CN114041598A - 脂溶性植物化合物提质增效加工方法及应用 - Google Patents
脂溶性植物化合物提质增效加工方法及应用 Download PDFInfo
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- Acyclic And Carbocyclic Compounds In Medicinal Compositions (AREA)
Abstract
本发明公开了一种脂溶性植物化合物提质增效加工方法，属于现代营养食品加工技术领域。本发明以一种纳米淀粉粒子，通过酶促表面扩链‑低温聚集以在内核表面形成结晶外层，得到特定核壳冠结构固态淀粉粒子，并将其作为载体，通过pH驱动自组装技术制备载运植物化学物淀粉复合物，从而提高脂溶性植物化合物的加工特性与营养品质。本发明的产品解决了脂溶性天然植物化合物低溶解度、低稳定性、低载量、低生物利用度等难题，其应用范围可涉及到功能食品、医药、日用化妆品等领域。
Description
技术领域
本发明属于现代营养食品加工技术领域，具体涉及一种脂溶性植物化合物提质增效加工方法。
背景技术
近年来，随着生态环境的恶化，人口老龄化现象日益加剧，各种慢性疾病，例如冠心病，糖尿病，高血压，癌症等疾病发生率不断增加，这一变化加速了人们生活方式的转变，尤其在健康营养饮食方面。已经有研究表明，合理的饮食方式可以有效降低非遗传慢性疾病的患病风险，所以越来越多的消费者倾向于选择不仅能满足基本营养需求还能有益身心健康的食品。由此，开发功能性食品成为当今食品工业发展的必然趋势，其不仅有利于国民的身心健康与社会和谐，对经济发展和社会进步也具有重要意义。
生物活性物质在改善人类健康方面发挥重要作用，它是功能性食品的核心成分，特别地，脂溶性生物活性物质是生物活性物质中的重要组成部分，其具有抗癌、抗炎、抗氧化和免疫调节等多种生理功能，如多不饱和脂肪酸、类胡萝卜素、植物甾醇等，因此，促使了消费者对这类活性物质的需求呈现不断增长的趋势。对于食品工业需要不断开发富含脂溶性活性物质的功能性食品，但由于大部分脂溶性活性物质具有水溶性差、极易氧化降解且生物接收度低等缺点，限制了其作为营养强化剂的应用。现阶段包埋脂溶性活性物质的常见体系有乳液、脂质体、固体脂质纳米颗粒、微胶囊、水凝胶等，但是在不同环境压力下，如在加热、冻融、苛刻pH、高离子强度等加工条件下容易引起体系失稳，降低了活性物质的生物利用度。基于上述原因，提供一种脂溶性植物化合物提质增效加工方法应用前景良好。
发明内容
本发明旨在提供一种脂溶性植物化合物提质增效加工方法，该方法具有操作简便、反应条件可控、绿色和可连续化生产等优势。
[技术构思]本发明以纳米淀粉颗粒为内核物质，先通过酶促反应在内核表面扩链-低温聚集结晶制备特定核壳冠结构固态淀粉粒子，然后通过pH驱动自组装技术制备载运植物化学物淀粉复合物，从而提高脂溶性植物化合物的加工特性与营养品质。
本发明的目的通过下述技术方案来实现：
本发明的第一方面在于提供一种脂溶性植物化合物提质增效加工方法，依次包括如下步骤：
(1)将纳米淀粉颗粒溶解于水，以得到质量百分比浓度为0.1-5％的纳米淀粉溶液；将所述纳米淀粉溶液与10-200U/g底物的特异性转苷酶混匀，并于35-50℃恒温反应3-24h，然后加热灭酶、离心、取沉淀干燥得到固态微纳米级粒子；
(2)取质量比为1:(10-200)的脂溶性植物化合物与微纳米淀粉粒子溶解于0.1-1mol/L的氢氧化钠溶液中，配成微纳米淀粉粒子的质量百分比浓度为1-20％的悬浊液，调节体系pH值至8.0-12.0，置于10-50℃下恒温反应同时持续搅拌10-120min；然后离心、取沉淀物真空干燥，即得到载运植物化学物淀粉复合物。
作为本发明的一种优选实施方式，所述纳米淀粉颗粒的分子量为2.0-10×107g/mol，平均链长DP为10-14，粒径30-100nm。
作为本发明的一种优选实施方式，所述纳米淀粉颗粒来源于植物籽粒胚乳。具体地，所述植物籽粒胚乳包括玉米籽粒胚乳、水稻籽粒胚乳、高粱籽粒胚乳、燕麦籽粒胚乳和大麦籽粒胚乳中的至少一种。
作为本发明的一种优选实施方式，所述特异性转苷酶包括葡聚糖蔗糖酶、淀粉蔗糖酶、淀粉葡糖苷酶、葡聚糖磷酸化酶、麦芽糖转葡糖基酶和葡聚糖分支酶中的至少一种。
作为本发明的一种优选实施方式，所述固态微纳米级粒子的平均链长DP为20-60，结晶度为8-30％。
作为本发明的一种优选实施方式，所述脂溶性植物化合物包括姜黄素、槲皮素、白藜芦醇、β-胡萝卜素、胡椒碱、叶黄素、番茄红素、辣椒素、植物甾醇、茶黄素、花色苷中的至少一种。
本发明的第二方面在于提供由前述方法制得的载运植物化学物淀粉复合物，所述载运植物化学物淀粉复合物中植物化学物的生物有效性相比游离植物化学物提高了150％以上。
本发明的第三方面在于提供前述的载运植物化学物淀粉复合物在功能食品、药品、日用化学品的制备中的应用。
本发明有以下优点：
1)以纳米淀粉颗粒为内核物质，通过酶促表面扩链-低温聚集以在内核表面形成结晶外层，得到特定核壳冠结构固态淀粉粒子；然后利用pH驱动技术的碱性体系同时诱导脂溶性植物化合物和前述特定核壳冠结构纳米淀粉粒子的表面结晶层共同溶解，并调控糖链螺旋结构与脂溶性植物化合物组装复合制备得到载运脂溶性植物化合物-淀粉复合物，从而提高了脂溶性植物化合物的生物有效性、加工特性与营养品质。
2)本发明制备的载运植物化学物淀粉复合物解决了脂溶性天然植物化合物低稳定性、低生物利用度等难题；其中脂溶性天然植物化合物包括姜黄素、槲皮素、白藜芦醇、β-胡萝卜素、胡椒碱、叶黄素、番茄红素、辣椒素、植物甾醇等天然产物一种或多种。
3)本发明制备的载运植物化学物淀粉复合物中植物化学物生物有效性相比游离植物化学物提高了150％以上。
4)本发明制备的固态微纳米级粒子的平均链长DP为20-60，结晶度为8-30％。
5)本发明充分利用我国资源丰富的作物资源，通过现代绿色生产技术开发本产品，提高了其附加值，为其综合开发利用提供一定的依据。
6)本发明步骤简便，易于操作，反应条件可控，通过酶促反应联合pH驱动自组装技术制备载运植物化学物淀粉复合物，成本相对较低，而且采用清洁生产工艺，对环境基本无污染。
7)本发明制备的载运脂溶性植物化合物-淀粉复合物产品可应用于功能食品、医药、日用化学品等多个领域，市场前景十分看好，经济效益广阔。
附图说明
图1实施例1制备的载运植物化学物淀粉复合物的电镜照片；
图2实施例1制备的纳米淀粉颗粒(A)与固态微纳米级粒子(B)的链长分布图。
具体实施方式
下面结合实施例进一步阐明本发明的内容，但本发明所保护的内容不仅仅局限于下面的实施例。
绝对分子量测定：采用高效液相体积排阻色谱、多角度激光光散射检测器和示差折光检测器联用系统，选用Shodx Ohpak SB-805HQ凝胶色谱柱，0.1mol/L硝酸钠溶液为流动相，流速设为0.7mL/min，折光指数设定为dn/dc＝0.138。
颗粒尺寸测定：采用激光粒度分析仪测定，以波长658nm固定激光为光源，散射光角度为90°，操作温度为25℃。
结晶度测定：采用X射线衍射仪分析，设定功率1600W，铜靶Cu Kα(λ＝1.5406nm)，采用NaI晶体闪烁计数器测量其强度，扫描速度为4°/min，扫描范围3～40°。
链长分布测定：采用高效阴离子交换色谱与安培脉冲检测器联用系统测定链长分布情况。选用CarboPacPA 200离子交换柱，并设定梯度洗脱，其流动相分别是(A)：0.25mol/L的NaOH溶液；(B)1.0mol/L的NaAC溶液；超纯水。
生物有效性测定：采用Englyst体外模拟消化模型(Englyst,H.N.,Kingman,S.M.,&Cummings,J.H.(1992).Classification and measurement of nutritionallyimportant starch fractions.European Journal of Clinical Nutrition,46,30-50)，监测载运植物化学物淀粉复合物在经过模拟胃肠道消化后植物化学物(脂溶性植物化合物)释放率，计算公式如下：生物有效性(％)＝CM/CD×100％,式中CM表示消化胶束中植物化学物含量，CD表示植物化学物淀粉复合物中植物化学物含量。
实施例中涉及的生物材料的来源：
特异性转苷酶的来源：来源于葡聚糖蔗糖酶、淀粉蔗糖酶、淀粉葡糖苷酶、葡聚糖磷酸化酶、麦芽糖转葡糖基酶和葡聚糖分支酶，采用从自然界中筛选获得古生菌或细菌经过活化培养、发酵产酶、收集菌体、冻干粉碎等步骤得到酶制剂或者Sigma公司商业化产品。
纳米淀粉颗粒的来源：来源于植物籽粒胚乳，包括玉米、水稻、高粱、燕麦、大麦等，主要通过植物籽粒浸泡、粉碎、离心、分级、干燥等步骤达到纳米淀粉颗粒，其分子量为(2.0-10)×107g/mol，平均链长DP为10-14，粒径30-100nm。
实施例1：
将玉米纳米淀粉颗粒(分子量7.3×107g/mol，平均链长DP为12.5，平均粒径84nm)溶解于纯净水配成质量百分比浓度1％玉米纳米淀粉溶液，添加60U/g底物的特异性葡聚糖蔗糖酶在35℃恒温反应6h，然后加热灭酶、离心、取沉淀物干燥处理，得到玉米固态微纳米级粒子；按照质量比1:50称取脂溶性姜黄素与玉米固态微纳米级粒子共混均匀，溶解于0.4mol/L的氢氧化钠溶液配成质量百分比浓度3％悬浊液，调节体系pH值至8.5，置于10℃下不断搅拌恒温反应30min后离心处理，将所获沉淀物真空干燥，即得载运姜黄素淀粉复合物。
经过测定，玉米固态微纳米级粒子的平均链长DP为39，结晶度21％，载运姜黄素淀粉复合物中姜黄素的生物有效性相比原脂溶性姜黄素(游离态脂溶性姜黄素)提高了348％。
实施例2：
将高粱纳米淀粉颗粒(分子量为3.9×107g/mol，平均链长DP为11.6，平均粒径68nm)溶解于纯净水配成质量百分比浓度0.2％高粱纳米淀粉溶液，添加100U/g底物的特异性淀粉蔗糖酶在50℃恒温反应12h后加热灭酶、离心和干燥处理得到高粱固态微纳米级粒子；按照质量比1:20称取脂溶性β-胡萝卜素与高粱微纳米淀粉粒子共混均匀，溶解于0.1mol/L的氢氧化钠溶液配成质量百分比浓度10％悬浊液，调节体系pH值至10.0，置于20℃不断搅拌恒温反应20min后离心处理，将所获沉淀物真空干燥，即得到载运脂溶性β-胡萝卜素淀粉复合物。
经过测定，高粱固态微纳米级粒子的平均链长DP为52，结晶度25％，载运脂溶性β-胡萝卜素淀粉复合物中β-胡萝卜素的生物有效性相比原脂溶性β-胡萝卜素提高了195％。
实施例3：
将水稻纳米淀粉颗粒(分子量为5.2×107g/mol，平均链长DP为10.9，平均粒径52nm)溶解于纯净水配成质量百分比浓度5％水稻纳米淀粉溶液，添加200U/g底物的特异性葡聚糖磷酸化酶在35℃恒温反应24h后加热灭酶、离心和干燥处理得到水稻固态微纳米级粒子；按照质量比1:150称取脂溶性白藜芦醇与水稻微纳米淀粉粒子共混均匀，溶解于1mol/L的氢氧化钠溶液配成质量百分比浓度1％悬浊液，调节体系pH值至12.0，置于50℃不断搅拌恒温反应10min后离心处理，将所获沉淀物真空干燥，即得到载运白藜芦醇淀粉复合物。
经过测定，水稻固态微纳米级粒子的平均链长DP为45，结晶度23％，载运白藜芦醇淀粉复合物中白藜芦醇生物有效性相比原脂溶性白藜芦醇提高了268％。
实施例4
参照实施例2，区别仅在于，将脂溶性β-胡萝卜素替换为姜黄素、槲皮素、白藜芦醇、胡椒碱、叶黄素、番茄红素、辣椒素、茶黄素、花色苷和植物甾醇中的任一种多种，控制脂溶性植物化合物的总量与高粱微纳米淀粉粒子的质量比为1:(10-200)，其他条件不变，得到对应的载运脂溶性植物化合物淀粉复合物。
经过测定，脂溶性植物化合物淀粉复合物中各植物化合物的生物有效性相比原植物化合物提高了150％以上。
实施例5：
参照实施例1，区别仅在于，将特异性葡聚糖蔗糖酶用量由60U/g底物分别替换为5U/g底物、10U/g底物、200U/g底物、1000U/g底物，其他条件不变，得到对应的载运姜黄素淀粉复合物。所得淀粉复合物的性能结果见表1。
表1不同特异性葡聚糖蔗糖酶用量所得载运姜黄素淀粉复合物结果
对比例1：
参照实施例1，区别仅在于，采用普通玉米淀粉(分子量为7.5×108g/mol，平均链长DP为70.1，平均粒径11.6μm)替换玉米固态微纳米级粒子，其他条件不变，得到对应的载运姜黄素淀粉复合物。
经测定，对比例1中的普通玉米淀粉粒子的平均链长DP为70.1，结晶度34％；载运姜黄素淀粉复合物中姜黄素的生物有效性相比原姜黄素仅提高了47％。
对比例2：
参照实施例1，区别仅在于，未添加60U/g底物的特异性葡聚糖蔗糖酶，其他条件不变，得到对应的载运姜黄素淀粉复合物。
经测定，对比例2所得的玉米固态微纳米级粒子的平均链长DP为12.4，结晶度0.2％，载运姜黄素淀粉复合物中姜黄素的生物有效性相比原姜黄素提高了97％。
对比例3：
参照实施例1，区别仅在于，将脂溶性姜黄素与玉米固态微纳米级粒子共混体系的pH由8.5调整至6.5或者13.0，保持其他条件不变，得到对应的载运姜黄素淀粉复合物。所得淀粉复合物的性能结果见表1。
表2不同共混体系的pH值所得载运姜黄素淀粉复合物结果
| 体系pH值 | 平均链长 | 结晶度 | 提高生物有效性 |
| 6.5 | 36 | 14 | 54％ |
| 8.5 | 39 | 21 | 348％ |
| 13.0 | 27 | 12 | 121％ |
实施例6：
根据糖尿病人对特定天然植物化合物的特殊需求，以茶黄素、花色苷等脂溶性植物化合物为原料，设计创制糖尿病人群专用食品新产品，通过原料学评价筛选结合特定核壳冠结构固态微纳米级淀粉粒子的pH驱动自组装技术制备载运植物化学物淀粉复合物，不仅提高了天然植物化合物含量和氧化稳定性，还能改善脂溶性植物化合物的溶解性、生物利用率和不良风味。
本文所描述的具体实施案例仅作为对本发明精神和部分实验做举例说明。本发明所属领域的技术人员可以对所描述的具体实施案例做出各种各样的修改或补充或采用类似的方式替代，但并不会偏离本发明的精神或者超越所附权利要求书所定义的范围。
Claims (9)
1.一种脂溶性植物化合物提质增效加工方法，其特征在于，依次包括如下步骤：
(1)将纳米淀粉颗粒溶解于水，以得到质量百分比浓度为0.1-5％的纳米淀粉溶液；将所述纳米淀粉溶液与10-200U/g底物的特异性转苷酶混匀，并于35-50℃恒温反应3-24h，然后加热灭酶、离心、取沉淀干燥得到固态微纳米级粒子；
(2)取质量比为1:(10-200)的脂溶性植物化合物与微纳米淀粉粒子溶解于0.1-1mol/L的氢氧化钠溶液中，配成微纳米淀粉粒子的质量百分比浓度为1-20％的悬浊液，调节体系pH值至8.0-12.0，置于10-50℃下恒温反应同时持续搅拌10-120min；然后离心、取沉淀物真空干燥，即得到载运植物化学物淀粉复合物。
2.根据权利要求1所述的加工方法，其特征在于，所述纳米淀粉颗粒的分子量为(2.0-10)×107g/mol，平均链长DP为10-14，粒径30-100nm。
3.根据权利要求1所述的加工方法，其特征在于，所述纳米淀粉颗粒来源于植物籽粒胚乳。
4.根据权利要求1所述的加工方法，其特征在于，所述植物籽粒胚乳包括玉米籽粒胚乳、水稻籽粒胚乳、高粱籽粒胚乳、燕麦籽粒胚乳和大麦籽粒胚乳中的至少一种。
5.根据权利要求1所述的加工方法，其特征在于，所述特异性转苷酶包括葡聚糖蔗糖酶、淀粉蔗糖酶、淀粉葡糖苷酶、葡聚糖磷酸化酶、麦芽糖转葡糖基酶和葡聚糖分支酶中的至少一种。
6.根据权利要求1所述的加工方法，其特征在于，所述固态微纳米级粒子的平均链长DP为20-60，结晶度为8-30％。
7.根据权利要求1所述的加工方法，其特征在于，所述脂溶性植物化合物包括姜黄素、槲皮素、白藜芦醇、β-胡萝卜素、胡椒碱、叶黄素、番茄红素、辣椒素、植物甾醇、茶黄素、花色苷中的至少一种。
8.由权利要求1-7任一项所述的方法制得的载运植物化学物淀粉复合物，其特征在于，所述载运植物化学物淀粉复合物中植物化学物的生物有效性相比游离植物化学物提高了150％以上。
9.权利要求8所述的载运植物化学物淀粉复合物在功能食品、药品、日用化学品的制备中的应用。
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