化褐变的处理方法,其特征在于:步骤(5)中所述保存的温度为4℃。
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