CN115104630B - 一种增加茶树叶片氟富集和抗冻能力的组合物及其制备方法 - Google Patents
一种增加茶树叶片氟富集和抗冻能力的组合物及其制备方法 Download PDFInfo
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Abstract
本发明提出了一种增加茶树叶片氟富集和抗冻能力的组合物、制备方法及应用，属于茶树技术领域，由以下原料制备而成：钙盐3‑5份、复合酶3‑7份、茶细胞壁提取物5‑12份、抗氧化剂1‑2份、水30‑50份、增稠剂2‑3份、抗冻剂1‑2份。通过涂覆本发明组合物后，茶树能提高富集F能力，且对茶树有很好的防冻作用。
Description
技术领域
本发明涉及茶树技术领域，具体涉及一种增加茶树叶片氟富集和抗冻能力的组合物、制备方法及应用。
背景技术
茶树(CamelliasinensisL.O.Kuntze)是有较强富集氟能力的几种植物之一，可吸收、富集分散在大气、水和土壤中的氟。叶片是茶树富集氟的主要器官，因而，饮茶成为人们摄取氟的方式之一。氟是人体所需要的一种微量元素，适量的氟能促进人体骨骼和牙齿的钙化、增加骨骼强度，并促进牙釉质的形成，防治龋齿；但若长期摄入过量氟化物，将干扰人体钙代谢和骨组织中胶原蛋白合成，引起氟中毒，临床表现为氟斑牙、氟骨病和尿氟增高等。世界卫生组织(WHO)规定，人均每天适宜的氟摄入量(Adequateintakes,AI)为2.5-4.0mg。据此，我国农业部规定了茶叶中氟化物含量不得超过200mg/kg。因此，大量的文献记载了茶树富集氟的特点，以及关注如何降低茶叶中的氟。
茶树的产量易受冻害影响，冻害发生时，生长势好的成龄茶树受冻程度轻，幼苗和衰老茶树则易受冻，影响产量，况且随着茶饮品量产以及茶树富集氟的衍生品的出现，茶叶作为待加工提取物，被广泛应用于饮料、含氟牙膏、废水处理、河道治理等技术领域，因此如何增加茶叶中的氟成为亟待解决的新问题。
发明内容
本发明的目的在于提出一种增加茶树叶片氟富集和抗冻能力的组合物、制备方法及应用，通过涂覆本发明组合物后，茶树能提高富集F能力，且对茶树有很好的防冻作用。
本发明的技术方案是这样实现的：
本发明提供一种增加茶树叶片氟富集和抗冻能力的组合物，由以下原料制备而成：钙盐、复合酶、茶细胞壁提取物、抗氧化剂、水、增稠剂、抗冻剂。
作为本发明的进一步改进，由以下原料制备而成：钙盐3-5份、复合酶3-7份、茶细胞壁提取物5-12份、抗氧化剂1-2份、水30-50份、增稠剂2-3份、抗冻剂1-2份。
特别提出，由以下原料制备而成：钙盐4份、复合酶5份、茶细胞壁提取物7份、抗氧化剂1.5份、水40份、增稠剂2.5份、抗冻剂1.5份。
值得考虑地，所述钙盐选自氯化钙、硫酸钙、硝酸钙中的至少一种。
值得考虑地，所述复合酶为果胶酶、蛋白酶的复配混合物，质量比为(4-7)：2，所述蛋白酶选自木瓜蛋白酶、无花果蛋白酶、菠萝蛋白酶中的至少一种。
值得考虑地，所述抗氧化剂选自虾青素、角黄素、叶黄素，β-胡萝卜素、白藜芦醇中的至少一种。
本发明进一步保护一种上述增加茶树叶片氟富集和抗冻能力的组合物的制备方法，其特征在于，包括以下步骤：将钙盐、复合酶、茶细胞壁提取物、抗氧化剂和增稠剂溶于水中，加热至35-40℃搅拌混合均匀，冷却至室温，制得增加茶树叶片氟富集和抗冻能力的组合物，将其均匀涂覆在茶叶表面，可用于治理氟污染或氟含量过高土壤，吸附土壤氟后的茶树可作为高氟提取物的原料，应用于含氟食品、含氟日化产品、及含氟药剂的开发等。
值得考虑地，所述增稠剂选自甲基纤维素、羟丙基甲基纤维素、羧甲基纤维素钠、羟乙基纤维素、淀粉、明胶、干酪素、瓜尔胶、甲壳胺、阿拉伯树胶、黄原胶、大豆蛋白胶、天然橡胶、羊毛脂、琼脂中的至少一种。
作为本发明的进一步改进，所述茶细胞壁提取物的制备方法如下：
将鲜叶加入预冷乙醇溶液匀浆，离心分离沉淀，沉淀加入预冷乙醇溶液，冰浴下浸提50-70min，期间每隔5-15min充分搅拌0.5-1.5min，离心，沉淀依次用冰丙酮，冰甲醇-三氯甲烷溶液和冰甲醇各洗涤1-3次，每次洗涤后，悬浮液离心，弃去上清液，将沉淀物至于通风设备中吹1-3天，使得沉淀物中的甲醛完全挥发干净，在沉淀物中加入水至刚好将沉淀物浸没，冷冻干燥，得到茶细胞壁提取物。
值得考虑地，所述乙醇溶液含量为70-90wt％；鲜叶和乙醇溶液的固液比1：(5-10)g/mL；所述冰甲醇-三氯甲烷的体积比为1:1。
作为本发明的进一步改进，所述抗冻剂由以下原料制备而成：海藻酸钠、聚乙烯醇、氯化钙、水、氯化钠、果胶酶、成膜助剂、分散剂、增稠剂。
作为本发明的进一步改进，所述抗冻剂由以下原料按重量份制备而成：海藻酸钠5-12份、聚乙烯醇2-5份、氯化钙1-2份、水30-50份、氯化钠2-5份、果胶酶2-3份、成膜助剂1-3份、分散剂1-2份、增稠剂2-3份。
作为本发明的进一步改进，所述抗冻剂的制备方法如下：
S1.将果胶酶、海藻酸钠、聚乙烯醇、氯化钠、成膜助剂、分散剂、增稠剂溶于3/4的水中，搅拌混合均匀；
S2.将氯化钙溶于剩余的水中，得到氯化钙水溶液；
S3.边搅拌边向步骤S1中滴加步骤S2制得的氯化钙水溶液，加入后高速乳化，得到茶树抗冻剂。
作为本发明的进一步改进，所述搅拌转速为300-500r/min；所述高速乳化转速为10000-12000r/min，乳化时间为3-5min。
值得考虑地，所述成膜助剂选自丙二醇丁醚、丙二醇甲醚醋酸酯、乙二醇丁醚中的至少一种。
值得考虑地，所述分散剂选自微晶石蜡、硬脂酸钡、硬脂酸锌、硬脂酸钙、硬脂酸镉、硬脂酸镁、硬脂酸铜、聚乙烯蜡、聚乙二醇200、聚乙二醇400中的至少一种。
值得考虑地，所述增稠剂选自甲基纤维素、羟丙基甲基纤维素、羧甲基纤维素钠、羟乙基纤维素、淀粉、明胶、干酪素、瓜尔胶、甲壳胺、阿拉伯树胶、黄原胶、大豆蛋白胶、天然橡胶、羊毛脂、琼脂中的至少一种。
值得考虑地，所述增稠剂为明胶和羟乙基纤维素的复配混合物，质量比为(3-5)：1。
将制得的所述茶树抗冻剂均匀涂覆在茶树茎和幼芽叶子表面即可。
本发明具有如下有益效果：本发明通过茶叶表面涂覆茶细胞壁提取物、钙离子，能渗透入茶叶内，形成氟复合物，不仅降低了单一F-的毒害作用，添加的复合酶和抗氧化剂，还减少氟对细胞器的破坏，从而提高茶树富集氟的能力；另外，本发明制得的组合物喷洒幼芽和嫩叶，能够在茶树幼芽叶子和茎表面形成一层水凝胶薄膜，薄膜可以减少幼苗嫩叶的蒸腾作用，增强茶树抗寒能力，对茶树有很好的防冻作用，水凝胶层富含有水分，使得茶树在有霜的夜间，使气温降低的速度减慢，水凝胶能连续不断的为茶树提供水分直到黎明气温升高时停止，就可以防止茶树叶片温度下降到冰点以下，同时茶树上形成的冰片会在短时间内融化，起到很好的防霜防冻的效果；
本发明添加的复合酶包括果胶酶和蛋白酶，果胶酶能酶解茶树果胶产生小分子寡糖、短糖等，从而在表面形成多糖组合物；蛋白酶能酶解蛋白质得到各种氨基酸、短肽等物质，释放大量的活性基团，能有效与F-络合，形成果糖-F、氨基酸-F复合物等，从而降低F离子的毒害，提高茶树富集氟的能力，特殊季节通过涂覆本发明组合物后，起到很好的保水抗寒效果，具有广阔的应用前景。
附图说明
为了更清楚地说明本发明实施例或现有技术中的技术方案，下面将对实施例或现有技术描述中所需要使用的附图作简单地介绍，显而易见地，下面描述中的附图仅仅是本发明的一些实施例，对于本领域普通技术人员来讲，在不付出创造性劳动性的前提下，还可以根据这些附图获得其他的附图。
图1为本发明测试例1中各处理组氟含量对比图；
图2为本发明测试例2中各处理组相对电导率对比图；
图3为本发明抗冻效果的数据表。
具体实施方式
下面将对本发明实施例中的技术方案进行清楚、完整地描述，显然，所描述的实施例仅仅是本发明一部分实施例，而不是全部的实施例。基于本发明中的实施例，本领域普通技术人员在没有做出创造性劳动前提下所获得的所有其他实施例，都属于本发明保护的范围。
实施例1
组合物的原料组成(重量份)：硫酸钙3份、复合酶3份、茶细胞壁提取物5份、β-胡萝卜素1份、水30份、明胶2份、抗冻剂1份。复合酶为果胶酶、无花果蛋白酶的复配混合物，质量比为7：2。
茶细胞壁提取物的制备方法如下：
将10g鲜叶加入50mL预冷的70wt％的乙醇溶液匀浆，，离心分离沉淀，沉淀加入预冷乙醇溶液，冰浴下浸提50-70min，期间每隔5min充分搅拌0.5min，离心，沉淀依次用冰丙酮，冰甲醇-三氯甲烷溶液(冰甲醇-三氯甲烷的体积比为1:1)和冰甲醇各洗涤1次，每次洗涤后，悬浮液离心，弃去上清液，将沉淀物至于通风设备中吹1天，使得沉淀物中的甲醛完全挥发干净，在沉淀物中加入水至刚好将沉淀物浸没，冷冻干燥，得到茶细胞壁提取物。
抗冻剂的制备方法如下：
原料组成(重量份)：海藻酸钠5份、聚乙烯醇2份、氯化钙1份、水30份、氯化钠2份、果胶酶2份、乙二醇丁醚1份、聚乙二醇200为1份、增稠剂2-3份。增稠剂为明胶和羟乙基纤维素的复配混合物，质量比为3：1，包括以下步骤：
S1.将果胶酶、海藻酸钠、聚乙烯醇、氯化钠、乙二醇丁醚、聚乙二醇200、增稠剂溶于3/4的水中，300r/min搅拌混合均匀；
S2.将氯化钙溶于剩余的水中，得到氯化钙水溶液；
S3.边搅拌边向步骤S1中滴加步骤S2制得的氯化钙水溶液，加入后，10000r/min乳化3min，得到茶树抗冻剂。
增加茶树叶片氟富集和抗冻能力的组合物的制备方法，包括以下步骤：将硫酸钙、复合酶、茶细胞壁提取物、β-胡萝卜素、抗冻剂和明胶溶于水中，加热至40℃搅拌混合均匀，冷却至室温，制得增加茶树叶片氟富集和抗冻能力的组合物，将其均匀涂覆在茶叶表面。
实施例2
组合物原料组成(重量份)：氯化钙4份、复合酶5份、茶细胞壁提取物7份、虾青素1.5份、水40份、羟丙基甲基纤维素2.5份、抗冻剂1.5份。复合酶为果胶酶、菠萝蛋白酶的复配混合物，质量比为5：2。
茶细胞壁提取物的制备方法如下：
将10g鲜叶加入100mL预冷的90wt％的乙醇溶液匀浆，，离心分离沉淀，沉淀加入预冷乙醇溶液，冰浴下浸提60min，期间每隔10min充分搅拌1min，离心，沉淀依次用冰丙酮，冰甲醇-三氯甲烷溶液(冰甲醇-三氯甲烷的体积比为1:1)和冰甲醇各洗涤3次，每次洗涤后，悬浮液离心，弃去上清液，将沉淀物至于通风设备中吹2天，使得沉淀物中的甲醛完全挥发干净，在沉淀物中加入水至刚好将沉淀物浸没，冷冻干燥，得到茶细胞壁提取物。
抗冻剂的制备方法如下：
原料组成(重量份)：海藻酸钠7份、聚乙烯醇4份、氯化钙1.5份、水40份、氯化钠3.5份、果胶酶2.5份、丙二醇甲醚醋酸酯2份、硬脂酸钡1.5份、增稠剂2.5份。增稠剂为明胶和羟乙基纤维素的复配混合物，质量比为4：1，包括以下步骤：
S1.将果胶酶、海藻酸钠、聚乙烯醇、氯化钠、丙二醇甲醚醋酸酯、硬脂酸钡、增稠剂溶于3/4的水中，500r/min搅拌混合均匀；
S2.将氯化钙溶于剩余的水中，得到氯化钙水溶液；
S3.边搅拌边向步骤S1中滴加步骤S2制得的氯化钙水溶液，加入后，12000r/min乳化5min，得到茶树抗冻剂。
增加茶树叶片氟富集和抗冻能力的组合物的制备方法，包括以下步骤：将氯化钙、复合酶、茶细胞壁提取物、虾青素、抗冻剂和羟丙基甲基纤维素溶于水中，加热至37℃搅拌混合均匀，冷却至室温，制得增加茶树叶片氟富集和抗冻能力的组合物，将其均匀涂覆在茶叶表面。
实施例3
与实施例2相比，组合物中未添加氯化钙，其他条件均不改变。
组合物原料组成(重量份)：复合酶9份、茶细胞壁提取物7份、虾青素1.5份、水40份、羟丙基甲基纤维素2.5份。复合酶为果胶酶、菠萝蛋白酶的复配混合物，质量比为5：2。
实施例4
与实施例2相比，组合物中未添加复合酶，其他条件均不改变。
组合物原料组成(重量份)：氯化钙9份、茶细胞壁提取物7份、虾青素1.5份、水40份、羟丙基甲基纤维素2.5份。复合酶为果胶酶、菠萝蛋白酶的复配混合物，质量比为5：2。
测试例1提高茶树富集氟的能力
茶树枝条水培：将1年生和2年生茶树扦插苗剪去根部，枝条使用水培法培养，分别涂覆实施例1-4制得的增加茶树叶片氟富集和抗冻能力的组合物，每天喷施1次，叶面叶背均匀涂覆，以喷施纯水为对照，喷施2星期后，使用含有0.8mM氟化钠的营养液培养茶树枝条1天，分别收集不同处理的茶树叶片，测定叶片氟含量。
结果见图1。注释：*为与对照组相比，P<0.05。
由图1可知，无论是1年生茶树枝条，还是2年生茶树枝条，涂覆本发明实施例制得的增加茶树叶片氟富集和抗冻能力的组合物较对照均显著升高。实施例3、4的效果不如实施例2，可见，单一的复合酶或氯化钙的效果不佳，本发明添加钙/酶复合物，能提高茶树吸收外源氟的能力，可用于治理氟污染或氟含量过高土壤，吸附土壤氟后的茶树可作为高氟提取物的原料，应用于含氟食品、含氟日化产品、及含氟药剂的开发等。果胶酶能酶解茶树果胶产生小分子寡糖、短糖等，蛋白酶能酶解蛋白质得到各种氨基酸、短肽等物质，释放大量的活性基团，能有效与F-络合，形成果糖-F、氨基酸-F复合物等，从而降低F离子的毒害，提高茶树富集氟的能力，具有协同增效的效果。
测试例2提高茶树抗冻的能力
茶树枝条水培：将1年生和2年生茶树扦插苗剪去根部，枝条使用水培法培养，分别涂覆实施例1-4制得的抗冻剂，分别取5mM Ca、10mM Ca、0.05g/ml PE、0.1g/ml PE，每天喷施1次，叶面叶背均匀涂覆，以喷施纯水为对照，喷施2星期后，以常温为对照，对枝条进行-10℃处理，测定叶片电导率，评估枝条抗冻能力。
结果见图2和图3。注释：*为与对照组相比，P<0.05。
涂覆抗冻剂后，常温(NT)条件下不同处理相对电导率含量没有差异；-10℃低温处理2h，所有处理组相对电导率均较常温显著升高，说明茶树受到-10℃的低温伤害；-10℃低温条件下，实施例1、2处理组相对电导率显著低于对照(喷施水)，说明本发明实施例制得的组合物能缓解茶树在低温条件下的叶片受害程度。
实施例3、4与实施例2处理组相比，-10℃低温条件下的相对电导率较高，说明本发明添加的复合酶和氯化钙组合能更好的形成膜层，从而进一步改善茶树的防冻效果，具有协同增效的作用。
以上所述仅为本发明的较佳实施例而已，并不用以限制本发明，凡在本发明的精神和原则之内，所作的任何修改、等同替换、改进等，均应包含在本发明的保护范围之内。
Claims (1)
1.一种增加茶树叶片氟富集和抗冻能力的组合物，其特征在于，由以下重量份的原料制备而成：硫酸钙3份、复合酶3份、茶细胞壁提取物5份、β-胡萝卜素1份、水30份、明胶2份、抗冻剂1份；复合酶为果胶酶、无花果蛋白酶的复配混合物，质量比为7：2；
所述茶细胞壁提取物的制备方法如下：
将10g鲜叶加入50mL预冷的70wt%的乙醇溶液匀浆，离心分离沉淀，沉淀加入预冷乙醇溶液，冰浴下浸提50-70min，期间每隔5min充分搅拌0.5min，离心，沉淀依次用冰丙酮，冰甲醇-三氯甲烷溶液和冰甲醇各洗涤1次，每次洗涤后，悬浮液离心，弃去上清液，将沉淀物置于通风设备中吹1天，使得沉淀物中的甲醛完全挥发干净，在沉淀物中加入水至刚好将沉淀物浸没，冷冻干燥，得到茶细胞壁提取物，其中冰甲醇-三氯甲烷的体积比为1:1；
所述抗冻剂由以下重量份的原料制备而成：海藻酸钠5份、聚乙烯醇2份、氯化钙1份、水30份、氯化钠2份、果胶酶2份、乙二醇丁醚1份、聚乙二醇200为1份、增稠剂2-3份，增稠剂为明胶和羟乙基纤维素的复配混合物，质量比为3：1，所述抗冻剂的制备方法包括以下步骤：
S1.将果胶酶、海藻酸钠、聚乙烯醇、氯化钠、乙二醇丁醚、聚乙二醇200、增稠剂溶于3/4的水中，300r/min搅拌混合均匀；
S2.将氯化钙溶于剩余的水中，得到氯化钙水溶液；
S3.边搅拌边向步骤S1中滴加步骤S2制得的氯化钙水溶液，加入后，10000r/min乳化3min，得到茶树抗冻剂；
增加茶树叶片氟富集和抗冻能力的组合物的制备方法，包括以下步骤：将硫酸钙、复合酶、茶细胞壁提取物、β-胡萝卜素、抗冻剂和明胶溶于水中，加热至40℃搅拌混合均匀，冷却至室温，制得增加茶树叶片氟富集和抗冻能力的组合物，将其均匀涂覆在茶叶表面。
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