海藻酸钙微胶囊包埋IgY及其释放特性研究

doi:10.3969/j.issn.1004-1524.2022.12.17

浙江农业学报 ›› 2022, Vol. 34 ›› Issue (12): 2740-2749.DOI: 10.3969/j.issn.1004-1524.2022.12.17

海藻酸钙微胶囊包埋IgY及其释放特性研究

卢文静¹(), 谌迪¹, 叶沁¹, 张岑¹, 阮玲丽¹^,², 肖朝耿¹^,^*(), 姚云昕³

1.浙江省农业科学院食品科学研究所,浙江杭州 310021
2.云南农业大学食品科学技术学院,云南昆明 650201
3.浙江艾格生物科技股份有限公司,浙江长兴 313100

收稿日期:2022-03-28 出版日期:2022-12-25 发布日期:2022-12-26
作者简介:*肖朝耿,E-mail: xiaochaogeng@163.com
卢文静(1987—),女,山东蒙阴人,博士,助理研究员,研究方向为功能性食品配料。E-mail:wenjing_316@163.com
通讯作者: 肖朝耿
基金资助:
浙江省基础公益研究计划(LGN22C200043);“科技助力经济2020”国家重点专项(SQ2020YFF0421795)

Study on calcium alginate microcapsule embedding IgY and its release properties

LU Wenjing¹(), CHEN Di¹, YE Qin¹, ZHANG Cen¹, RUAN Lingli¹^,², XIAO Chaogeng¹^,^*(), YAO Yunxin³

1. Institute of Food Science, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China
2. College of Food Science and Technology, Yunnan Agricultural University, Kunming 650201, China
3. Zhejiang AGS Biotechnology Company Limited, Changxing 313100, Zhejiang, China

Received:2022-03-28 Online:2022-12-25 Published:2022-12-26
Contact: XIAO Chaogeng

摘要/Abstract

摘要：

卵黄免疫球蛋白(IgY)是一种高活性的口服抗体,与人体健康密切相关。为最大限度保留IgY的生物活性,提高其稳定性,以海藻酸钠为主要壁材,使用锐孔-凝固浴法包埋IgY制备凝胶珠。以包埋率和载药量为主要指标,利用响应面法探究IgY-海藻酸钙凝胶珠的最佳制备工艺,并利用体外胃肠道消化模型评估凝胶珠释放效果。结果显示:当海藻酸钠为28.30 g·L^-1,氯化钙为10.80 g·L^-1,芯壁比(质量比)为0.46∶1时,新鲜制备得到的IgY-海藻酸钙凝胶珠成型效果较好,包埋率为(53.30±1.55)%,载药量为(10.90±0.26)%。通过宏观形态观察,凝胶珠颗粒具有较好的圆整性和均一性,粒径约为(2.10±0.05) mm,硬度为(4 396.87±331.62) g。干燥后的凝胶珠在模拟胃液中2 h后溶胀度为0.94±0.39,IgY累积释放率约在(11.10±3.20)%,在模拟肠液中2 h后,溶胀度持续增加至4.81±0.32,IgY累积释放率达(97.10±2.40)%以上。因此,IgY-海藻酸钙凝胶珠在胃液中具有缓慢释放芯材的作用,IgY在肠液中能基本全部释放。

关键词: 鸡卵黄免疫球蛋白, 海藻酸钙, 海藻酸钠, 体外释放率

Abstract:

Egg yolk immunoglobulin (IgY) is a highly active oral antibody that is closely related to human health. In order to maximize the retention of IgY's biological activity and improve its stability, the natural polysaccharide sodium alginate was used as the main wall material, and the gel bead was prepared by the method of retination-coagulation bath embedding IgY. Taking the encapsulation efficiency and loading capacity as the main indicators, the optimal preparation process of IgY-calcium sodium alginate gel beads was explored by the response surface method, and the in vitro gastrointestinal digestion model was used to evaluate the release effect of gel beads. The results showed that when sodium alginate was 28.30 g·L^-1, calcium chloride was 10.80 g·L^-1 and the mass ratio of core to wall was 0.46:1, the newly prepared IgY-calcium alginate gel beads had good molding effect, the encapsulation efficiency was (53.30±1.55)%, and the loading capacity was (10.90±0.26)%. Through macroscopic morphological observation, the gel bead particles had good roundness and uniformity, with a particle size of about (2.10±0.05) mm and a hardness of (4 396.87±331.62) g. After drying, the swelling ratio of the gel beads was 0.94±0.39 after 2 h in the simulated gastric juice, the cumulative release ratio of IgY was about (11.10±3.20)%, and after 2 h in the simulated intestinal fluid, the swelling ratio continued to increase to 4.81±0.32, and the cumulative release ratio of IgY reached (97.10±2.40)%. Therefore, IgY-calcium alginate gel beads had the effect of slowly releasing the core material in the gastric juice, and IgY could be basically released in the intestinal fluid.

Key words: egg yolk immunoglobulins, calcium alginate, sodium alginate, external release rate

中图分类号:

TQ460.4

卢文静, 谌迪, 叶沁, 张岑, 阮玲丽, 肖朝耿, 姚云昕. 海藻酸钙微胶囊包埋IgY及其释放特性研究[J]. 浙江农业学报, 2022, 34(12): 2740-2749.

LU Wenjing, CHEN Di, YE Qin, ZHANG Cen, RUAN Lingli, XIAO Chaogeng, YAO Yunxin. Study on calcium alginate microcapsule embedding IgY and its release properties[J]. Acta Agriculturae Zhejiangensis, 2022, 34(12): 2740-2749.

图/表 10

表1 凝胶珠因素水平表

Table 1 Horizontal table of gel bead factors

水平 Level	海藻酸钠 Sodium alginate/ (g·L^-1)	氯化钙 Calcium chloride/ (g·L^-1)	芯壁比 Core-to-wall ratio
1	30	20	4∶6
0	25	15	3∶6
-1	20	10	2∶6

图1 海藻酸钠质量浓度对凝胶珠包埋效果的影响图中同一标签标示不同小写字母表示差异显著(P<0.05)。

Fig.1 Influence of sodium alginate concentration on gel bead embedding effect Different lowercase letters in the figure indicated significant difference (P<0.05).

图2 CaCl2对凝胶珠包埋效果的影响

Fig.2 Influence of CaCl2 concentration on gel bead embedding effect

图3 芯壁比对凝胶珠包埋效果的影响

Fig.3 Influence of core-wall ratio on gel bead embedding effect

表2 凝胶珠响应面试验设计与结果

Table 2 Response surface test design and results of gel bead

试验号 Number	壁材浓度(X₁) Wall material concentration (X₁)/(g·L^-1)	氯化钙浓度(X₂) Calcium chloride concentration (X₂)/(g·L^-1)	芯壁比(X₃) Core-to-wall ratio (X₃)	包埋率 Encapsulation efficiency/%	载药量 Loading capacity/%
1	20	10	3∶6	29.55	8.79
2	30	15	4∶6	32.85	11.73
3	25	15	3∶6	51.37	15.29
4	20	20	3∶6	19.34	5.76
5	25	15	3∶6	50.91	15.15
6	30	15	2∶6	42.68	9.53
7	25	15	3∶6	53.87	16.03
8	25	15	3∶6	53.14	15.82
9	20	15	2∶6	7.55	1.67
10	30	20	3∶6	40.19	11.96
11	20	15	4∶6	33.53	11.79
12	25	10	4∶6	43.27	15.42
13	25	20	4∶6	49.85	17.76
14	25	10	2∶6	39.72	8.84
15	30	10	3∶6	54.27	16.15
16	25	20	2∶6	23.18	5.16
17	25	15	3∶6	37.63	11.20

表3 凝胶珠包埋率回归模型方差分析

Table 3 Variance analysis of regression model for gel bead embedding rate

方差来源 Source of variance	平方和 Sum of squares	自由度 Degree of freedom	均方 Mean square	F值 F value	P值 P value
模型Model	2 641.37	9	293.49	8.40	0.005 2
X₁	800.48	1	800.48	22.91	0.002 0
X₂	146.73	1	146.73	4.20	0.079 6
X₃	268.75	1	268.75	7.69	0.027 6
X₁X₂	3.78	1	3.78	0.11	0.751 9
X₁X₃	320.73	1	320.73	9.18	0.019 1
X₂X₃	133.60	1	133.60	3.82	0.091 4
$X 12$	576.69	1	576.69	16.51	0.004 8
$X 22$	14.37	1	14.37	0.41	0.541 8
$X 32$	306.42	1	306.42	8.77	0.021 1
残差Residual	244.54	7	34.93	—	—
失拟项Missing item	65.85	3	21.95	0.49	0.707 1
误差项Error term	178.70	4	44.67	—	—
总和Sum	2 885.92	16	—	—	—

表3 凝胶珠包埋率回归模型方差分析

Table 3 Variance analysis of regression model for gel bead embedding rate

方差来源 Source of variance	平方和 Sum of squares	自由度 Degree of freedom	均方 Mean square	F值 F value	P值 P value
模型Model	2 641.37	9	293.49	8.40	0.005 2
X₁	800.48	1	800.48	22.91	0.002 0
X₂	146.73	1	146.73	4.20	0.079 6
X₃	268.75	1	268.75	7.69	0.027 6
X₁X₂	3.78	1	3.78	0.11	0.751 9
X₁X₃	320.73	1	320.73	9.18	0.019 1
X₂X₃	133.60	1	133.60	3.82	0.091 4
$X 12$	576.69	1	576.69	16.51	0.004 8
$X 22$	14.37	1	14.37	0.41	0.541 8
$X 32$	306.42	1	306.42	8.77	0.021 1
残差Residual	244.54	7	34.93	—	—
失拟项Missing item	65.85	3	21.95	0.49	0.707 1
误差项Error term	178.70	4	44.67	—	—
总和Sum	2 885.92	16	—	—	—

表4 凝胶珠载药量回归模型方差分析

Table 4 Variance analysis of regression model of gel bead load

方差来源 Source of variance	平方和 Sum of squares	自由度 Degree of freedom	均方 Mean square	F值 F value	P值 P value
模型Model	301.00	9	33.44	7.76	0.006 6
X₁	57.03	1	57.03	13.23	0.008 3
X₂	9.16	1	9.16	2.12	0.188 3
X₃	124.03	1	124.03	28.77	0.001 0
X₁X₂	0.34	1	0.34	0.078	0.788 0
X₁X₃	15.68	1	15.68	3.64	0.098 1
X₂X₃	9.06	1	9.06	2.10	0.190 4
$X 12$	53.78	1	53.78	12.48	0.009 6
$X 22$	0.89	1	0.89	0.21	0.663 8
$X 32$	25.15	1	25.15	5.83	0.046 4
残差Residual	30.18	7	4.31	—	—
失拟项Missing item	14.35	3	4.78	1.21	0.414 0
误差项Error term	15.82	4	3.96	—	—
总和Sum	331.18	16	—	—	—

表4 凝胶珠载药量回归模型方差分析

Table 4 Variance analysis of regression model of gel bead load

方差来源 Source of variance	平方和 Sum of squares	自由度 Degree of freedom	均方 Mean square	F值 F value	P值 P value
模型Model	301.00	9	33.44	7.76	0.006 6
X₁	57.03	1	57.03	13.23	0.008 3
X₂	9.16	1	9.16	2.12	0.188 3
X₃	124.03	1	124.03	28.77	0.001 0
X₁X₂	0.34	1	0.34	0.078	0.788 0
X₁X₃	15.68	1	15.68	3.64	0.098 1
X₂X₃	9.06	1	9.06	2.10	0.190 4
$X 12$	53.78	1	53.78	12.48	0.009 6
$X 22$	0.89	1	0.89	0.21	0.663 8
$X 32$	25.15	1	25.15	5.83	0.046 4
残差Residual	30.18	7	4.31	—	—
失拟项Missing item	14.35	3	4.78	1.21	0.414 0
误差项Error term	15.82	4	3.96	—	—
总和Sum	331.18	16	—	—	—

图4 各因素交互作用对凝胶珠包埋率和载药量影响的响应面图

Fig.4 Response surface diagram of the interaction of the various factors on the encapsulation efficiency and loading capacity of gel bead

图5 IgY-海藻酸钙凝胶珠的宏观形态(×50倍)

Fig.5 Macro morphology of IgY-calcium alginate gel beads (×50 times)

表5 IgY-海藻酸钙凝胶珠在胃肠消化阶段的溶胀度与体外释放率

Table 5 Swelling ratio and release ratio of IgY-calcium alginate gel beads during the gastrointestinal digestion phase

消化时间 Digestion time/h		溶胀度 Swelling ratio	释放率 Release ratio/%
胃Stomach	1	0.90±0.24	9.80±2.40
	2	0.94±0.39	11.10±3.20
肠Intestine	1	2.26±0.15	91.40±4.20
	2	4.81±0.32	97.10±2.40

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海藻酸钙微胶囊包埋IgY及其释放特性研究

Study on calcium alginate microcapsule embedding IgY and its release properties

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