CN110651875A - 一种防控幽门螺杆菌的圈糖及其制备方法 - Google Patents
一种防控幽门螺杆菌的圈糖及其制备方法 Download PDFInfo
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Abstract
本发明公开了一种防控幽门螺杆菌的圈糖及其制备方法，具体涉及糖果技术领域，本发明制备的圈糖中包含有四价型的防控幽门螺杆菌的蛋黄免疫球蛋白。本发明制作出来的圈糖能够代替药物防控胃部及口腔幽门螺杆菌，降低胃病产生率，去除口腔异味，无副作用。
Description
技术领域
本发明属糖果技术领域，具体涉及一种防控幽门螺杆菌的圈糖及其制备方法。
背景技术
幽门螺杆菌或幽门螺旋菌，英文名Helicobacterpylori，简称Hp。是革兰氏阴性、微需氧的细菌，生存于胃部及十二指肠的各区域内。它会引起胃黏膜轻微的慢性发炎，甚或导致胃及十二指肠溃疡与胃癌。常用的治疗方法是采用三联疗法和四联疗法，常见的三联疗法是一种制酸药（奥美拉唑、雷贝拉唑等，都是质子泵制剂），加两种抗生素（一般是阿西莫林和克拉霉素，如果青霉素过敏者，可以使用甲硝唑加克拉霉素）；四联疗法就是在前三种的基础上加铋剂，如丽珠得乐、国胶铋等，这些药物不仅对人损害较大，而且对于一些不喜欢吃药的人来说，治疗起来比较困难。
因此，急需一种解决现有问题的防控幽门螺杆菌的圈糖及其制备方法。
发明内容
本发明的目的是提供一种防控幽门螺杆菌的圈糖及其制备方法，制作出来的圈糖能够代替药物防控胃部及口腔幽门螺杆菌，降低胃病产生率，去除口腔异味，无副作用。
技术方案：为了实现上述发明目的，本发明采用的技术方案为：
一种防控幽门螺杆菌的圈糖，包含有四价型的防控幽门螺杆菌的蛋黄免疫球蛋白。四价型的防控幽门螺杆菌的蛋黄免疫球蛋白的产品（四价指①HapA黏附素 ②Vac空泡毒素③CAG细胞毒素相关基因 ④URE尿素酶）通过抗幽门螺杆菌特异性IgY的高免蛋黄去影响Urel（Urel是一种专门负责调节胞质内尿素酶的蛋白质，起“质子通道”作用，可控制进入菌体的胃酸量），使菌体失去保护，无法在强酸环境下存活，从而有效抑制或消灭幽门螺杆菌；通过圈糖的形式直接吃进胃中，代替药物，直接抑制或消灭幽门螺杆菌，特别适合不喜欢吃药的人群。
优选的，各原料质量配比如下：
山梨糖醇30-60份、DL-苹果酸0.5-1份、海盐0.2-2份、三氯庶糖30-60份、硬脂酸镁0.2-3份、维生素C0.1-0.5份、食用香精0.2-5份、粘合剂3-5份和蛋黄免疫球蛋白0.05-0.3份。
优选的，各原料质量配比如下：
山梨糖醇46份、DL-苹果酸0. 7份、海盐0.9份、三氯庶糖45份、硬脂酸镁1.2份、维生素C0.3份、食用香精1.8份、粘合剂4份和蛋黄免疫球蛋白0.1份。
一种权利要求2或3所述的防控幽门螺杆菌的圈糖的制备方法，其特征在于，包括如下步骤：
S1、初混合：按照对应的配比将山梨糖醇、DL-苹果酸、海盐、三氯庶糖、维生素C进行混合；
S2、制粒：将步骤S1中初混合过程完成的物料用粘合剂制成湿颗粒；
S3、烘干；
S4、整理；
S5、总混合：将整粒过程完成后的物料，加入对应质量的蛋黄免疫球蛋白、食用香精、硬脂酸镁进行总混合；
S6、压片；
S7、拣片。
优选的，初混合和总混合使用的混合设备为三维摆动混合机。
一种蛋黄免疫球蛋白的制备方法如下：
1）以幽门螺杆菌菌株作为抗原，与弗氏不完全佐剂1:1混合并吹打使其乳化均匀得到幽门螺杆菌菌株乳化液；
2）在蛋鸡的两侧鸡翅及大腿根部注射步骤1）获得的幽门螺杆菌菌株乳化液，每个部位0.25 mL，每7天免疫一次，共免疫4次；
3）免疫后每日收集并标记鸡蛋，提取蛋黄液中的蛋黄免疫球蛋白，冷冻干燥成产品。
有益效果：
1、防控胃部及口腔幽门螺杆菌，降低胃病产生率，去除口腔异味；
2、通过圈糖的形式直接吃进胃中，口感好，特别适合不喜欢吃药的人群，无副作用。
具体实施方式
实施例1
四价型的防控幽门螺杆菌的蛋黄免疫球蛋白的制备：
（1）蛋鸡免疫
38周龄的白来航蛋鸡，饲养于超洁净的工作台中，饲料与水(pH 6.2需要经过高压蒸汽灭菌。蛋鸡免疫前，以菌体浓度为109-1011CFU/mL的幽门螺杆菌菌株作为抗原，与弗氏不完全佐剂1:1混合并吹打使其乳化均匀获得幽门螺杆菌菌株乳化液。在实验组（A组）蛋鸡的两侧鸡翅及大腿根部注射预先处理好的幽门螺杆菌菌株乳化液，每个部位0.25 mL，每7天免疫一次，共免疫4次。对照组（B组）蛋鸡注射生理盐水和弗氏不完全佐剂的等体积混合液。给药部位和剂量同实验组。免疫后每日收集并标记鸡蛋，4℃存储。
（2）提取和纯化免疫球蛋白（IgY）
用医用酒精棉擦拭鸡蛋表面，磕破鸡蛋外壳，用10 mL注射器抽取蛋黄液并记录体积，用预冷的超纯水(1:9)稀释并充分混合(0.1 mol HCl调节至pH 5.0)，然后在4℃下储存过夜。将溶液在5000 g, 4℃下离心45 min,向上清液中加入硫酸铵使最终饱和度为45，然后在4℃下放置2.5 h。在5000 g, 4℃下离心12 min后，弃去上清液，1:1加超纯水溶解蛋白质沉淀，然后加入硫酸钠使溶液的最终质量分数达到13，室温搅拌溶解后静置2.5h; 4℃条件下5000 g再次离心12 min，并将沉淀物溶解PBS中。将溶液透析6h(每1.5 h更换一次超纯水)，然后在PBS中浸泡过夜并在一20℃下储存。免疫后从A组卵中纯化的IgY是特异性IgY（S-IgY ) , B组是阴性对照IgY （C-IgY）。用Amicon Ultra-15离心过滤装置(Millipore，伯灵顿，美国)将纯化的IgY以4000 g超滤离心10 min以达到脱盐和浓缩抗体的目的，冷冻干燥成产品。
以B组作对照，采用ELISA(酶联免疫吸附实验)测得A组的特异性卵黄抗体效价达1:2450000。
四价型的防控幽门螺杆菌的蛋黄免疫球蛋白（四价指①HapA黏附素 ②Vac空泡毒素 ③CAG细胞毒素相关基因 ④URE尿素酶）通过抗幽门螺杆菌特异性IgY的高免蛋黄去影响Urel（Urel是一种专门负责调节胞质内尿素酶的蛋白质，起“质子通道”作用，可控制进入菌体的胃酸量），使菌体失去保护，无法在强酸环境下存活，从而有效抑制或消灭幽门螺杆菌。
实施例2
按照如下配比制作对照组圈糖和3组实验组圈糖，三组实验组圈糖分别命名为实验1、实验2和实验3。
表1 圈糖原料配比
| 实验1 | 实验2 | 实验3 | 对照组 | |
| 山梨糖醇（份） | 46 | 31 | 58 | 46 |
| DL-苹果酸（份） | 0.7 | 1 | 0.5 | 0.7 |
| 海盐（份） | 0.9 | 1.6 | 0.4 | 0.9 |
| 三氯庶糖（份） | 45 | 60 | 31 | 45 |
| 硬脂酸镁（份） | 1.2 | 0.45 | 2.3 | 1.2 |
| 维生素C（份） | 0.3 | 0.1 | 0.5 | 0.4 |
| 食用香精（份） | 1.8 | 0.8 | 4 | 1.8 |
| 粘合剂（份） | 4 | 5 | 3 | 4 |
| 蛋黄免疫球蛋白（份） | 0.1 | 0.05 | 0.3 | 0 |
本发明中的粘合剂可以为蔗糖溶液、淀粉浆中的任意一种，在本实施例中选用蔗糖溶液。
圈糖的制备方法如下：
配料：根据配方量取所需要的山梨糖醇、DL-苹果酸、海盐、三氯庶糖、硬脂酸镁、维生素C、食用香精、粘合剂和蛋黄免疫球蛋白。
初混合：将山梨糖醇、DL-苹果酸、海盐、三氯庶糖、维生素C放入三维摆动混合机进行三维摆动混合的操作过程。
制粒：将初混合过程完成的物料用粘合剂制成湿颗粒的操作过程。使用快速搅拌制粒机进行快速搅拌。
烘干：将制粒过程完成后的物料，进行烘干的操作过程。使用热风循环烘箱设备进行烘干。
整粒：将烘干过程完成后的物料，进行整粒的操作过程。使用高速粉碎整粒机设备。
总混合：将整粒过程完成后的物料，加入IgY（蛋黄免疫球蛋白）、食用香精、硬脂酸镁进行总混合的操作过程。使用三维摆动混合机进行混合。质保部QA质监员对此必须进行监控，并对每锅混合物料进行水分检测（中间产品检验属于关键质量控制点），并做好相关控制记录。
压片：将混合过程完成后的物料进行压片的操作过程。使用旋转式压片机设备进行压片。
压片工序属于关键质量控制点，质保部QA质监员对此必须进行控制，在压片过程中要定时进行片重的抽检复核，并做好相关控制记录。
拣片：对压片过程完成后的素片挑拣的操作过程。
本方法制作的压片糖果也就是圈糖的质量为1000mg。
实施例3
性能测试
考察本产品防控幽门螺杆菌及去口腔异味实验方法：选择240例Hp阳性患者，分为治疗组1（50例）、治疗组2（50例）、治疗组3( 50例)和对照组( 50例)，治疗组1、治疗组2以及治疗组3分别服用实施例2中的实验1、实验2以及实验3获得的圈糖1粒，每天2次(早晚各一次)，对照组服用实施例2中的对照组获得的圈糖1粒，每天早晚2次，使用周期为1个月，停药1个月后统计变阴率及口腔异味去除情况，结果如下：
表2：防控幽门螺杆菌实验结果
| 治疗组1 | 治疗组2 | 治疗组3 | 对照组 | |
| 转阴（人） | 24 | 17 | 25 | 0 |
| 口腔异味去除（人） | 30 | 25 | 33 | 0 |
| 口腔异味得到明显改善（人） | 50 | 50 | 50 | 0 |
以上实验数据证明本产品效果显著，四价型的防控幽门螺杆菌的蛋黄免疫球蛋白在其中起到了显著的作用，随着其浓度增加，效果随之变好，但达到一定浓度后，增加浓度，效果增加变化不明显。
值得注意的是口腔异味得到明显改善包含口腔异味去除。
以上所述，仅为本发明较佳的具体实施方式，但本发明的保护范围并不局限于此，任何熟悉本技术领域的技术人员在本发明揭露的技术范围内，根据本发明的技术方案及其发明构思加以等同替换或改变，都应涵盖在本发明的保护范围之内。
Claims (6)
1.一种防控幽门螺杆菌的圈糖，其特征在于，包含有四价型的防控幽门螺杆菌的蛋黄免疫球蛋白。
2.根据权利要求1所述的防控幽门螺杆菌的圈糖，其特征在于，各原料质量配比如下：
山梨糖醇30-60份、DL-苹果酸0.5-1份、海盐0.2-2份、三氯庶糖30-60份、硬脂酸镁0.2-3份、维生素C0.1-0.5份、食用香精0.2-5份、粘合剂3-5份和蛋黄免疫球蛋白0.05-0.3份。
3.根据权利要求2所述的防控幽门螺杆菌的圈糖，其特征在于，各原料质量配比如下：
山梨糖醇46份、DL-苹果酸0. 7份、海盐0.9份、三氯庶糖45份、硬脂酸镁1.2份、维生素C0.3份、食用香精1.8份、粘合剂4份和蛋黄免疫球蛋白0.1份。
4.一种权利要求2或3所述的防控幽门螺杆菌的圈糖的制备方法，其特征在于，包括如下步骤：
S1、初混合：按照对应的配比将山梨糖醇、DL-苹果酸、海盐、三氯庶糖、维生素C进行混合；
S2、制粒：将步骤S1中初混合过程完成的物料用粘合剂制成湿颗粒；
S3、烘干；
S4、整理；
S5、总混合：将整粒过程完成后的物料，加入对应质量的蛋黄免疫球蛋白、食用香精、硬脂酸镁进行总混合；
S6、压片；
S7、拣片。
5.根据权利要求4所述的防控幽门螺杆菌的圈糖的制备方法，其特征在于，步骤S5中的蛋黄免疫球蛋白的制备方法如下：
1）以幽门螺杆菌菌株作为抗原，与弗氏不完全佐剂1:1混合并吹打使其乳化均匀得到幽门螺杆菌菌株乳化液；
2）在蛋鸡的两侧鸡翅及大腿根部注射步骤1）获得的幽门螺杆菌菌株乳化液，每个部位0.25 mL，每7天免疫一次，共免疫4次；
3）免疫后每日收集并标记鸡蛋，提取蛋黄液中的蛋黄免疫球蛋白，冷冻干燥成产品。
6.根据权利要求4所述的防控幽门螺杆菌的圈糖的制备方法，其特征在于，初混合和总混合使用的混合设备为三维摆动混合机。
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