CN117256772B - 林蛙油的保鲜方法 - Google Patents
林蛙油的保鲜方法 Download PDFInfo
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Abstract
本申请提供了一种林蛙油的保鲜方法，其包括：使用以鲜梨、生姜、绿茶和高度白酒制备的脱腥液对林蛙油鲜体脱腥处理；低温脱水处理；喷洒含抗菌肽的保鲜剂进行防腐处理。本申请的林蛙油保鲜方法基本使用天然材料进行，绿色安全，制备的微干品林蛙油保存性能好，食用方便；且能保留大比例的营养成分。
Description
技术领域
本申请属于食品保鲜和生物技术领域，具体地，本申请提供了一种林蛙油的保鲜方法。
背景技术
中国林蛙（Rana temporaria chensinensis, David）主要分布于中国东北地区和朝鲜。雌性中国林蛙的输卵管部分被称为林蛙油，是传统名贵药材，其有补肾益精、养阴润肺、抗氧化、提高免疫力、增强性功能等功效。目前已经采用现代医学、生物学手段对林蛙/林蛙油的功效进行了大量探索和验证。
林蛙油的保鲜一直是业界的难题，林蛙油在保存中很容易出现染菌腐败、氧化酸败等问题。为满足长期保存的要求，林蛙油一般被制成干品出售，干品林蛙油食用时需要经过数小时到数十小时的长时间泡发过程，过程繁琐，不能很好地适应现代高节奏生活的需求；而使用大量使用防腐剂、抗氧化剂等制备即食林蛙油不仅会破坏产品的绿色健康概念，而且对产品的风味常有不良影响。
因此，本领域中有开发有效抑菌、抗酸败并且食用相对方便的林蛙油产品的需求。
发明内容
一方面，本申请提供了一种林蛙油的保鲜方法，所述保鲜方法包括：
（1）使用以鲜梨、生姜、绿茶和高度白酒制备的脱腥液对林蛙油鲜体脱腥处理得到脱腥处理的林蛙油；
（2）对（1）得到的脱腥处理的林蛙油进行低温脱水处理得微干品林蛙油；
（3）向（2）得到的微干品林蛙油喷洒保鲜剂进行防腐处理；
进一步地，步骤（2）包括将脱腥处理的林蛙油置于烘干机内，20-25℃低温脱水20-30分钟。
进一步地，所述脱腥液按照以下方法制备：
将鲜梨2-5重量份、生姜2-5重量份、绿茶1-4重量份于破壁机中粉碎成浓浆；向浓浆中加入酶和乳化剂，酶处理一定时间；加入60-100重量份60-65度白酒中浸提8-24小时，取上清液作为脱腥液。
进一步地，所述酶为纤维素酶或木聚糖酶；所述乳化剂为氢化松香甘油酯或乳酸脂肪酸甘油酯。
进一步地，向浓浆中加入200-400 U/mL浓浆的木聚糖酶，以及10-20mg/mL浓浆的氢化松香甘油酯或乳酸脂肪酸甘油酯，酶处理60-12分钟。
进一步地，所述脱腥液按照以下方法制备：
将鲜梨4重量份、生姜4重量份、绿茶3重量份于破壁机中粉碎成浓浆；向浓浆中加入300 U/mL浓浆的木聚糖酶以及20mg/mL浓浆的乳酸脂肪酸甘油酯，混合均匀后酶处理90分钟；加入80重量份60度白酒中浸提12小时，取上清液作为脱腥液。
基于所用酶的不同活性，特别是在酒精中的活性，酶处理后可以直接加入白酒或者增加高温灭酶的步骤，高温灭酶的条件可以参考本领域相关书籍。
进一步地，所述保鲜剂包含山梨酸钾、表面活性剂、吐温80以及抗菌肽。
所述含抗菌肽优选各种动物植物材料来源的，通过酶解、纯化、浓缩等方式制备的抗菌肽，其可以为单一肽的纯品、也可以为多种肽和其他成分的混合物；其可以为固体形式、也可以为提取液或其浓缩液的液体形式。
进一步地，所述抗菌肽为林蛙抗菌肽的浓缩液，其按照以下方法制备：（1）取脱脂除卵的林蛙干体50g；（2）向林蛙干体中加5%醋酸500mL，于10℃溶胀15分钟，得到溶胀后的林蛙干体和5%醋酸的混合物；（3）将（2）得到的溶胀后的林蛙干体和5%醋酸的混合物，放入功率为100W，频率为20kHz的超声波中提取15分钟，过滤得提取液和固体；（4）向步骤（3）所得固体中加5%醋酸500mL，放入功率为100W，频率为20kHz的超声波中提取15分钟，过滤得提取液；（5）合并（3）和（4）所得提取液，在3000rpm离心20分钟，弃去沉淀，收集上清；（6）将上清在50℃环境下加热5min后降至10℃，过滤得滤液；（6）将（5）所得滤液35℃，-0.5个大气压条件下浓缩至1/3体积得到林蛙抗菌肽浓缩液。
进一步地，所述保鲜剂为包含0.6%w/v山梨酸钾，1%w/v硬脂酸钾、0.8%w/v吐温80、40%v/v林蛙抗菌肽浓缩液的水溶液。
本申请所述的林蛙油鲜体是指从活体林蛙上经过宰杀解剖或活体取样获得的雌性林蛙输卵管部分，除必要的去除筋膜等杂质的步骤，不经过任何其他物理、化学、生物加工。
具体实施方式
实施例1基础处理和制剂配制
脱腥液：
将鲜梨4重量份、生姜4重量份、绿茶3重量份于破壁机中粉碎成浓浆，加入80重量份60度白酒中浸提12小时，取上清液作为脱腥液。
防腐保鲜剂：
参照本申请申请人早期授权专利ZL201310044262.3说明书实施例1的方法制备林蛙抗菌肽：（1）取脱脂除卵林蛙干体50g，进行去尘除杂处理；（2）向林蛙干体中加5%醋酸500mL于10℃溶胀15分钟；（3）将溶胀后的蛙体和5%醋酸液，一起放入功率为100W，频率为20kHz的超声波中提取15分钟，过滤得提取液；（4）将过滤所得固体中加5%醋酸500mL，放入功率为100W，频率为20kHz的超声波中提取15分钟，过滤得提取液；（5）合并提取液，在3000rpm离心20分钟，弃去沉淀，收集上清；（6）将上清在50℃环境下加热5min后降至10℃，过滤；（6）35℃，-0.5个大气压条件下浓缩至1/3体积得到林蛙抗菌肽浓缩液。
配制含0.6%w/v山梨酸钾，1%w/v硬脂酸钾、0.8%w/v吐温80、40%v/v林蛙抗菌肽浓缩液的防腐保鲜剂溶液。
基本制备步骤
取林蛙油鲜体：
秋季于吉林基地回捕中国林蛙（Rana temporaria chensinensis, David），解剖处理得到林蛙油，去除黑色筋膜等杂质后作为林蛙油鲜体备用。
脱腥处理：
将林蛙油鲜体浸泡于脱腥液中8小时，取出后淋干表面水分。
脱水处理：
将脱腥处理的林蛙油置于烘干托盘上，烘干托盘置于烘干机内，25℃低温脱水30分钟，至林蛙油表面微干，表面形成干燥膜体，用指压触感软弹，无表面渗出水份。
防腐处理：
在脱水处理获得的微干林蛙油上均匀喷洒防腐保鲜剂（约200-400g微干品林蛙/100mL防腐保鲜剂），自然风干，称量分装真空塑封。食用真空塑封林蛙油取出，清水洗净表面，微泡发后蒸食或煮食时即可。
实施例2基本检测方法
酸价和过氧化值的测定：
将林蛙油吸去表面水分，用滤纸包裹；加入索氏抽提器中，加入石油醚；在60摄氏度水浴中提取4小时；回收溶剂。基本按照GB/T 5530-2005（动植物油脂 酸值和酸度测定）中的热乙醇法测定酸价；基本按照GB/T 5009.37-2003（食用植物油卫生标准分析方法）中的比色法测定过氧化值（均为3份取平均）。
抗氧化能力测定：
将脱腥液离心取上清，取脱腥液样品2.5mL，与2.5mL 0.1mM的DPPH无水乙醇溶液（样品）或者2.5mL无水乙醇混合（对照），25℃下避光反应30min；以分光光度计测定样品反应液、对照反应液以及参比反应液（2.5mL无水乙醇加2.5mL 0.1mM的DPPH无水乙醇溶液）517nm处的吸光度（3份取平均）。
DPPH自由基清除率%=（1-（A样品-A对照）/A参比）。
实施例3脱腥液制备中的酶处理
申请人使用鲜梨、生姜、绿茶和白酒制备脱腥液，并以之处理林蛙油鲜体来制备微干品林蛙油。结果表明，该方法制备的产品口感良好；除快速有效脱除过多水份外，可减少林蛙油水溶性蛋白质和脂肪酸流失，微干品林蛙油的水溶性蛋白质含量可达到38.92%，而未加工林蛙油水溶性蛋白质含量在39.53%（参考NY/T1205-2006，水提水溶性蛋白后凯氏定氮法测定）；而且林蛙抗菌肽的保鲜剂可以有效抑制产品腐败，可以满足保存所需。但该产品在长期保存后容易产生哈油味，口感上恶化明显，不及一般干品。
为解决此问题，申请人尝试使用酶解方式提高脱腥液的抗氧化效果，进而抑制酸败。尝试了大量植物/中药酶解可用的酶及其用量（实验用酶均为食品级，来自），典型的酶带来的自由基清除效果如表1所示（酶用量以U/mL浓浆计，浓浆pH值未做调整直接加入酶，酶处理时间为90分钟，加入大量高度白酒后酶可以认为基本失活；仅展示效果较好/经济的优选浓度结果，每种酶的浓度筛选实验未展示）。
表1酶处理后脱腥液的抗氧化性能
结果表明，木聚糖酶和纤维素酶均能实现自由基清除率的有效提升，但按照一般植物提取物DPPH自由基清除效果的经验，50%的自由基清除率并不理想，如果要达到抑制酸败的效果，需要进一步改进酶解体系。
随后，申请人尝试在酶解体系中加入乳化剂以改善酶解效果，尝试了木聚糖酶和纤维素酶与多种乳化剂的配合（乳化剂用量实验结果未展示，仅展示最佳用量下的结果）。除了检测DPPH自由基清除率之外，部分技术方案还直接制备产品进行了保存实验（不加包装，40摄氏度，60%湿度，保存21日后检测酸价和过氧化值）。
表2 纤维素酶组结果
注：新制备的产品直接测定酸价为2.0mg/g，过氧化值在0.19g/100g的水平。
表3 木聚糖酶组结果
注：新制备的产品直接测定酸价为2.0mg/g，过氧化值在0.19g/100g的水平。
结果表明，加入一定量的氢化松香甘油酯或乳酸脂肪酸甘油酯可以有效提高脱腥液酶处理的效果，虽然木聚糖酶组自由基清除率整体上不及纤维素酶，但其实际保鲜效果上却优于纤维素酶组，这表明酸价和过氧化值的变化并不能简单地以自由基清除率的变化（抗氧化性）来预期，酶解出的多糖结构/多糖存在方式等因素也可能影响酸价和过氧化值的变化。
Claims (4)
1.一种林蛙油的保鲜方法，其特征在于，所述保鲜方法包括：
（1）使用以鲜梨、生姜、绿茶和高度白酒制备的脱腥液对林蛙油鲜体脱腥处理得到脱腥处理的林蛙油；
（2）对步骤（1）得到的脱腥处理的林蛙油进行低温脱水处理得微干品林蛙油；
（3）向步骤（2）得到的微干品林蛙油喷洒保鲜剂进行防腐处理；
其中，步骤（1）中所述脱腥液按照以下方法制备：
将鲜梨2-5重量份、生姜2-5重量份、绿茶1-4重量份于破壁机中粉碎成浓浆；向浓浆中加入200-400 U/mL浓浆的木聚糖酶，以及10-20mg/mL浓浆的氢化松香甘油酯或乳酸脂肪酸甘油酯，酶处理60-12分钟；加入60-100重量份60-65度白酒中浸提8-24小时，取上清液作为脱腥液；
所述保鲜剂为包含0.6%w/v山梨酸钾，1%w/v硬脂酸钾、0.8%w/v吐温80、40%v/v林蛙抗菌肽浓缩液的水溶液。
2.根据权利要求1所述的保鲜方法，其中步骤（2）包括将脱腥处理的林蛙油置于烘干机内，20-25℃低温脱水20-30分钟。
3.根据权利要求1所述的保鲜方法，其中步骤（1）中所述脱腥液按照以下方法制备：
将鲜梨4重量份、生姜4重量份、绿茶3重量份于破壁机中粉碎成浓浆；向浓浆中加入300U/mL浓浆的木聚糖酶以及20mg/mL浓浆的乳酸脂肪酸甘油酯，混合均匀后酶处理90分钟；加入80重量份60度白酒中浸提12小时，取上清液作为脱腥液。
4.根据权利要求1-3任一项所述的保鲜方法，其中抗菌肽为林蛙抗菌肽的浓缩液，其按照以下方法制备：（1）取脱脂除卵的林蛙干体50g；（2）向林蛙干体中加5%醋酸500mL，于10℃溶胀15分钟，得到溶胀后的林蛙干体和5%醋酸的混合物；（3）将（2）得到的溶胀后的林蛙干体和5%醋酸的混合物，放入功率为100W，频率为20kHz的超声波中提取15分钟，过滤得提取液和固体；（4）向步骤（3）所得固体中加5%醋酸500mL，放入功率为100W，频率为20kHz的超声波中提取15分钟，过滤得提取液；（5）合并（3）和（4）所得提取液，在3000rpm离心20分钟，弃去沉淀，收集上清；（6）将上清在50℃环境下加热5min后降至10℃，过滤得滤液；（6）将（5）所得滤液35℃，-0.5个大气压条件下浓缩至1/3体积得到林蛙抗菌肽浓缩液。
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