CN106690326B - 一种复合海藻多糖降血脂口服液及其制备方法 - Google Patents
一种复合海藻多糖降血脂口服液及其制备方法 Download PDFInfo
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- CN106690326B CN106690326B CN201611128959.9A CN201611128959A CN106690326B CN 106690326 B CN106690326 B CN 106690326B CN 201611128959 A CN201611128959 A CN 201611128959A CN 106690326 B CN106690326 B CN 106690326B
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- blood fat
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Abstract
本发明公开了一种复合海藻多糖降血脂口服液及其制备方法。按质量百分比，该复合海藻多糖降血脂口服液，包括如下组分：海藻多糖提取液60‑90%，山楂提取液1‑10%，荷叶提取液1‑10%，银杏叶提取液1‑10%，桑叶提取液1‑10%，甜味剂0.01‑10%；所述海藻多糖提取液通过高压蒸汽提取得到。按所述质量百分比将各组分进行混合，将混合得到的混合物进行冷藏后，离心取上清液，灌装、封盖、灭菌，得到所述复合海藻多糖降血脂口服液。本发明将具有降血脂功效的多糖和多酚类物质进行协同复配，制备方法步骤简单，利用海藻多糖与多酚的协同增效作用增强了海藻多糖的降血脂功效，达到增加最终产品降血脂功效的目的。
Description
技术领域
本发明涉及海藻多糖的加工利用技术领域，具体涉及一种复合海藻多糖降血脂口服液及其制备方法。
背景技术
大量研究表明，海藻多糖含有硫酸基、糖醛酸等特殊基团，因此具有多种特殊生理功能，如抗病毒、抗凝血、降血糖、降血脂等活性。海藻类多糖由于其独特的生物活性，在功能食品开发上具有巨大潜力。但其分子量大，大多凝胶性强或者粘度高，致使其在临床应用中受到一定限制。针对高分子海藻多糖物质的分子量过大、粘度高、溶解性差等难题，可通过降解高分子海藻多糖，获得寡糖或小分子多糖片段，以提高其穿越机体屏障的灵活性，从而更好地实现生物活性。
目前降解多糖的常用手段包括：酸降解法、氧化降解法和酶解法。中国专利(ZL03140259.3)报道了以羊栖菜或海蒿子等海藻为原料通过酶解、醇沉、碱提、中和等工艺制备降脂减肥的保健品的方法。中国专利(ZL 200410083685.7)通过抗坏血酸和过氧化氢降解海藻硫酸多糖得到分子量为4-100KDa的低分子量海藻硫酸多糖。中国专利(ZL201110343580.0)公布了以有机酸溶液或有机-无机混酸溶液对褐藻原料进行预处理，再通过中和、水提、分级醇沉等工艺提取活性多糖的方法。但酸降解法需要引入磷酸、盐酸和柠檬酸等无机酸或有机酸，产物成分复杂，纯化成本高；而氧化降解法和酶解法的生产成本高，工艺条件复杂难控，反应稳定性和重复性差，不利于工业生产。而高温高压降解法生产成本低，工艺条件简单，不引入其他成分，生产过程绿色环保，适合规模化生产应用。
通过高压蒸汽提取、超滤、浓缩、醇沉等工艺制备出海藻低分子多糖，再复配具有降血脂功能的药食同源中草药提取液，制备出降血脂功效突出的功能口服液。充分利用了海藻多糖与多酚的复配体系在降血脂功能的协同增效作用，同时大分子的海藻多糖通过高压蒸汽提取后降解成容易被人体吸收的小分子海藻多糖和低聚糖，而小分子的海藻多糖和低聚糖不仅具有降血脂功效，也是能够促进肠道益生菌增殖益生元，维持正常的肠道微生态平衡，而且正常的肠道益生菌群还能够调控血脂代谢并抑制血脂过度升高。
本发明通过体内体外实验，证明复合海藻多糖口服液具有显著的降血脂功效。体外具有较强的胆酸盐结合能力，体内能够降低血清胆固醇、甘油三酯和低密度脂蛋白水平，并提高血清高密度脂蛋白水平，同时通过对比实验，证明复配后的海藻多糖口服液比单一的海藻多糖和中草药水提物具有更好的降血脂活性。
发明内容
本发明提供的一种复合海藻多糖口服液，利用高压蒸汽从海藻原料中提取小分子海藻多糖和低聚糖，并复配具有降血脂功效的中草药多酚，得到复合海藻多糖口服液。
一种复合海藻多糖降血脂口服液，按质量百分比，包括如下组分：
进一步地，所述复合海藻多糖降血脂口服液中的海藻多糖含量为0.1-5.4mg/mL，总酚含量为100-500mg/L，pH＝4.0～6.0。
进一步地，所述海藻多糖提取液中的总糖含量为2-6mg/mL，还原糖含量为0.5-1.0mg/mL，pH＝6.0-6.5。
进一步地，所述海藻多糖提取液通过采用高压蒸汽提取方法从海藻原料中提取得到，主要成分包括海藻多糖和低聚糖。
更进一步地，所述海藻原料包括褐藻门中常见的经济藻类，包括海带、昆布、裙带菜、马尾藻、墨角藻和巨藻中的一种以上。
更进一步地，采用高压蒸汽提取方法从海藻原料中提取海藻多糖提取液，包括如下步骤：
(1)将海藻原料洗净、烘干、粉碎，得到海藻原料粉末；
(2)将海藻原料粉末和水混合后，采用高压蒸汽加热提取，将提取液进行离心分离，得到的上清液用截留分子量为500K超滤膜进行分离，收集分子量在500KDa以下的组分；
(3)将步骤(2)收集的组分进行浓缩后，加入食用酒精并冷藏，离心分离得到沉淀物，加水溶解并减压蒸发脱去酒精，得到所述海藻多糖提取液。
优选的，步骤(2)中，所述海藻原料粉末与水的混合质量比为1:20-50。
优选的，步骤(2)中，所述高压蒸汽是在100kPa下，100-125℃的蒸汽。
优选的，步骤(2)中，所述提取的时间为0.5～2小时。
优选的，步骤(3)中，所述浓缩是浓缩至固形物含量为4～10％。
优选的，步骤(3)中，加入食用酒精至酒精浓度为60～90vol％。
优选的，步骤(3)中，所述冷藏是在4℃下冷藏12小时。
优选的，步骤(3)中，所述加水溶解是按照沉淀物与水的质量比1:10-30添加水。
进一步地，所述山楂提取液、荷叶提取液、银杏叶提取液和桑叶提取液通过如下步骤提取得到：
将烘干后的山楂果实、荷叶、银杏叶和荷叶分别加入水中，加热提取，再分别用纱布过滤，得到所述山楂提取液、荷叶提取液、银杏叶提取液和桑叶提取液。
更进一步地，所述烘干后的山楂果实、荷叶、银杏叶和荷叶与水的料液比为1:5-30g/mL。
更进一步地，所述提取的是在80-100℃下加热提取2h。
进一步地，所述甜味剂包括白砂糖、木糖醇、低聚果糖、低聚木糖、低聚异麦芽糖、大豆低聚糖、低聚半乳糖、蜂蜜、阿斯巴甜、三氯蔗糖和甜菊糖苷中的一种以上。
上述任一所述所述的一种复合海藻多糖降血脂口服液的制备方法，包括如下步骤：
按所述质量百分比将各组分进行混合，将混合得到的混合物进行冷藏后，离心取上清液，灌装、封盖、灭菌，得到所述复合海藻多糖降血脂口服液。
进一步地，所述冷藏是在4℃保存12小时。
与现有技术相比，本发明具有如下优点和有益效果：
(1)本发明所采用的海藻多糖提取方法操作简单，不需引入有机酸、酶或强氧化剂，也能达到较好的大分子多糖的降解效果；
(2)本发明通过将具有降血脂功效的多糖和多酚类物质进行协同复配，制备方法步骤简单，利用海藻多糖与多酚的协同增效作用增强了海藻多糖的降血脂功效，比单一的海藻多糖或中草药水提取物具有更好的降血脂活性，达到增加最终产品降血脂功效的目的；
(3)本发明提供的纯天然药食同源的降血脂功能口服液，药食同源的原料安全无副作用，具有显著的降血脂功效，能够降低血清胆固醇、甘油三酯和低密度脂蛋白水平，并提高血清高密度脂蛋白水平，为高血脂患者提供了一种良好的、安全无副作用的辅助治疗高血脂的功能食品；
(4)本发明采用高压蒸汽提取能显著提高海带多糖的提取率，且分子量在500KDa以下的海带多糖具有显著的胆酸盐结合能力；
(5)本发明复合海藻多糖降血脂口服液通过复配后掩盖了海藻原有的腥味和甘味，略带特征草药气味，口感清爽微甜，风味更易被各类人群接受。
附图说明
图1为实施例中不同提取方法的提取率结果对比图。
图2为采用高效凝胶渗透色谱仪(Waters 1525)检测海带粗多糖分子量的分布图。
图3为实施例中三个组分海带粗多糖的体外胆酸盐结合能力对比图。
具体实施方式
以下结合实施例对本发明作进一步阐述，但本发明不限于以下实施例。
实施例1
(1)取1斤市售干海带洗净，烘干并粉碎得到200g海带粉；将海带粉和水按质量比1：20混合，于100kPa下用125℃蒸汽加热提取0.5小时，8000rpm离心分离10分钟，得到上清液；将上清液用截留分子量为500K超滤膜分离，收集分子量在500KDa以下的组分；
(2)将收集的组分浓缩至固形物含量为4％，浓缩液加食用酒精至酒精最终浓度为60vol％，4℃冷藏12小时，离心分离得到沉淀，按1:10加水溶解并减压蒸发除去酒精，得到最终多糖含量为6.0mg/mL，还原糖含量为1.0mg/mL，pH＝6.5的海藻多糖提取液；
(3)取烘干后的山楂果实、荷叶、银杏叶和桑叶分别以1：5的料水比加水于80℃热水提取2h后，纱布过滤得山楂、荷叶、银杏叶和桑叶提取液；
(4)按以下质量百分比，混合各组分：海藻多糖提取液90％，山楂提取液1％，荷叶提取液1％，银杏叶提取液1％，桑叶提取液1％，低聚木糖3％，蜂蜜1％，白砂糖2％；将混合物于4℃冷藏保存12小时，8000rpm离心分离10分钟，取上清液，然后灌装、封盖、灭菌，得最终复合海藻多糖降血脂口服液。
实施例2
(1)取1斤市售干裙带菜洗净，烘干并粉碎得到200g裙带菜粉；将裙带菜粉和水按质量比1：50混合，于100kPa下用100℃蒸汽加热提取2小时，8000rpm离心分离10分钟，得到上清液；将上清液用截留分子量为500K超滤膜分离，收集分子量在500KDa以下的组分；
(2)将收集的组分浓缩至固形物含量为10％，浓缩液加食用酒精至酒精最终浓度为90vol％，4℃冷藏12小时，离心分离得到沉淀，按1:30加水溶解并减压蒸发除去酒精，得到最终多糖含量为2.0mg/mL，还原糖含量为0.5mg/mL，pH＝6.0的海藻多糖提取液；
(3)取烘干后的山楂果实、荷叶、银杏叶和桑叶分别以1：30的料水比加水于100℃热水提取2h后，纱布过滤得山楂、荷叶、银杏叶和桑叶提取液；
(4)按以下质量百分比，混合各组分：海藻多糖提取液：60％，山楂提取液：10％，荷叶提取液：10％，银杏叶提取液10％，桑叶提取液5％，低聚木糖3％，蜂蜜1％，白砂糖1％；将混合物于4℃冷藏保存12小时，8000rpm离心分离10分钟，取上清液，然后灌装、封盖、灭菌，得最终复合海藻多糖降血脂口服液。
实施例3
(1)取1斤市售干昆布洗净，烘干并粉碎得到200g昆布粉；将昆布粉和水按质量比1：40混合，于100kPa下用110℃蒸汽加热提取1小时，8000rpm离心分离10分钟，得到上清液；将上清液用截留分子量为500K超滤膜分离，收集分子量在500KDa以下的组分；
(2)将收集的组分浓缩至固形物含量为6％，浓缩液加食用酒精至酒精最终浓度为80vol％，4℃冷藏12小时，离心分离得到沉淀，按1:20加水溶解并减压蒸发除去酒精，得到最终多糖含量为4.0mg/mL，还原糖含量为0.8mg/mL，pH＝6.2的海藻多糖提取液；
(3)取烘干后的山楂果实、荷叶、银杏叶和桑叶分别以1：20的料水比加水于90℃热水提取2h后，纱布过滤得山楂、荷叶、银杏叶和桑叶提取液；
(4)按以下质量百分比，混合各组分：海藻多糖提取液60％，山楂提取液5％，荷叶提取液10％，银杏叶提取液10％，桑叶提取液10％，大豆低聚糖3％，低聚果糖1％，低聚半乳糖1％；将混合物于4℃冷藏保存12小时，8000rpm离心分离10分钟，取上清液，然后灌装、封盖、灭菌，得最终复合海藻多糖降血脂口服液。
实施例4
海带多糖不同提取方法的提取率比较实验
不同提取方法如表1所列，本实验分别考察了酶辅助提取、超声波辅助提取、热水提取、酸辅助提取和高压蒸汽提取的海带多糖提取率。
取市售干海带洗净，烘干并粉碎得到海带粉；取等量海带粉，按表1所示，用不同提取方法加水提取后，8000rpm离心分离10分钟，得到上清液；将上清液浓缩至固形物含量为5％，加食用酒精至酒精最终浓度为80vol％，4℃醇沉12小时，离心分离取沉淀，冷冻干燥后得海带粗多糖，称重计算提取率。
提取率记为每克海带粉所得海带粗多糖的质量(g/g)。实验数据采用SPSS软件进行统计学分析，所有数据用x±s表示，组间采用t检验，P<0.05为差异具有统计学意义。
表1不同提取方法
不同提取方法的提取率结果如图1所示，由图1可知，高压蒸汽提取法和酸辅助提取法的多糖提取率是最高的，与酶辅助提取法、超声波辅助提取法和热水提取法的多糖提取率具有显著差异性。虽然高压蒸汽提取法和酸辅助提取法两者之间没有显著差异，但酸提法引入杂质，生产成本高，工艺条件复杂难控，不利于工业生产。而高压蒸汽提取法在能达到与酸辅助提取法同样较高的提取率时，能避免引入杂质，简化生产工艺条件，所以用于工业生产具有优势。
实施例5
取1斤市售干海带洗净，烘干并粉碎得到200g海带粉；将海带粉和水按质量比1:50混合，于100kPa下用125℃蒸汽加热提取0.5小时，8000rpm离心分离10分钟，得到上清液；将上清液浓缩至固形物含量为5％，加食用酒精至酒精最终浓度为80vol％，4℃冷藏12小时，离心分离取沉淀，冷冻干燥后得海带粗多糖。
采用高效凝胶渗透色谱仪(Waters 1525)检测海带粗多糖分子量分布，结果如图2所示，由图2可知，海带多糖峰位分子量为566KDa，重均分子量815KDa，因此选用截留分子量为500K和1000K的超滤膜分离海带提取液。
将提取液经过8000rpm离心10分钟所得的上清液用截留分子量为500K和1000K的超滤膜分离，分别收集分子量在500K以下、500K-1000K和1000K以上的三个组分，分别浓缩至固形物含量为5％，加食用酒精至酒精最终浓度为80vol％，4℃冷藏12小时，离心分离取沉淀，冷冻干燥后得三个组分的海带粗多糖：HP-1(<500K)、HP-2(500K～1000K)、HP-3(>1000K)。
测定三个组分海带粗多糖的体外胆酸盐结合能力，以考来烯胺为阳性对照，降血脂药物—考来烯胺(消胆胺)在体内与体外均表现出结合胆酸盐的能力，以纤维素为阴性对照。具体实验步骤如下：
10mg样品加入1ml 0.01M HCl并在37℃消化2小时，再用0.1ml 0.1M NaOH调整pH值到7.0，加5ml含0.5μM牛磺胆酸钠的猪胰酶液(10mg/ml)，模拟肠道消化3小时；消化液离心(8000rpm)20分钟，取上清液，利用固相微萃取分离胆酸盐。采用反相高效液相法测定胆酸盐含量，Waters C18色谱柱(250×4.6mm,5μm)；流动相为甲醇-水(75:25),流速:1.0mLmin；柱温30℃；采用蒸发光散射检测器，漂移管温度为110℃，氮气流速2.8L/min，进样量20μL。胆酸盐结合量记为每100mg样品结合胆酸盐的量(μmol/100mg)，实验数据采用SPSS软件进行统计学分析，组间采用t检验，P<0.05为差异具有统计学意义。
测定结果如图3所示，由图3可知，胆酸盐结合能力为HP-1＞HP-2＞HP-3，高压蒸汽提取法所得海带粗多糖中，分子量小于500KDa的组分具有更好的降血脂活性。因此，选用截留分子量为500K的超滤膜分离，收集分子量在500KDa以下的有效片段制备海藻多糖降血脂口服液。
实施例6
本发明复合海藻多糖降血脂口服液降血脂动物实验。
本实验中所用高脂饲料配方，按质量百分比，包括：大鼠普通基础饲料68.2％，胆固醇1％，胆酸钠0.8％，白糖15％，蛋黄粉5％和猪油10％。将原料充分搅拌混匀，压制成合格的大鼠颗粒饲料，灭菌后存用。
本实验中复合海藻多糖口服液为实施例1、2和3中制得的最终复合海带多糖口服液，其中单海藻多糖口服液和单中药提取液为实施例1中提取的海带多糖提取液和中草药复合提取液；阳性对照组灌胃药物为辛伐他汀。
实验前将80只大鼠(180～200g)在试验环境下饲喂基础饲料，保持正常昼夜循环，自由饮水摄食，1周适应性喂养，室温控制在25℃，相对湿度控制在45％～65％。1周后将大鼠随机分为2组：高血脂组70只，高脂饲料喂养；血脂正常组10只，以正常饲料喂养。两组均自由摄食饮水。造模3周，禁食12h后尾静脉采血，通过日立7180型全自动生化分析仪，采用氧化酶法测其血脂四项指标(总胆固醇TC、甘油三酯TG、高密度脂蛋白HDL-C、低密度脂蛋白LDL-C)。高脂饲料喂养的大鼠血脂四项含量均显著高于正常组大鼠，说明造模成功，并用于实验。
取造模成功的高血脂大鼠70只随机分为7组，即高血脂模型组、单海藻多糖组[1ml/(kg·d)]、单中药提取液组[1ml/(kg·d)]、复合海藻多糖组1[1ml/(kg·d)]、复合海藻多糖组2[1ml/(kg·d)]、复合海藻多糖组3[1ml/(kg·d)]、阳性对照组[辛伐他汀0.33mg/(kg·d)]。分组后，于每日上午灌胃一次，持续4周。血脂正常组用生理盐水灌胃并给予基础饲料，同时高血脂模型组、单海藻多糖组、单中药提取液组、复合海藻多糖组1、复合海藻多糖组2、复合海藻多糖组3、阳性对照组继续给予高脂饲料，保持正常昼夜循环和自由饮水摄食。4周后，禁食12h后尾静脉采血，制成血清，测血脂四项(TC、TG、HDL-C、LDL-C)含量。实验数据采用SPSS软件进行统计学分析，所有数据用x±s表示，组间采用t检验，P<0.05为差异具有统计学意义。
实验结果如表2所示，由表2可知，复合海藻多糖口服液能显著降低高血脂大鼠的血清总胆固醇、甘油三酯和低密度脂蛋白胆固醇水平，显著升高血清高密度脂蛋白胆固醇水平，证明本发明复合海藻多糖口服液具有显著的降血脂功效。同时与单海带多糖组或单中药提取液组比较，复合海藻多糖口服液的降血脂功效更优越，证明复配后的海藻多糖口服液比单一的海藻多糖和中草药水提物具有更好的降血脂活性。
表2本发明复合海藻多糖口服液对高脂大鼠的血清血脂含量的影响
与正常组相比：#p<0.05为差异显著；与模型组相比：*p<0.05为差异显著。
Claims (6)
1.一种复合海藻多糖降血脂口服液，其特征在于，按质量百分比，包括如下组分：
海藻多糖提取液 60-90%，
山楂提取液 1-10%，
荷叶提取液 1-10%，
银杏叶提取液 1-10%，
桑叶提取液 1-10%，
甜味剂 0.01-10%；
所述海藻多糖提取液通过采用高压蒸汽提取方法从海藻原料中提取得到，主要成分包括海藻多糖和低聚糖；所述海藻原料包括褐藻门中常见的经济藻类，包括海带、昆布、裙带菜、马尾藻、墨角藻和巨藻中的一种以上；采用高压蒸汽提取方法从海藻原料中提取海藻多糖提取液，包括如下步骤：
（1）将海藻原料洗净、烘干、粉碎，得到海藻原料粉末；
（2）将海藻原料粉末和水混合后，采用高压蒸汽加热提取，将提取液进行离心分离，得到的上清液用截留分子量为500K超滤膜进行分离，收集分子量在500KDa以下的组分；所述海藻原料粉末与水的混合质量比为1:20-50；所述高压蒸汽是在100kPa下，100-125℃的蒸汽；所述提取的时间为0.5~2小时；
（3）将步骤（2）收集的组分进行浓缩后，加入食用酒精并冷藏，离心分离得到沉淀物，加水溶解并减压蒸发脱去酒精，得到所述海藻多糖提取液；
所述山楂提取液、荷叶提取液、银杏叶提取液和桑叶提取液通过如下步骤提取得到：
将烘干后的山楂果实、荷叶、银杏叶和桑叶分别以1:5-30g/mL的料水比加入水中，在80-100℃下加热提取2h，分别用纱布过滤，得到所述山楂提取液、荷叶提取液、银杏叶提取液和桑叶提取液。
2.根据权利要求1所述的一种复合海藻多糖降血脂口服液，其特征在于，所述复合海藻多糖降血脂口服液中的海藻多糖含量为0.1-5.4mg/mL，总酚含量为100-500mg/L，pH=4.0~6.0。
3.根据权利要求1所述的一种复合海藻多糖降血脂口服液，其特征在于，所述海藻多糖提取液中的总糖含量为2-6mg/mL，还原糖含量为0.5-1.0mg/mL，pH=6.0-6.5。
4.根据权利要求1所述的一种复合海藻多糖降血脂口服液，其特征在于，步骤（3）中，所述浓缩是浓缩至固形物含量为4~10%；加入食用酒精至酒精浓度为60~90 vol %；所述冷藏是在4℃下冷藏12小时；所述加水溶解是按照沉淀物与水的质量比1:10-30添加水。
5.根据权利要求1所述的一种复合海藻多糖降血脂口服液，其特征在于，所述甜味剂包括白砂糖、木糖醇、低聚果糖、低聚木糖、低聚异麦芽糖、大豆低聚糖、低聚半乳糖、蜂蜜、阿斯巴甜、三氯蔗糖和甜菊糖苷中的一种以上。
6.权利要求1~5任一项所述的一种复合海藻多糖降血脂口服液的制备方法，其特征在于，包括如下步骤：
按所述质量百分比将各组分进行混合，将混合得到的混合物在4℃冷藏保存12小时后，离心取上清液，灌装、封盖、灭菌，得到所述复合海藻多糖降血脂口服液。
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