CN110511399A - 一种控释型纳米纤维素抗菌微凝胶的制备方法 - Google Patents

一种控释型纳米纤维素抗菌微凝胶的制备方法 Download PDF

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CN110511399A
CN110511399A CN201910680386.8A CN201910680386A CN110511399A CN 110511399 A CN110511399 A CN 110511399A CN 201910680386 A CN201910680386 A CN 201910680386A CN 110511399 A CN110511399 A CN 110511399A
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鲁鹏
葛小会
吴敏
郑璐
王志伟
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Abstract

本发明公开了一种控释型纳米纤维素抗菌微凝胶的制备方法,该方法包括如下步骤:(1)制备TEMPO氧化纳米纤维素胶体;(2)制备乳酸链球菌素溶液;(3)将TEMPO氧化纳米纤维素胶体置于聚四氟乙烯模具中,将乳酸链球菌素溶液滴加到TEMPO氧化纳米纤维素胶体表层形成水凝胶,再用去离子水浸泡冲洗得到纳米纤维素抗菌水凝胶;(4)将去离子水加入到水凝胶中,经机械分散法处理5min,得到控释型纳米纤维素抗菌微凝胶。本发明方法工艺简单,原料环境友好,制备的纳米纤维素微凝胶具有良好的控释抗菌性能,在农业、医学材料、组织工程等方面具有广阔的应用前景。

Description

一种控释型纳米纤维素抗菌微凝胶的制备方法
技术领域
本发明属于功能微凝胶制备及抗菌材料领域,特别是涉及一种控释型纳米纤维素抗菌微凝胶的制备方法。
背景技术
微凝胶是指尺寸在微纳米级别的凝胶,与宏观凝胶类似,是具有一定凝胶状三维立体网络结构的聚合物微粒,具有良好的生物相容性、多孔性和优异的加工性能,且在载药与可控释放、催化分离、化学传感器等多个领域具有广泛的应用。目前,微凝胶可以通过单体合成、高分子合成或通过大分子凝胶研磨获得。将微凝胶与抗菌剂相结合,可以获得抗菌性能优良的智能微凝胶,进一步的,通过调节环境pH、温度等因素控制抗菌剂的释放,这种控释型抗菌微凝胶在食品、生物医药等领域应用前景广泛。
乳酸链球菌素是由乳酸链球菌分泌的一种天然生物活性抗菌肽,它可以有效抑制或杀死食品腐败菌和致病菌在内等众多革兰氏阳性菌,是FDA公认的一种安全无毒的食品添加剂和防腐剂。将乳酸链球菌素与天然高分子材料进行结合,制备出乳酸链球菌素可控释放的功能材料,最大效率地发挥出抗菌效果,是其目前抗菌材料领域发展的主要趋势之一。纳米纤维素是尺寸在纳米范围内的纤维素,其来源广泛、成本较低,具有良好的相容性和生物可降解性。以纳米纤维素为原料,通过溶剂溶解置换、化学改性交联等方法可以获得结构和功能各异的水凝胶,近几年来作为制备新型水凝胶的材料而逐渐成为研究热点。
目前,有关乳酸链球菌素和纳米纤维素复合抗菌材料的报道很多,但是乳酸链球菌素的控释大多是通过直接混合包埋或是外加化学交联剂交联的方式来实现。目前,采用乳酸链球菌素负载纳米纤维素水凝胶的报道很少,直接利用乳酸链球菌素作为抗菌剂和交联剂制备纳米纤维素水凝胶的报道尚未见有报道。
发明内容
本发明所要解决的技术问题是提供一种控释型纳米纤维素抗菌微凝胶的制备方法,该方法工艺简单,不需加入催化剂、引发剂,原料来源绿色环保,得到的产品抗菌性能优异且抗菌剂可控释,具有很高的应用价值。
本发明以如下技术方案解决上述技术问题:
本发明一种控释型纳米纤维素抗菌微凝胶的制备方法,包括如下操作步骤:
(1)制备TEMPO氧化纳米纤维素胶体:采用TEMPO氧化法对纳米纤维素进行预处理,以得到表面羧基含量为600~1200mmol/Kg的TEMPO氧化纳米纤维素,然后采用溶剂蒸发法浓缩至质量浓度为3%,真空脱气,得到TEMPO氧化纳米纤维素胶体;
(2)制备乳酸链球菌素溶液:按重量份配比为2:98的比例将乳酸链球菌素粉末溶解在浓度为0.01M的盐酸水溶液中,得到质量浓度为2%的乳酸链球菌素溶液;
(3)凝胶化过程:将步骤(1)制得的TEMPO氧化纳米纤维素胶体,置于聚四氟乙烯模具中,再将步骤(2)制得的乳酸链球菌素溶液,用蠕动注射泵以1mL/min的速度沿模具壁滴加到TEMPO氧化纳米纤维素胶体表层,TEMPO氧化纳米纤维素胶体与乳酸链球菌素溶液的用量按重量份配比为40~80:20~60,于4℃下静置24~48h后形成水凝胶,然后将该水凝胶用去离子水浸泡冲洗2次,每次1分钟,得到纳米纤维素抗菌水凝胶;
(4)微凝胶:将去离子水加入到步骤(3)制得的水凝胶中,去离子水与水凝胶的用量按重量份配比为60~90:10~40,然后采用机械分散法处理5min,得到控释型纳米纤维素抗菌微凝胶。
本发明所述步骤(1)中的纳米纤维素是指将漂白针叶木纤维、漂白阔叶木纤维或漂白蔗渣纤维采用机械研磨法制备得到直径2~20nm、长度0.1~100μm的纳米纤维素。
本发明所述步骤(4)中的机械分散法是指采用超声细胞粉碎机、高速分散机或高压均质机进行处理。
本发明采用的乳酸链球菌素可以在酸性溶液中溶解并发生质子化,从而使分子表面带正电。以带阳离子电荷的乳酸链球菌素分子为交联点,运用其与带相反电荷的TEMPO氧化纳米纤维素之间的静电作用形成水凝胶,然后采用机械法将该水凝胶研磨即可获得微凝胶。由于乳酸链球菌素的等电点为pH=8.5,因此,通过调节微凝胶分散体系环境的pH值,即可控制微凝胶中负载的抗菌剂乳酸链球菌素的释放过程,从而获得控释型纳米纤维素抗菌水凝胶。
与现有技术相比,本发明具有如下有益效果:
1、本发明利用乳酸链球菌素与TEMPO氧化纳米纤维素胶体之间的静电吸附偶联作用,直接一步得到纳米纤维素水凝胶,然后借助机械分散法获得微凝胶。凝胶过程无需采用自由基聚合等方法,免去了外加单体、引发剂等的使用,制备工艺简单。
2、本发明所制备的微凝胶粒径均匀,呈单分散正态分布,在水中的稳定性和分散性能良好;随着食品体系的pH值因微生物生长繁殖导致的变化,即微凝胶分散体系所处环境的pH值变化,从而可控制微凝胶中负载的抗菌剂乳酸链球菌素的释放过程,在智能抗菌、食品、生物医药等领域有潜在的应用价值。
3、本发明采用的原材料纳米纤维素和乳酸链球菌素均为可再生生物质资源,属绿色环保材料,能够有效解决生产中的资源与环境压力。
具体实施方式
下面结合具体实施例对本发明的技术方案作进一步描述,但本发明不局限于以下实施例,以下实施例中所使用的化学试剂和原料均为分析纯。
实施例1
一种控释型纳米纤维素抗菌微凝胶的制备方法,具体操作步骤如下:
(1)制备TEMPO氧化纳米纤维素胶体:采用TEMPO氧化法对漂白针叶木纤维经机械研磨得到的直径2~20nm、长度0.1~100μm的纳米纤维素进行预处理,得到表面羧基含量为600mmol/Kg的TEMPO氧化纳米纤维素,然后采用溶剂蒸发法浓缩至质量浓度为3%,真空脱气,得到TEMPO氧化纳米纤维素胶体;
(2)制备乳酸链球菌素溶液:将乳酸链球菌素粉末溶解在浓度为0.01M的盐酸水溶液中,乳酸链球菌素与盐酸水溶液的用量按重量份配比为2:98,溶解后得到质量浓度为2%的乳酸链球菌素溶液;
(3)凝胶化过程:将步骤(1)制得的TEMPO氧化纳米纤维素胶体,置于聚四氟乙烯模具中,再将步骤(2)制得的乳酸链球菌素溶液,用蠕动注射泵以1mL/min的速度沿模具壁滴加到TEMPO氧化纳米纤维素胶体表层,TEMPO氧化纳米纤维素胶体与乳酸链球菌素溶液的用量按重量份配比为40:60,再于4℃下静置48h后形成水凝胶,然后将该水凝胶用等体积的去离子水浸泡冲洗2次,每次1分钟,得到纳米纤维素抗菌水凝胶;
(4)微凝胶:将去离子水加入到步骤(3)所制得的水凝胶中,去离子水与水凝胶的用量按重量份配比为80:20,再采用超声细胞粉碎机于1000W功率下处理5min,得到控释型纳米纤维素抗菌微凝胶。
通过激光粒度分布仪测得微凝胶的平均粒径范围为10~20μm,放置3天后无明显分层现象。将所得控释型纳米纤维素抗菌微凝胶胶涂覆在滤纸上干燥后进行抑菌圈测试,以李斯特菌为实验菌种测得第2天抑菌圈为8.7mm,第4天抑菌圈为11.2mm,第6天抑菌圈为12.6mm,表明控释型纳米纤维素抗菌微凝胶能有效控制抗菌剂乳酸链球菌素的释放速度,具有良好的控释抗菌性能。
实施例2
一种控释型纳米纤维素抗菌微凝胶的制备方法,具体操作步骤如下:
(1)制备TEMPO氧化纳米纤维素胶体:采用TEMPO氧化法对漂白蔗渣纤维经机械研磨得到的直径2~20nm、长度0.1~100μm的纳米纤维素进行预处理,得到表面羧基含量为900mmol/Kg的TEMPO氧化纳米纤维素,然后采用溶剂蒸发法浓缩至质量浓度为3%,真空脱气,得到TEMPO氧化纳米纤维素胶体;
(2)制备乳酸链球菌素溶液:将乳酸链球菌素粉末溶解在浓度为0.01M的盐酸水溶液中,乳酸链球菌素与盐酸水溶液的用量按重量份配比为2:98,溶解后得到质量浓度为2%的乳酸链球菌素溶液;
(3)凝胶化过程:将步骤(1)制得的TEMPO氧化纳米纤维素胶体,置于聚四氟乙烯模具中,再将步骤(2)制得的乳酸链球菌素溶液,用蠕动注射泵以1mL/min的速度沿模具壁滴加到TEMPO氧化纳米纤维素胶体表层,TEMPO氧化纳米纤维素胶体与乳酸链球菌素溶液的用量按重量份配比为60:40,再于4℃下静置36h后形成水凝胶,然后将该水凝胶用等体积的去离子水浸泡冲洗2次,每次1分钟,得到纳米纤维素抗菌水凝胶;
(4)微凝胶:将去离子水加入到步骤(3)所制得的水凝胶中,去离子水与水凝胶的用量按重量份配比为90:10,再采用高压均质机于30000PSI压力下处理5min,得到控释型纳米纤维素抗菌微凝胶。
通过激光粒度分布仪测得微凝胶的平均粒径范围为2~10μm,放置3天后无明显分层现象。将所得控释型纳米纤维素抗菌微凝胶涂覆在滤纸上干燥后进行抑菌圈测试,以李斯特菌为实验菌种测得第2天抑菌圈为9.1mm,第4天抑菌圈为12.4mm,第6天抑菌圈为13.7mm,表明控释型纳米纤维素抗菌微凝胶能有效控制抗菌剂乳酸链球菌素的释放速度,具有良好的控释抗菌性能。
实施例3
一种控释型纳米纤维素抗菌微凝胶的制备方法,具体操作步骤如下:
(1)制备TEMPO氧化纳米纤维素胶体:采用TEMPO氧化法对漂白阔叶木纤维经机械研磨得到的直径2~20nm、长度0.1~100μm的纳米纤维素进行预处理,得到表面羧基含量为1200mmol/Kg的TEMPO氧化纳米纤维素,然后采用溶剂蒸发法浓缩至质量浓度为3%,真空脱气,得到TEMPO氧化纳米纤维素胶体;
(2)制备乳酸链球菌素溶液:将乳酸链球菌素粉末溶解在浓度为0.01M的盐酸水溶液中,乳酸链球菌素与盐酸水溶液的用量按重量份配比为2:98,溶解后得到质量浓度为2%的乳酸链球菌素溶液;
(3)凝胶化过程:将步骤(1)制得的TEMPO氧化纳米纤维素胶体,置于聚四氟乙烯模具中,再将步骤(2)制得的乳酸链球菌素溶液,用蠕动注射泵以1mL/min的速度沿模具壁滴加到TEMPO氧化纳米纤维素胶体表层,TEMPO氧化纳米纤维素胶体与乳酸链球菌素溶液的用量按重量份配比为80:20,再于4℃下静置24h后形成水凝胶,然后将该水凝胶用等体积的去离子水浸泡冲洗2次,每次1分钟,得到纳米纤维素抗菌水凝胶;
(4)微凝胶:将去离子水加入到步骤(3)所制得的水凝胶中,去离子水与水凝胶的用量按重量份配比为60:40,再采用高速分散机于15000rpm转速下处理5min,得到控释型纳米纤维素抗菌微凝胶。
通过激光粒度分布仪测得微凝胶的平均粒径范围为20~30μm,放置3天后无明显分层现象。将所得控释型纳米纤维素抗菌微凝胶涂覆在滤纸上干燥后进行抑菌圈测试,以李斯特菌为实验菌种测得第2天抑菌圈为10.1mm,第4天抑菌圈为13.3mm,第6天抑菌圈为14.5mm,表明控释型纳米纤维素抗菌微凝胶能有效控制抗菌剂乳酸链球菌素的释放速度,具有良好的控释抗菌性能。

Claims (3)

1.一种控释型纳米纤维素抗菌微凝胶的制备方法,其特征在于,包括如下操作步骤:
(1)制备TEMPO氧化纳米纤维素胶体:采用TEMPO氧化法对纳米纤维素进行预处理,以得到表面羧基含量为600~1200mmol/Kg的TEMPO氧化纳米纤维素,然后采用溶剂蒸发法浓缩至质量浓度为3%,真空脱气,得到TEMPO氧化纳米纤维素胶体;
(2)制备乳酸链球菌素溶液:按重量份配比为2:98的比例将乳酸链球菌素粉末溶解在浓度为0.01M的盐酸水溶液中,得到质量浓度为2%的乳酸链球菌素溶液;
(3)凝胶化过程:将步骤(1)制得的TEMPO氧化纳米纤维素胶体,置于聚四氟乙烯模具中,再将步骤(2)制得的乳酸链球菌素溶液,用蠕动注射泵以1mL/min的速度沿模具壁滴加到TEMPO氧化纳米纤维素胶体表层,TEMPO氧化纳米纤维素胶体与乳酸链球菌素溶液的用量按重量份配比为40~80:20~60,于4℃下静置24~48h后形成水凝胶,然后将该水凝胶用去离子水浸泡冲洗2次,每次1分钟,得到纳米纤维素抗菌水凝胶;
(4)微凝胶:将去离子水加入到步骤(3)制得的水凝胶中,去离子水与水凝胶的用量按重量份配比为60~90:10~40,然后采用机械分散法处理5min,得到控释型纳米纤维素抗菌微凝胶。
2.根据权利要求1所述控释型纳米纤维素抗菌微凝胶的制备方法,其特征在于,步骤(1)中所述的纳米纤维素是指将漂白针叶木纤维、漂白阔叶木纤维或漂白蔗渣纤维采用机械研磨法制备得到直径2~20nm、长度0.1~100μm的纳米纤维素。
3.根据权利要求1或2所述控释型纳米纤维素抗菌微凝胶的制备方法,其特征在于,步骤(4)中所述的机械分散法是指采用超声细胞粉碎机、高速分散机或高压均质机进行处理。
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