CN114158621A - 一种枸杞果油和核桃油的组合物及其应用 - Google Patents
一种枸杞果油和核桃油的组合物及其应用 Download PDFInfo
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Abstract
本发明涉及功能食品制备技术领域,具体涉及一种枸杞果油和核桃油的组合物及其应用;组合物中包含有任一比例的枸杞果油和核桃油,组合物中还添加有具有抗疲劳效果的蛋白肽;本发明将枸杞果油和核桃油配合使用,制备的组合物具有缓解视力疲劳的功能。同时将金枪鱼肉使用Alcalase2.4酶和胰蛋白酶的复合酶进行酶解,制备出具有抗疲劳功能的蛋白肽,将所制备的蛋白肽添加到枸杞果油和核桃油制成的溶剂,制成的制品即具有抗视力疲劳的功效,也具有缓解疲劳的效果。
Description
技术领域
本发明属于功能食品制备技术领域,具体涉及一种枸杞果油和核桃油的组合物及其应用。
背景技术
视力疲劳是指从事近距离工作或学习,由于过度使用眼睛而产生的眼睛疲劳。此症常发于从事近距离精密工作、电脑工作,或者在照明条件不佳(光线过亮/过暗)的环境内学习或工作,以及患有近视、远视、老光等屈光不正及身体衰弱的人。视力疲劳的症状有眼疲劳、眼干涩、异物感、眼皮沉重感、视物模糊、畏光流泪、眼胀痛及眼部充血等,严重者还可出现头痛、头昏、恶心、精神萎靡、注意力不集中、记忆力下降、以及食欲不振等全身症候群,少数患者可出现复视、立体视觉功能障碍、眼压升高、角膜损害等,青少年还可以出现近视或加深近视程度。有青光眼、眼表面或眼前节疾患者还可因眼的过度疲劳而引发或加重原有眼病,因而缓解视力疲劳一直是研究热点。
目前,不少学者在缓解视力疲劳方面做了大量工作,但是各种缓解视力疲劳的措施在效果上各有不尽如人意的地方,开发的具有抗视力疲劳作用的中药、化药等,长期服用存在一定的副作用。
发明内容
本发明的目的在于提供一种枸杞果油和核桃油的组合物及其应用,所使用枸杞果油和核桃油具有缓解视力疲劳的作用;而添加的蛋白肽具有抗疲劳的作用,从而弥补现有技术的不足。
本发明首先提供一种枸杞果油和核桃油的组合物,所述的组合物中包含有任一比例的枸杞果油和核桃油;
更进一步的,所述组合物中还添加有具有抗疲劳效果的蛋白肽;
所述的具有抗疲劳效果的蛋白肽,作为实施例的一种具体记载,是金枪鱼肉的Alcalase2.4酶和胰蛋白酶的酶解液进行离心获得的;
所述的酶解液,其酶解温度为35℃的条件下酶解5-6h;
其中Alcalase2.4酶和胰蛋白酶的酶活比为1:3;
更进一步的,所述的蛋白肽的分子量为500-3000Da。
所述的蛋白肽,其中一种多肽的氨基酸序列为YYMEGDFAFGDNY。
更进一步的,所述的组合物中还包含常用的调味料,作为优选,为甜味剂或酸味剂;
本发明还提供了上述技术方案所述制备方法制备得抗疲劳制品。
本发明将枸杞果油和核桃油配合使用,制备的组合物具有缓解视力疲劳的功能。同时将金枪鱼肉使用Alcalase2.4酶和胰蛋白酶的复合酶进行酶解,制备出具有抗疲劳功能的蛋白肽,将所制备的蛋白肽添加到枸杞果油和核桃油制成的溶剂,制成的制品即具有抗视力疲劳的功效,也具有缓解疲劳的效果。
附图说明
图1:50%乙醇洗脱组分的HPLC分离图;
图2:30-32min洗脱液的纯化检测图;
图3:不同剂量PA1多肽对小鼠负重游泳时间的影响图。
具体实施方式
本发明将具有抗疲劳功能的蛋白肽添加到枸杞果油和核桃油制成的溶剂中,使制成的可食用的制品即具有抗视力疲劳的功效,也具有缓解疲劳的效果。
下面结合实施例和附图对本发明进行详细的描述
实施例1:制备枸杞果油的方法
本实施例的枸杞果油的制备方法,包括如下的步骤:
(1)挑选无破碎粒、果实饱满的优质枸杞子,置于低温粉碎机中粉碎并通过40目筛,得到的枸杞粉末称取5kg,装入提取布袋并系好口,置入亚临界萃取设备的密闭萃取罐中,关闭进料通道,抽真空至-0.075MPa,泵入20L的丁烷,加热至35℃,设置萃取压力0.5MPa、超声频率为15KHz下进行萃取,萃取时间25min,萃取3次;
(2)萃取结束后,将萃取溶剂抽出至分离罐中,开启真空泵,将分离罐的压力降至-0.06MPa,温度保持在35℃,蒸发物料中的丁烷,并将气化的丁烷经隔膜压缩机压缩成液态,回流至溶剂罐中,关闭真空泵,使分离罐的压力达到0MPa,从净化口排出枸杞果油,制得枸杞果油。
本发明所使用的枸杞果油可以采用市售的任一款枸杞果油产品。
实施例2制备核桃油的方法
(1)取鲜的油核桃果清洗、粉碎后至20目,加入40℃重量为鲜果4倍重量的纯净水,搅拌;
(2)用高速剪切对步骤(1)中果肉中细胞组织进一步破碎提取,高速剪切转速为12000r/min,提取时间为8min,每提取2min,间隔0.5min;
(3)高速剪切破碎后的果肉水溶液用搅拌器融合形成果浆,在35℃温度下加热保温搅拌;
(4)然后在常温下压榨处理步骤(3)中的果浆;
(5)用高速离心机(高速离心机的转速为3000r/min,时间为25min)对压榨后果浆进行固液分离,分离果渣后,对油水混合液进一步用200目的滤膜过滤;
(6)在油水混合液中加入油水混合物重量2%的氯化钠,搅拌使得氯化钠完全溶解;
(7)静置25min后进行油水分离,得到富含高含量羟基酪醇核桃毛油和果水;
(8)富含高含量羟基酪醇核桃毛油静置,得到富含羟基酪醇的核桃油。
实施例3:制备具有抗疲劳功能的蛋白肽
1、制备蛋白肽
将新鲜金枪鱼肉加入5L纯化水,用高速剪切机剪切30min后转移到提取罐,加入Alcalase2.4酶2g、胰蛋白酶6g,酶解温度35℃;搅拌酶解反应6h;然后用10000rpm的管式离心机离心,得到酶解产物。
将酶解产物离心后的上清液首先用泵入分子量为3000Da的超滤膜进行超滤,过膜温度控制在25℃以下,流速50mL/min,压力15bar,收集透过膜的酶解液。将透过第一滤膜(超滤膜)的酶解液通过分子量为500Da的第二滤膜(纳滤膜)纳滤膜,过膜温度控制在30℃,压力为25bar,将膜***截留的酶解液直接喷雾干燥,得到的奶白的粉末为蛋白肽固体,铝箔袋分装后,置于Co-60辐射灭菌装置灭菌3h,即得蛋白肽成品。
2、蛋白肽提纯
将制备的蛋白肽配置成100mg/ml溶液,通过大孔吸附树脂进行吸附,分别用纯水,10%乙醇,25%乙醇,50%乙醇,75%乙醇和95%乙醇洗脱,得到6个洗脱组分。对6个洗脱组分分别进行一氧化氮合酶活性检测(Nitric Oxide Synthase Activity Assay试剂盒,Colorimetric)。检测结果表明50%乙醇洗脱组分具有最好的活性。
将50%乙醇洗脱组分再次利用HPLC进行分离,分离条件如下:仪器:Agilent1260型高效液相色谱仪,色谱柱:依利特SimoChrom ODS-BP5μm(4.6mm×250mm),流动相:甲醇和0.1%TFA水,流速:1ml/min,检测波长:220nm,收集不同时段的洗脱液(图1),经过一氧化氮合酶活性检测,发现30-32min洗脱液的活性最好。
3、功能蛋白肽的HPLC分析
将30-32min洗脱液用高效液相色谱C18分析柱对具有最好的效果的30%甲醇洗脱峰再次进行纯化,得到更纯的单一峰(图2),使分离的样品达到可以测序的纯度。
4、蛋白肽结构鉴定
对纯化的蛋白肽进行进行多肽结构鉴定,色谱条件如下所示:仪器类型:LCMS-9030Q-TOF四极杆-飞行时间质谱仪,色谱柱:Inertsil HILIC C18反相色谱柱150mm×3.0mm,3μm,岛津,日本),柱温:30℃,进样体积:1.0μL;流动相:A为0.1%(v/v)甲酸水溶液,流动相B为0.1%(v/v)甲酸乙腈溶液,梯度洗脱程序:0-5.0min,5%,5.0-10.0min,10%,10.0-15.0min,20%,15.0-20.0min,40%;流速为0.3mL/min,质谱离子模式:ESI源,采用正离子模式,离子源接口电压:4.0kV,气体流速:10.0L/min,干燥气体流速:10.0L/min,加热气体流速:10.0L/min;加热模块温度:400℃,接口温度:300℃,扫描模式:全扫描模式(m/z50-2000)。
测序获得的蛋白肽的氨基酸序列为YYMEGDFAFGDNY,其分子量为1535.7Da,该多肽命名为PA1。
5、检测蛋白肽抗疲劳的功能
选雄性小鼠体重18-22g,将小鼠分为空白对照组、实验组两组,其中,实验组分为低剂量组、中剂量组和高剂量组,空白对照组、低剂量组、中剂量组和高剂量组均小鼠10只,分别投喂合成的PA1多肽;进行小鼠抗疲劳游泳实验。在小鼠尾部处负重体重5%的铅丝后,投入到水槽中游泳,观察和记录小鼠自入水到沉入水中不能浮出水面(处于力竭状态)的时间,实验水温为25℃,水深25cm。实验结果如表1和图3所示。记录相关数据于表1,并对各组数据进行线条图统计,见图3。
表1:不同剂量PA1多肽对小鼠负重游泳时间的影响表(x±s)
组别 | n | 剂量(mg/kg) | 力竭状态时间(s) |
对照组 | 10 | - | 365.67±51.2 |
低剂量组 | 10 | 50 | 554.59±62.4 |
中剂量组 | 10 | 100 | 677.32±45.7 |
高剂量组 | 10 | 150 | 720.75±74.1 |
实验结果显示,与对照组相比,低剂量组、中剂量组和高剂量组的小鼠力竭状态时间均明显延长(p<0.01),由此,可知PA1蛋白肽能够对小鼠起到抗疲劳的作用,与低剂量组相比,中剂量组和高剂量组有明显的延长,显示PA1多肽的抗疲劳效果具有浓度依赖性,但150mg/kg组相比于100mg/kg组并没有显著的延长负重游泳时间。
实施例4:将蛋白肽添加到枸杞果油和核桃油的组合物
将实施例3中制备的蛋白肽添加到溶剂中,制成具有抗疲劳功能的制品,所述的溶剂包含枸杞果油和核桃油;
其中一种具体的制品的制备过程如下:
1)取枸杞果油2份、核桃油3份、水10份进行混合后搅拌制成溶剂;
2)将蛋白肽添加到溶剂中,添加浓度为20-65μg/ml。
进一步的,所述的溶剂中还添加有用于改善口味的调味料,为甜味剂或酸味剂。
本发明将具有抗疲劳功能的蛋白肽添加到枸杞果油和核桃油制成的溶剂中,使制成的可食用的制品即具有抗视力疲劳的功效,也具有缓解疲劳的效果。
以上实施例仅用以说明本发明的技术方案,而非对其限制;尽管参照前述实施例对本发明进行了详细的说明,本领域的普通技术人员应当理解:其依然可以对前述各实施例所记载的技术方案进行修改,或者对其中部分技术特征进行等同替换;而这些修改或者替换,并不会使相应技术方案的本质脱离本发明技术方案的保护范围。
Claims (10)
1.一种枸杞果油和核桃油的组合物,其特征在于,所述的组合物中包含有任一比例的枸杞果油和核桃油。
2.根据权利要求1所述的一种枸杞果油和核桃油的组合物,其特征在于,所述的组合物中还添加有具有抗疲劳效果的蛋白肽。
3.根据权利要求2所述的一种枸杞果油和核桃油的组合物,其特征在于,所述的具有抗疲劳效果的蛋白肽,是金枪鱼肉的Alcalase2.4酶和胰蛋白酶的酶解液进行离心获得的。
4.根据权利要求3所述的一种枸杞果油和核桃油的组合物,其特征在于,所述的酶解液,其酶解温度为35℃的条件下酶解5-6h。
5.根据权利要求3所述的一种枸杞果油和核桃油的组合物,其特征在于,所述的Alcalase2.4酶和胰蛋白酶的酶活比为1:3。
6.根据权利要求2所述的一种枸杞果油和核桃油的组合物,其特征在于,所述的蛋白肽的分子量为500-3000Da。
7.根据权利要求6所述的一种枸杞果油和核桃油的组合物,其特征在于,所述的蛋白肽,其中一种多肽的氨基酸序列为YYMEGDFAFGDNY。
8.根据权利要求1所述的一种枸杞果油和核桃油的组合物及其应用,其特征在于,所述的组合物中还包含常用的调味料。
9.根据权利要求8所述的一种枸杞果油和核桃油的组合物及其应用,其特征在于,所述的调味料,为甜味剂或酸味剂。
10.权利要求1-9任意一项所述制备方法制备的组合物在抗疲劳制品中的应用。
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