CN109362933A - 一种对肝脏损伤有保护作用的凉粉草多糖膏的制作方法 - Google Patents
一种对肝脏损伤有保护作用的凉粉草多糖膏的制作方法 Download PDFInfo
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Abstract
本发明涉及食品保健及食品加工领域，特别涉及一种凉粉草多糖凝胶的提取、制备及护肝作用。将新鲜的凉粉草洗净干燥、粉碎、热碱提取、离心分离、醇沉、过滤复溶、纯化得到凉粉草多糖，制备成凉粉草多糖膏，装瓶，密封。本发明产品对肝脏具有保护作用及对肝功能有调理作用，本发明产品制备的凉粉草多糖膏，不仅具有水果的清香味和蜂蜜的甜蜜感，能够满足消费者更多的口味需求，而且还具有良好的保护肝脏的功能。
Description
技术领域
本发明涉及功能性食品及食品加工领域，具体是一种对肝脏损伤有保护作用的凉粉草多糖膏，本发明产品对肝脏有保护及调理作用。
背景技术
随着我国国民经济的快速发展，人民生活水平的大幅提高，人们越来越注重于日常的生活保健，以防止疾病的发生。急性肝衰竭的治疗原则：早诊断，早治疗。在发病初期出现急性肝衰竭的典型症状，如严重的消化道症状，频繁呕吐，食欲不振，高度乏力，黄疸加深，此时进行治疗，效果较好，若当出现肝功能衰竭指标明显上升时治疗，效果甚微。因此，越早治疗对预后有重要意义。因此关于肝脏保护的食品给开发者们带来了无限商机。四氯化碳诱导小鼠产生的肝损伤，造模方法所需时间短，方法简便易行，损伤持续时间较久，是目前研究肝炎发病与治疗较为理想的模型之一。本发明采用此种造模方法来探讨凉粉草多糖对小鼠急性肝损伤的保护作用。研究结果表明，小鼠尾静脉注射四氯化碳后，引起模型组小鼠血清转氨酶的ATL和ASL显著升高，证明造模成功。凉粉草多糖的高、中剂量组与模型组比较能显著降低血清中AST和ALT的含量。尤其以凉粉草多糖高剂量组疗效更为明显，但是凉粉草多糖低剂量组疗效不显著，在试验剂量范围内显示出随着剂量的增加有疗效增强作用。
发明内容
本发明涉及一种对肝损伤有保护作用的凉粉草多糖膏，其特征在于该多糖膏包括以下步骤：
(1)将新鲜的凉粉草洗净、烘干、粉碎，取500g粉碎凉粉草用2-3L乙醇80％浸泡过夜，除去醇溶性物质，过滤，滤渣烘干，滤渣于5-10L 0.2-0.5％的碳酸钠溶液中90-95℃提取2.5-3h，过滤，滤液醇沉，过滤，将沉淀溶于超纯水中，浓缩至原始体积的三分之一，透析，浓缩，冻干，得到凉粉草多糖备用，计算出凉粉草多糖得率为凉粉草干基的7-10％；
(2)取900-1000g热水，先加入果汁40-50g和蜂蜜50-60g，再缓慢加入(1)所得的凉粉草多糖50-60g，不停的搅拌，直到冷却成膏，制得初膏；
(3)将初膏进行消毒、分装，得到凉粉草多糖膏。
所制得的凉粉草多糖膏对肝脏损伤有保护作用。
所述步骤(1)中，通过加入无水乙醇将溶液调节至最终浓度为80％(v/v)并在25℃下保持过夜。
所述步骤(1)中，所述果汁选取芒果汁、苹果汁、柠檬汁。
本发明的有益效果：
(1)步骤(1)中，碳酸钠提取，可以提高凉粉草多糖的得率。
(2)本发明所制备的凉粉草多糖膏对肝脏损伤有保护作用。
具体实施方式
实施例1
(1)将新鲜的凉粉草洗净、烘干、粉碎，取500g粉碎凉粉草用2L乙醇(80％)浸泡过夜，除去某些醇溶性物质，过滤，滤渣烘干，滤渣于5L 0.2％的碳酸钠溶液中90℃提取3h，过滤，滤液醇沉，过滤，将沉淀溶于超纯水中，浓缩至原始体积的三分之一，透析，浓缩，冻干，得到凉粉草多糖备用。计算出凉粉草多糖得率为凉粉草干基的7％；
(2)将(1)中的凉粉草多糖粉末溶解于超纯水中，得到凉粉草多糖溶液用于肝脏损伤保护作用试验，凉粉草多糖对肝脏损伤具有保护作用；(3)取1000g热水，先加入果汁50g和蜂蜜60g，再缓慢加入(1)所得的凉粉草多糖50g，不停的搅拌，直到冷却成膏，制得初膏；
(4)将初膏消毒、分装，得到凉粉草多糖膏。
实施例2
(1)将新鲜的凉粉草洗净、烘干、粉碎，取500g粉碎凉粉草用3L乙醇(80％)浸泡过夜，除去某些醇溶性物质，过滤，滤渣烘干，滤渣于10L 0.2％的碳酸钠溶液中95℃提取2.5h，过滤，滤液醇沉，过滤，将沉淀溶于超纯水中，浓缩至原始体积的三分之一，透析，浓缩，冻干，得到凉粉草多糖备用。计算出凉粉草多糖得率为凉粉草干基的10％；
(2)将(1)中的凉粉草多糖粉末溶解于超纯水中，得到凉粉草多糖溶液用于肝脏损伤保护作用试验，凉粉草多糖对肝脏损伤具有保护作用；
(3)取900g热水，先加入果汁40g和蜂蜜50g，再缓慢加入(1)所得的凉粉草多糖60g，不停的搅拌，直到冷却成膏，制得初膏；
(4)将初膏消毒、分装，得到凉粉草多糖膏。
对肝脏损伤治疗的实验：
1.材料与仪器
1.1受试物与药品
凉粉草多糖：将新鲜的凉粉草洗净干燥、粉碎、热碱提取、离心分离、醇沉、过滤复溶、纯化得到凉粉草多糖备用，本产品肝脏有保护作用及对肝功能有调理作用。四氯化碳(西陇科学股份有限公司)；联苯双酯滴丸(浙江医药股份有限公司)；ALT试剂盒(南京建成生物工程研究所)；AST试剂盒(南京建成生物工程研究所)。
1.2动物
清洁级昆明种雄性小鼠，湖南斯莱克公司提供，体重18-25g。环境温度：25±2℃，12h光照周期的动物室内饲养，自由饮水、进食。实验前饲养1周以适应环境。
1.3仪器
冷冻离心机(Sigma公司)；多功能全自动酶标仪(美国Thermo公司)；ALPHA 1-4冷冻干燥机(德国CHRIST公司)；FA1104电子分析天平(上海梅特勒-托利多仪器公司)；HH4-数显恒温水浴锅(常州华普达仪器有限公司)。
2.方法
2.1动物模型的建立
采用四氯化碳诱导小鼠急性肝损伤。小鼠尾静脉注射0.2％四氯化碳0.2mL/20g以诱导肝损伤，正常组小鼠尾静脉注射同等剂量生理盐水。
2.2动物分组与处理
将实验动物随机分为7组：正常组、模型组、凉粉草多糖高(300mg/kg)、中(200mg/kg)、低(100mg/kg)剂量组、联苯双酯滴丸组(200mg/kg)，凉粉草多糖+联苯双酯滴丸(100mg/kg)，每组10只。小鼠适应7天后，连续灌胃28天，最后一天将所有组别小鼠空腹过夜后，除正常组小鼠外，所有小鼠腹腔注射0.2％四氯化碳花生油溶液0.2mL/20g，正常组给予同等剂量生理盐水。造模结束后小鼠禁食12h左右，称重后眼球采血，脱椎处死取肝脏和血液，血液3000转10min离心，取血清超低温冰箱冻存以备下一步检测所需。
2.3血清中ALT、AST含量检测
取小鼠血清按试剂盒说明检测。
2.4统计学分析
采用SPSS21.0统计分析软件进行t检验，数据用“均数±标准差”表示，双侧检验以p<0.05表示差别有统计学意义。
3.结果
3.1对小鼠免疫性肝损伤的治疗保护作用
与正常对照组比，模型组血清AST、ALT水平显著升高(p<0.05)，表明四氯化碳干预后对小鼠肝细胞造成了严重的损伤，动物模型建立成功。凉粉草多糖高、中、低剂量组与模型组相比有显著差异(p<0.05)。阳性药物对照组与模型组相比有显著差异(p<0.05)。结果见表1。
表1凉粉草多糖对急性肝损伤小鼠血清转氨酶的影响(n＝12)
4.讨论
本次试验表明凉粉草多糖膏对急性肝损伤有保护作用及对肝功能有调理作用。
表1凉粉草多糖膏对免疫性肝损伤小鼠血清转氨酶的影响(n＝12)
该值表示为三次测定的平均值±标准偏差。不同字母a-e表示有显著差异(p<0.05)。从上述数据可以看出本发明所制备的凉粉草多糖膏对肝脏的损伤有明显的治愈效果。
Claims (4)
1.一种对肝脏损伤有保护作用的凉粉草多糖膏的制作方法，其特征在于，包括以下步骤：
(1)将新鲜的凉粉草洗净、烘干、粉碎，取500g粉碎凉粉草用2-3L乙醇80％浸泡过夜，除去醇溶性物质，过滤，滤渣烘干，滤渣于5-10L 0.2-0.5％的碳酸钠溶液中90-95℃提取2.5-3h，过滤，滤液醇沉，过滤，将沉淀溶于超纯水中，浓缩至原始体积的三分之一，透析，浓缩，冻干，得到凉粉草多糖备用，计算出凉粉草多糖得率为凉粉草干基的7-10％；
(2)取900-1000g热水，先加入果汁40-50g和蜂蜜50-60g，再缓慢加入(1)所得的凉粉草多糖50-60g，不停的搅拌，直到冷却成膏，制得初膏；
(3)将初膏进行消毒、分装，得到凉粉草多糖膏。
2.根据权利要求1所述的对肝脏损伤有保护作用的凉粉草多糖膏的制备方法，其特征在于：所制得的凉粉草多糖膏对肝脏损伤有保护作用。
3.根据权利要求1所述的对肝脏损伤有保护作用的凉粉草多糖膏的制备方法，其特征在于：步骤(1)中，通过加入无水乙醇将溶液调节至最终浓度为80％(v/v)并在25℃下保持过夜。
4.根据权利要求1所述的对肝脏损伤有保护作用的凉粉草多糖膏的制备方法，其特征在于：步骤(1)中，所述果汁选取芒果汁、苹果汁、柠檬汁。
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