CN111280449B - 一种肠内缓释糖醇添加剂及其制备方法和应用 - Google Patents

一种肠内缓释糖醇添加剂及其制备方法和应用 Download PDF

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CN111280449B
CN111280449B CN202010087312.6A CN202010087312A CN111280449B CN 111280449 B CN111280449 B CN 111280449B CN 202010087312 A CN202010087312 A CN 202010087312A CN 111280449 B CN111280449 B CN 111280449B
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sugar alcohol
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xylitol
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CN111280449A (zh
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程新平
李勉
朱炫
石丽华
叶昆
向沙沙
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Zhejiang Huakang Pharmaceutical Co Ltd
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Abstract

本发明涉及一种肠内缓释糖醇添加剂包括两层结构，在其内层结构中包含有木糖醇和琼脂的组分，在其外层结构中包含有卡拉胶或结冷胶或黄原胶或瓜尔豆胶、维生素B12、L‑阿拉伯糖以及发酵的双歧杆菌或发酵的丙酸杆菌或发酵的乳杆菌的组分。本发明还公开制备肠内缓释糖醇添加剂的方法及其应用。本发明使用复配的木糖醇、L‑阿拉伯糖和维生素B12，实现了肠道微生物定向调控，并实现肠内丙酸高浓度合成。

Description

一种肠内缓释糖醇添加剂及其制备方法和应用
技术领域
本发明属于肠内缓释产品的制备技术领域，特别涉及一种肠内缓释糖醇添加剂及其制备方法和应用。
背景技术
丙酸被吸收后主要在肝脏中被代谢，参与糖异生，抑制胆固醇合成，但已经有各种研究表明，外源性地摄入和自体产生的丙酸作用并不一样，外来的丙酸对个体的许多代谢过程有重要影响。丙酸（propionate）是一种短链脂肪酸，正常情况下，人体中的微生物是能够在结肠中通过发酵未完全消化的碳水化合物来产生丙酸的，这些丙酸对人体是有益的。而外源性丙酸在短期内会导致高血糖、胰岛素偏高，长期摄入将会造成肥胖和胰岛素抵抗等更严重的症状。
低聚果糖、多糖具有在肠道内生成丙酸的功能。但是无论低聚果糖还是多糖都不能定点、高浓度的合成丙酸。多糖的肠内利用率不高，且容易导致胀气等问题。低聚果糖同样会导致产气过多，与产酸不稳定等问题，甚至会产生果糖过多导致肝脏代谢紊乱问题。
发明内容
本发明所要解决的技术问题在于，提供一种肠内缓释糖醇添加剂及其制备方法和应用，促进机体健康同时，消除丙酸可能产生的代谢副作用，实现糖醇添加剂在肠道内的缓慢释放和结肠内定点释放。
本发明是这样实现的，提供一种肠内缓释糖醇添加剂，其包括两层结构，在其内层结构中包含有木糖醇和琼脂的组分，在其外层结构中包含有卡拉胶或结冷胶或黄原胶或瓜尔豆胶、维生素B12、L-阿拉伯糖以及发酵的双歧杆菌或发酵的丙酸杆菌或发酵的乳杆菌的组分。
本发明是这样实现的，还提供一种如前所述的肠内缓释糖醇添加剂的制备方法，包括以下步骤：
步骤一、将木糖醇与琼脂混合，灭菌，放入球型磨具静置冷却制成木糖醇琼脂小球；
步骤二、配制卡拉胶溶液或结冷胶溶液或黄原胶溶液或瓜尔豆胶溶液，将维生素B12、L-阿拉伯糖加入到卡拉胶溶液或结冷胶溶液或黄原胶溶液或瓜尔豆胶溶液中混匀得到混合液，再将该混合液包裹步骤一的木糖醇琼脂小球，得到包裹混合液的木糖醇琼脂的二层胶体小球；
步骤三、将步骤二制备的二层胶体小球放置于发酵的双歧杆菌培养液或发酵的丙酸杆菌培养液或发酵的乳杆菌培养液中培养，培养后取出再进行冷冻或热风干燥，得到糖醇添加剂成品。
本发明是这样实现的，还提供一种如前所述的肠内缓释糖醇添加剂的应用，所述糖醇添加剂应用于食品上。
本发明是这样实现的，还提供一种如前所述的肠内缓释糖醇添加剂的制备方法制备的肠内缓释糖醇添加剂的应用，所述糖醇添加剂应用于食品上。
与现有技术相比，本发明的肠内缓释糖醇添加剂及其制备方法和应用具有以下特点：
1. 使用木糖醇、L-阿拉伯糖和维生素B12，实现了肠道微生物定向调控，并实现肠内丙酸高浓度合成；
2. 使用分层包埋技术，实现了肠道内的糖醇缓释和分层释放；
3. 本方法的小球技术，实现了双歧杆菌类或丙酸杆菌类或乳杆菌类产品稳定性。
附图说明
图1为本发明的肠内缓释糖醇添加剂中的木糖醇组分在模拟胃、小肠和结肠消化过程中含量变化示意图；
图2为本发明的肠内缓释糖醇添加剂中的阿拉伯糖组分在模拟胃、小肠和结肠消化过程中含量变化示意图；
图3为本发明的二层胶体小球产品与肠内缓释糖醇添加剂成品在模拟结肠实验中双歧杆菌属的相对丰度对比示意图。
具体实施方式
为了使本发明所要解决的技术问题、技术方案及有益效果更加清楚明白，以下结合附图及实施例，对本发明进行进一步详细说明。应当理解，此处所描述的具体实施例仅仅用以解释本发明，并不用于限定本发明。
本发明的肠内缓释糖醇添加剂包括两层结构，在其内层结构中包含有木糖醇和琼脂的组分，在其外层结构中包含有卡拉胶或结冷胶或黄原胶或瓜尔豆胶、维生素B12、L-阿拉伯糖以及发酵的双歧杆菌或发酵的丙酸杆菌或发酵的乳杆菌的组分。
具体地，在所述糖醇添加剂中，所含的木糖醇、L-阿拉伯糖、维生素B12的组分比为1~33:1:0.005。
本发明的肠内缓释糖醇添加剂通过木糖醇、L-阿拉伯糖和维生素B12复配，L-阿拉伯糖减缓木酮糖向阿拉伯糖醇代谢并控制代谢流，维生素B12增加丙酸代谢途径，实现了首先诱导肠杆菌科和毛罗菌科菌与木糖醇相关的基因表达增加，从而增加木糖醇向PPPpathway途径代谢，减弱木酮糖代谢为阿拉伯糖醇途径，增加丙酸合成数量。
本发明的肠内缓释糖醇添加剂采用两层结构，以木糖醇为核心，用卡拉胶等食用胶制备L-阿拉伯糖和维生素B12的二层胶体，实现木糖醇和L-阿拉伯糖在肠道内的缓慢释放和结肠内定点释放。
本发明还公开一种如前所述的肠内缓释糖醇添加剂的制备方法，包括以下步骤：
步骤一、将木糖醇与琼脂混合，灭菌，放入球型磨具静置冷却制成木糖醇琼脂小球。
步骤二、配制卡拉胶溶液或结冷胶溶液或黄原胶溶液或瓜尔豆胶溶液，将维生素B12、L-阿拉伯糖加入到卡拉胶溶液或结冷胶溶液或黄原胶溶液或瓜尔豆胶溶液中混匀得到混合液，再将该混合液包裹步骤一的木糖醇琼脂小球，得到包裹混合液的木糖醇琼脂的二层胶体小球。
步骤三、将步骤二制备的二层胶体小球放置于发酵的双歧杆菌培养液或发酵的丙酸杆菌培养液或发酵的乳杆菌培养液中培养，培养后取出再进行冷冻或热风干燥，得到糖醇添加剂成品。
下面结合具体实施例进一步说明本发明的肠内缓释糖醇添加剂的制备方法。
实施例1
本发明的第一种肠内缓释糖醇添加剂的制备方法的实施例，包括以下步骤：
步骤一、称取一定量的木糖醇与2%的琼脂混合，121℃灭菌15min，待冷却至55℃时，用一次性无菌胶头滴管吸取转移到球型磨具中，静置冷却即可制成木糖醇琼脂小球。
步骤二、称取2g卡拉胶，溶解于100mL蒸馏水中，121℃灭菌15min，待冷却至50℃~60℃，温度不高于60℃，便于维生素B12稳定，避光，无菌L-阿拉伯糖、维生素B12按照混合配方比1:0.005加入到卡拉胶溶液中混匀，在无菌超净台中包裹到木糖醇琼脂小球制成配方比为1~33:1:0.005的二层胶体小球。
步骤三、将步骤二制备的二层胶体小球按照总体积的1：3添加入双歧杆菌发酵（109CFU/mL）液中，静止培养24小时；培养后取出带菌小球再进行冷冻干燥1h~24h或热风干燥30min，得到糖醇添加剂成品。
在上述实施例中，如果将双歧杆菌替换为丙酸杆菌或乳杆菌以得到新的实施例。
选取本实施例制备的肠内缓释糖醇添加剂进行胃消化、小肠消化和结肠消化模拟实验进行验证。实验方法如下：
1）模拟胃液准备：将0.62g的NaCl、0.22g的KCl、0.05g的CaCl2、0.12g的NaHCO3溶于200mL蒸馏水中，使用0.1M的HCl溶液调节pH至2.0，制得胃液培养基。然后，将1.0mL的CH3COONa(1.0mol/L，pH5)、23.6g胃蛋白酶和25.0g胃脂肪酶分别加入100mL胃培养基中，然后将培养基的pH值用0.1M的HCl溶液调整到2.0。
2）模拟胃液消化：按照木糖醇1%~5%的剂量，将实施例1的糖醇添加剂成品小球与模拟胃液体积1：1混合，37℃水浴0.5h、1h、2h、4h、6h，分别取出消化后的样品。
3）模拟小肠消化液制备：将0.54g的NaCl、0.065g的KCl、0.033g的CaCl2溶于100mL蒸馏水中，用0.1M的NaOH溶液调节pH至7.0，制备小肠培养基。在小肠培养基中分别加入6.5mg胰蛋白酶、200.0g胆盐(4%，w/w)和50.0g胰酶液(7%，w/w)。用0.1M的NaOH溶液，室温连续搅拌，调节溶液pH至7.5，模拟小肠消化液。
4）模拟小肠消化：将模拟胃液消化后的样品分别与小肠液体积比3：7混合，37℃水浴0.5h、1h、2h、4h、6h，分别取出消化后的样品。
5）将实施例1的二层胶体小球样品（无包埋双歧杆菌菌体）和糖醇添加剂成品小球样品（有包埋双歧杆菌菌体）按照木糖醇1%~5%剂量，补充到CDMN（Chang Dao Mo Ni）结肠体外模拟系统，收集模拟结肠消化24h、48h的小球样品。
6）使用HPLC分别测定模拟胃消化、模拟小肠消化、模拟结肠消化后保留在小球中的木糖醇、阿拉伯糖的含量，以及双歧杆菌属的丰度值，得到如图1至图3所示的对比曲线。
如图1和图2所示，以木糖醇：阿拉伯糖=25：1的小球为例，每个小球中木糖醇含量为1g，阿拉伯糖含量为0.04g。前6小时为胃消化、6~12小时为小肠消化、12~60小时为大肠微生物消化后小球中木糖醇和阿拉伯糖含量。说明本发明的小球对阿拉伯糖和木糖醇能够起缓慢、定点释放的效果。
如图3所示，本发明的小球内双歧杆菌包埋（糖醇添加剂成品）相比于不包埋（二层胶体小球产品），能够显著促进小球中双歧杆菌的相对丰度，从而促进结肠中双歧杆菌相对丰度。
因此，本发明的肠内缓释糖醇添加剂使用分层包埋技术，实现了肠道内的糖醇缓释和分层释放，而且，促进结肠中双歧杆菌的相对丰度。
本发明还公开一种如前所述的肠内缓释糖醇添加剂的应用，所述糖醇添加剂应用于食品上。
本发明还公开一种如前所述的肠内缓释糖醇添加剂的制备方法制备的肠内缓释糖醇添加剂的应用，所述糖醇添加剂应用于食品上。
例如本发明的肠内缓释糖醇添加剂被应用在饮料、糕点中，促进人体结肠内的丙酸的产生，调节肠道健康。

Claims (5)

1.一种肠内缓释糖醇添加剂的制备方法，其特征在于，肠内缓释糖醇添加剂包括两层结构，其内层结构为：木糖醇和琼脂；其外层结构为：卡拉胶或结冷胶或黄原胶或瓜尔豆胶中任意一种，维生素B12，L-阿拉伯糖以及发酵的双歧杆菌或发酵的丙酸杆菌或发酵的乳杆菌；在所述糖醇添加剂中，所含的木糖醇、L-阿拉伯糖、维生素B12的组分比为1~33:1:0.005，所述制备方法包括以下步骤：
步骤一、将木糖醇与琼脂混合，灭菌，放入球型模具静置冷却制成木糖醇琼脂小球，琼脂的浓度为2%；
步骤二、配制卡拉胶溶液或结冷胶溶液或黄原胶溶液或瓜尔豆胶溶液，将维生素B12、L-阿拉伯糖加入到卡拉胶溶液或结冷胶溶液或黄原胶溶液或瓜尔豆胶溶液中混匀得到混合液，再将该混合液包裹步骤一的木糖醇琼脂小球，得到包裹混合液的木糖醇琼脂的二层胶体小球；
步骤三、将步骤二制备的二层胶体小球放置于发酵的双歧杆菌培养液或发酵的丙酸杆菌培养液或发酵的乳杆菌培养液中培养，培养后取出再进行冷冻干燥或热风干燥，得到糖醇添加剂成品。
2.如权利要求1所述的肠内缓释糖醇添加剂的制备方法，其特征在于，在步骤一中，灭菌条件是121℃灭菌15min。
3.如权利要求1所述的肠内缓释糖醇添加剂的制备方法，其特征在于，在步骤二中，配制卡拉胶溶液或结冷胶溶液或黄原胶溶液或瓜尔豆胶溶液的方法包括：称取2g卡拉胶或结冷胶或黄原胶或瓜尔豆胶，溶解于100mL蒸馏水中，121℃灭菌15min，待冷却至温度50℃~60℃。
4.如权利要求1所述的肠内缓释糖醇添加剂的制备方法，其特征在于，在步骤三中，在发酵的双歧杆菌培养液或发酵的丙酸杆菌培养液或发酵的乳杆菌培养液中双歧杆菌或丙酸杆菌或乳杆菌浓度为109CFU/mL，二层胶体小球放置于发酵的双歧杆菌培养液或发酵的丙酸杆菌培养液或发酵的乳杆菌培养液中培养24h，冷冻干燥或热风干燥的条件是冷冻干燥1h~24h或热风干燥30min。
5.一种如权利要求1至4中任意一项所述的肠内缓释糖醇添加剂的制备方法制备的肠内缓释糖醇添加剂的应用，其特征在于，所述糖醇添加剂应用于食品上。
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