CN106317424A - 一种用于高效检测h2o2的磁性复合水凝胶及其制备方法与应用 - Google Patents
一种用于高效检测h2o2的磁性复合水凝胶及其制备方法与应用 Download PDFInfo
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Abstract
本发明公开了一种用于高效检测H2O2的磁性复合水凝胶及其制备方法与应用。该方法步骤为:用丙烯酰氯改性木聚糖,得酰化木聚糖,用热水溶解得酰化木聚糖溶液,加入N‑异丙基丙烯酰胺,丙烯酰胺和交联剂,通氮气,加入光引发剂,混合,置于紫外辐射反应器中,室温下紫外照射,再于室温下封闭放置反应,洗涤润胀;将FeCl3·6H2O与FeSO4·7H2O溶于水中,将酰化木聚糖水凝胶放入此铁盐溶液中,通N2,取出变成黄色的水凝胶浸入氨水中,通N2,得到黑色的水凝胶,洗涤,干燥,得磁性复合水凝胶。本发明制备方法工艺易操作,具有环境友好性;所得水凝胶具有良好的磁敏感性,具有辣根过氧化物酶活性,可用于H2O2的检测。
Description
技术领域
本发明属于天然高分子功能材料制备技术领域,具体涉及一种用于高效检测H2O2的磁性复合水凝胶及其制备方法与应用。
背景技术
近年来,可再生木质纤维素类生物质资源的开发和利用得到了人们的极大重视和关注,被认为是解决石化资源日益枯竭和环境问题的有效途径。半纤维素是农林生物质的主要组分之一,其含量仅次于纤维素,是地球上最丰富、最廉价的可再生资源之一。木聚糖是半纤维素主要的结构形式,主要存在于阔叶木和禾本科植物中。木聚糖本身具有一定的凝胶性,是制备智能水凝胶的良好的原料。
四氧化三铁纳米颗粒(IONPs)具有独特的超顺磁特性,被广泛应用在污水处理、分析检测、生物大分子及细胞分离、药物靶向运输及可控释放、肿瘤磁热治疗、磁共振成像等领域。研究发现IONPs能够模拟辣根过氧化物酶(HRP)活性,催化或氧化氢氧化底物(3,3,5,5-四甲基联苯胺(TMB)、二氨基联苯胺(DAB)、邻苯二胺(OPD))产生颜色变化,其催化活性与HRP类似,依赖于H2O2的浓度、pH和反应温度,催化过程符合米氏动力学和乒乓反应机制,因此能够替代HRP应用在酶联免疫吸附分析(ELISA)。纳米复合水凝胶将纳米材料的优异性质(如,磁性、光热、表面等离子体、催化性等)与智能水凝胶独特的性质(外界刺激如温度、高含水量、pH诱导的相转变)结合起来,可以赋予复合水凝胶优异的性能。磁性复合水凝胶是将磁性材料载入到水凝胶中而得到的。
发明内容
为了克服现有技术的缺点与不足,本发明的首要目的在于提供一种用于高效检测H2O2的磁性复合水凝胶;
本发明的另一目的在于提供上述磁性复合水凝胶的制备方法;
本发明的再一目的在于提供上述磁性复合水凝胶在H2O2检测上的应用。
本发明的目的通过下述技术方案实现。
一种用于高效检测H2O2的磁性复合水凝胶的制备方法,包括如下步骤:
(1)用丙烯酰氯改性木聚糖,得到酰化木聚糖;
(2)将酰化木聚糖用热水溶解,得到酰化木聚糖溶液;
(3)往步骤(2)所得酰化木聚糖溶液中加入N-异丙基丙烯酰胺和丙烯酰胺,再加入交联剂,通氮气鼓泡;加入光引发剂,混合均匀后,冷却至室温,置于紫外辐射反应器中,室温下紫外照射,再于室温下封闭放置反应;反应结束后用去离子水洗涤除去残余反应试剂,得酰化木聚糖基水凝胶;将FeCl3·6H2O与FeSO4·7H2O溶于蒸馏水中,得铁盐溶液;接着将酰化木聚糖基水凝胶胶放入铁盐溶液中浸泡,同时通N2;取出变成黄色的水凝胶,浸入氨水中,同时通N2,得到黑色的水凝胶;均匀切片,干燥,所得固体即为用于高效检测H2O2的磁性复合水凝胶。
优选的,步骤(1)所述酰化木聚糖的具体制备步骤为:将0.6~0.7 g木聚糖溶解在8~12 mL的去离子水中,然后加入15~25 mL二甲基甲酰胺(DMF)搅拌10~15 min,待木聚糖完全溶解后,旋转蒸发除去混合溶液中的水,然后加入0.08~0.12 g LiCl, 0.03~0.04 g 4-二甲氨基吡啶, 10~15 mL 二甲基甲酰胺和2~3 mL丙烯酰氯于65~75℃下反应30~50 min,反应结束后,用75~95wt%的乙醇离心洗涤3~5次,烘干得到酰化木聚糖。
优选的,步骤(1)所述木聚糖为山毛榉木聚糖,分子量为M w =130000 g/mol。
优选的,步骤(2)所述酰化木聚糖溶液的具体制备步骤为:将酰化木聚糖加入温度为70~90 ℃的热水中,搅拌溶解30~60min,得质量浓度为4%~6%的酰化木聚糖溶液,再降温至40~50℃。
优选的,步骤(3)所述交联剂为N,N’-亚甲基双丙烯酰胺;所述光引发剂为安息香二甲醚,所述光引发剂在添加前,先用N-甲基吡咯烷酮(NMP)进行溶解,浓度为2~3wt%。
优选的,步骤(3)所述交联剂的用量为步骤(2)所述酰化木聚糖质量的2.5%~10%;步骤(3)所述丙烯酰胺与步骤(2)所述酰化木聚糖的质量比为(6~12):1;步骤(3)所述N-异丙基丙烯酰胺的用量为步骤(2)所述酰化木聚糖质量的4%~16%;步骤(3)所述光引发剂的用量为步骤(2)所述酰化木聚糖质量的4~6%。
优选的,步骤(3)所述紫外照射的条件:波长为 365 nm、功率为400 W,照射时间为4~8 h;所述放置反应的时间6~10 h;所述通氮气鼓泡的时间为10~30min;所述FeCl3·6H2O与FeSO4·7H2O的摩尔比为(2~4):1;所述酰化木聚糖基水凝胶浸泡于铁盐溶液中的时间为4~8 h;所述氨水的用量为20~30 mL,浓度为25 wt%~28 wt%;所述黄色的水凝胶浸入氨水中通N2的时间为30~60 min;所述干燥的方式为液氮冷冻干燥。
由以上所述的制备方法制得的一种用于高效检测H2O2的磁性复合水凝胶。
以上所述的一种用于高效检测H2O2的磁性复合水凝胶在H2O2检测中的应用。
优选的,所述应用的具体包括如下步骤:将H2O2加入TMB底物显色溶液中,配置一系列的TMB-H2O2显色溶液,将磁性复合水凝胶浸渍于TMB-H2O2显色溶液中,于水浴中反应,溶液变蓝,然后用紫外分光光度计检测TMB-H2O2显色溶液。
优选的,所述TMB-H2O2显色溶液中H2O2的浓度为5uM~200 mM ;所述水浴的温度为37~40℃;所述反应的时间30~60 min;所述检测的紫外检测波长为范围为500~750 nm。
本发明相对于现有技术具有如下的优点及效果:
(1)本发明制备的磁性复合水凝胶,使用的原料为生物质,因木聚糖本身具有可生物降解、生物相容性以及特殊的理化性能,是制备水凝胶理想的原料,本发明能够扩大木聚糖类型半纤维素的工业应用,为半纤维素的高值化利用提供重要的有效途径。
(2)本发明的磁性复合水凝胶具有良好的催化活性,可以用于低浓度H2O2的检测。
(3)本发明的制备方法工艺易操作,容易实现工艺化,具有环境友好性。
附图说明
图1中的a、b、c分别代表本发明交联剂用量为5 wt%、7.5 wt%、10 wt%制得的木聚糖水凝胶的SEM图;
图2中的a、b分别为本发明磁性复合水凝胶的XRD和Raman图;
图3为不同实施例检测相同浓度H2O2的紫外光谱图;
图4为磁性复合水凝胶置于不同H2O2浓度的紫外光谱图。
具体实施方式
下面结合实施例对本发明作进一步详细的描述,但本发明的实施方式不限于此。
实施例1
一种用于高效检测H2O2的磁性复合水凝胶,其制备步骤如下:
将0.6 g山毛榉木聚糖(M w =130000 g/mol)溶解在8 mL的去离子水中,然后加入15 mL二甲基甲酰胺搅拌10 min,待木聚糖完全溶解后,旋转蒸发除去混合溶液中的水,然后加入0.08 g LiCl, 0.03 g 4-二甲氨基吡啶, 10 mL 二甲基甲酰胺和2 mL丙烯酰氯于65 ℃下反应50 min,反应结束后,用75 wt%的乙醇离心洗涤5次,烘干得到酰化木聚糖。
称取2.0 g酰化木聚糖置于三口烧瓶中,加入50 mL去离子水在70℃加热60 min并搅拌使其完全溶解;降温至50℃,然后加入0.08 g的N-异丙基丙烯酰胺(NIPAm)、12 g丙烯酰胺(AM)和0.1 g N,N′-亚甲基双丙烯酰胺(MBA),通氮气以除去溶液中的溶解氧和瓶内空气;然后加入光引发剂安息香二甲醚0.08 g(先将安息香二甲醚0.08 g溶于N-甲基吡咯烷酮中,质量浓度为2%);混合液均匀后,降至室温,倒入石英皿并置于紫外辐射反应器中,设定照射条件为:波长为 365 nm、功率为400 W,室温下照射4 h;达到反应时间后在室温下封闭放置6 h,使其充分聚合和交联;反应完成后,将所得的水凝胶取出,用过量去离子水洗涤以除去水凝胶中残余的反应试剂,然后将水凝胶浸泡在由FeCl3·6H2O与FeSO4·7H2O溶于40 mL蒸馏水中配制所得的盐溶液中4 h,同时通N2 ,水凝胶变黄,然后将变黄的水凝胶放在20 mL氨水溶液(25wt%)中反应30 min,同时通N2 30 min,其中FeCl3·6H2O、FeSO4·7H2O和氨水的摩尔比为2:1:8,反应结束后,将水凝胶浸泡在去离子水中4天,以除去多余的反应物质,然后均匀切块,液氮冷冻干燥即可得到黑色的磁性复合水凝胶。
对所得的水凝胶进行扫描电镜分析,所得的干燥水凝胶的SEM图如图1中的a所示。
对所得的磁性复合水凝胶进行XRD和Raman分析,所得XRD和Raman图谱如图2所示,从图中可以看出水凝胶中生成的磁性颗粒为尖晶石结构的Fe3O4。
将所得的磁性复合水凝胶用于H2O2检测,具体步骤为:将H2O2加入TMB底物显色溶液中,配置一系列的TMB-H2O2显色溶液(5uM~200mM),将本实施例所得磁性复合水凝胶浸渍于TMB-H2O2显色溶液中,于37℃水浴中反应60 min,溶液由无色变蓝色。
将显色反应结束后的蓝色TMB-H2O2显色溶液用紫外分光光度计在波长范围为500~750 nm中检测。对不同交联程度的水凝胶在同一TMB-H2O2显色溶液(50mM)浓度下的催化活性进行分析,如图3所示;对磁性复合水凝胶在不同TMB-H2O2显色溶液(5uM~200mM)中的催化活性进行紫外光谱分析,如图4所示。
经分析,实施例1的磁性复合水凝胶对H2O2有很敏感的检测效果,在5uM~50mM的低浓度范围内,具有较好的显色响应,当H2O2浓度过高,则反应过于剧烈,容易生成白色的沉淀,从而影响紫外吸光度。
实施例2
一种高效检测H2O2的磁性复合水凝胶,其制备步骤如下:
将0.66 g山毛榉木聚糖(M w =130000 g/mol)溶解在10 mL的去离子水中,然后加入20mL二甲基甲酰胺搅拌10 min,待木聚糖完全溶解后,旋转蒸发除去混合溶液中的水,然后加入0.1 g LiCl, 0.035 g 4-二甲氨基吡啶, 12.5 mL 二甲基甲酰胺和2.5 mL丙烯酰氯于70℃下反应40 min,反应结束后,用85 wt%的乙醇离心洗涤4次,烘干得到酰化木聚糖。
称取2.0 g酰化木聚糖置于三口烧瓶中,加入40 mL去离子水在80℃加热45 min并搅拌使其完全溶解;降温至45℃,然后分别加入0. 20g的N-异丙基丙烯酰胺(NIPAm)、18 g丙烯酰胺(AM)和0.15 g N,N′-亚甲基双丙烯酰胺(MBA),通氮气以除去溶液中的溶解氧和瓶内空气;然后加入光引发剂安息香二甲醚0.15 g(先将安息香二甲醚0.15 g溶于N-甲基吡咯烷酮中,质量浓度为2.5%);混合液均匀后,降至室温,倒入石英皿并置于紫外辐射反应器中,设定照射条件为:波长为 365 nm、功率为400 W,室温下照射6 h;达到反应时间后在室温下封闭放置8 h,使其充分聚合和交联;反应完成后,将所得的水凝胶取出用过量去离子水洗涤以除去水凝胶中残余的反应试剂,然后将水凝胶浸泡在由FeCl3·6H2O与FeSO4·7H2O溶于40 mL蒸馏水中配制所得的盐溶液中6 h,同时通N2,水凝胶变黄,然后将变黄的水凝胶放在25 mL氨水溶液(26.5 wt%)中反应30 min,同时通N2 45 min,其中FeCl3·6H2O、FeSO4·7H2O和氨水的摩尔比为3:1:8,反应结束后,将水凝胶浸泡在去离子水中4天,以除去多余的反应物质,然后均匀切块,液氮冷冻干燥即可得到黑色的磁性复合水凝胶。
将所得的磁性复合水凝胶用于H2O2检测,具体步骤为:将H2O2加入TMB底物显色溶液中,配置一系列的TMB-H2O2显色溶液(5 uM~200 mM),将所得磁性复合水凝胶浸渍于TMB-H2O2显色溶液中,于38.5℃水浴中反应45 min,溶液由无色变为蓝色。
对所得的水凝胶进行扫描电镜分析,所得的干燥水凝胶的SEM图如图1中的b所示。
将显色反应结束后的蓝色TMB-H2O2显色溶液用紫外分光光度计在波长为500~750nm中检测。对不同交联程度的水凝胶在同一TMB-H2O2显色溶液(50mM)浓度下的催化活性进行分析,如图3所示。
研究表明,本实施例所制备的磁性复合水凝胶交联度更大,结构更为紧密,对H2O2的催化活性最高。
实施例3
一种高效检测H2O2的磁性复合水凝胶,其制备步骤如下:
将0.72 g山毛榉木聚糖(M w =130000 g/mol)溶解在12 mL的去离子水中,然后加入25mL二甲基甲酰胺搅拌15 min,待木聚糖完全溶解后,旋转蒸发除去混合溶液中的水,然后加入0.12 g LiCl, 0.04 g 4-二甲氨基吡啶, 25 mL 二甲基甲酰胺和3 mL丙烯酰氯于75℃下反应30 min,反应结束后,用95 wt%的乙醇离心洗涤3次,烘干得到酰化木聚糖。
称取2.0 g酰化木聚糖置于三口烧瓶中,加入30 mL去离子水在90℃加热30 min并搅拌使其完全溶解;降温至40℃,然后加入0.32 g的N-异丙基丙烯酰胺(NIPAm)、24 g丙烯酰胺(AM)和0.2 g N,N′-亚甲基双丙烯酰胺(MBA),通氮气鼓泡以除去溶液中的溶解氧和瓶内空气;然后加入光引发剂安息香二甲醚0.12 g(先将安息香二甲醚0.12 g溶于N-甲基吡咯烷酮中,质量浓度为3%);混合液均匀后,降至室温,倒入石英皿并置于紫外辐射反应器中,设定照射条件为:波长为 365 nm、功率为400 W,室温下照射8 h;达到反应时间后在室温下封闭放置10 h,使其充分聚合和交联;反应完成后,将所得的水凝胶取出用过量去离子水洗涤以除去水凝胶中残余的反应试剂,然后将水凝胶浸泡在由FeCl3·6H2O与的FeSO4·7H2O溶于40 mL蒸馏水中配制所得的盐溶液中8 h,同时通N2 ,水凝胶变黄,然后将变黄的水凝胶放在30 mL氨水溶液(28 wt%)中反应30 min,同时通N2 60 min,其中FeCl3·6H2O、FeSO4·7H2O和氨水的摩尔比为4:1:8,反应结束后,将水凝胶浸泡在去离子水中4天,以除去多余的反应物质,然后均匀切块,液氮冷冻干燥即可得到黑色的磁性复合水凝胶。
将所得的磁性复合水凝胶用于H2O2检测,具体步骤为:将H2O2加入TMB底物显色溶液中,配置一系列的TMB-H2O2显色溶液(5 uM~200 mM),将所得磁性复合水凝胶浸渍于TMB-H2O2显色溶液中,于40℃水浴中反应60 min,溶液由无色变为蓝色。
对所得的水凝胶进行扫描电镜分析,所得的干燥水凝胶的SEM图如图1中的c所示。
将显色反应结束后的蓝色TMB-H2O2显色溶液用紫外分光光度计在波长为500~750nm中检测。对不同交联程度的水凝胶在相同TMB-H2O2显色溶液(50mM)浓度下的催化活性进行分析,如图3所示。
研究表明,本实施例所制备的磁性复合水凝胶交联度更大,结构更为紧密,对H2O2的催化活性较实施例2和实施例1的磁性复合水凝胶催化效率低。这说明磁性复合水凝胶的催化效率受水凝胶的交联度影响。
上述实施例2和实施例3的图2和图4和实施例1基本相同,不一一说明。
上述实施例为本发明较佳的实施方式,但本发明的实施方式并不受上述实施例的限制,其他的任何未背离本发明的精神实质与原理下所作的改变、修饰、替代、组合、简化,均应为等效的置换方式,都包含在本发明的保护范围之内。
Claims (10)
1.一种用于高效检测H2O2的磁性复合水凝胶的制备方法,其特征在于,包括如下步骤:
(1)用丙烯酰氯改性木聚糖,得到酰化木聚糖;
(2)将酰化木聚糖用热水溶解,得到酰化木聚糖溶液;
(3)往步骤(2)所得酰化木聚糖溶液中加入N-异丙基丙烯酰胺和丙烯酰胺,再加入交联剂,通氮气鼓泡;加入光引发剂,混合均匀后,冷却至室温,置于紫外辐射反应器中,室温下紫外照射,再于室温下封闭放置反应;反应结束后用去离子水洗涤除去残余反应试剂,得酰化木聚糖基水凝胶;将FeCl3·6H2O与FeSO4·7H2O溶于蒸馏水中,得铁盐溶液;接着将酰化木聚糖基水凝胶放入铁盐溶液中浸泡,同时通N2;取出变成黄色的水凝胶,浸入氨水中,同时通N2,得到黑色的水凝胶;均匀切片,干燥,所得固体即为用于高效检测H2O2的磁性复合水凝胶。
2.根据权利要求1所述的一种用于高效检测H2O2的磁性复合水凝胶的制备方法,其特征在于,步骤(1)所述酰化木聚糖的具体制备步骤为:将0.6~0.72 g木聚糖溶解在8~12 mL的去离子水中,然后加入15~25 mL二甲基甲酰胺搅拌10~15 min,待木聚糖完全溶解后,旋转蒸发除去混合溶液中的水,然后加入0.08~0.12 g LiCl, 0.03~0.04 g 4-二甲氨基吡啶,10~15 mL 二甲基甲酰胺和2~3 mL丙烯酰氯于65~75℃下反应30~50 min,反应结束后,用75~95wt%的乙醇离心洗涤3~5次,烘干得到酰化木聚糖。
3.根据权利要求1所述的一种用于高效检测H2O2的磁性复合水凝胶的制备方法,其特征在于,步骤(2)所述酰化木聚糖溶液的具体制备步骤为:将酰化木聚糖加入温度为70~90℃的热水中,搅拌溶解30~60min,得质量浓度为4%~6%的酰化木聚糖溶液,再降温至40~50℃。
4.根据权利要求1所述的一种用于高效检测H2O2的磁性复合水凝胶的制备方法,其特征在于,步骤(3)所述交联剂为N,N’-亚甲基双丙烯酰胺;所述光引发剂为安息香二甲醚,所述光引发剂在添加前,先用N-甲基吡咯烷酮进行溶解,所得溶液浓度为2~3wt%。
5.根据权利要求1所述的一种用于高效检测H2O2的磁性复合水凝胶的制备方法,其特征在于,步骤(3)所述交联剂的用量为步骤(2)所述酰化木聚糖质量的5%~10%;步骤(3)所述丙烯酰胺与步骤(2)所述酰化木聚糖的质量比为(6~12):1;步骤(3)所述N-异丙基丙烯酰胺的用量为步骤(2)所述酰化木聚糖质量的4%~16%;步骤(3)所述光引发剂的用量为步骤(2)所述酰化木聚糖质量的4~6%。
6.根据权利要求1所述的一种用于高效检测H2O2的磁性复合水凝胶的制备方法,其特征在于,步骤(3)所述通氮气鼓泡的时间为10~30min;所述紫外照射的条件:波长为 365 nm、功率为400 W,照射时间为4-8 h;所述放置反应的时间为6~10 h;所述FeCl3·6H2O与FeSO4·7H2O的摩尔比为(2~4):1;所述木聚糖水凝胶浸泡于铁盐溶液中的时间为4~8 h;所述氨水的用量为20~30 mL,浓度为25~28wt%;所述黄色的水凝胶浸入氨水中通N2的时间为30~60 min;所述干燥的方式为液氮冷冻干燥。
7.由权利要求1~6任一项所述的制备方法制得的一种用于高效检测H2O2的磁性复合水凝胶。
8.权利要求7所述的一种用于高效检测H2O2的磁性复合水凝胶在H2O2检测中的应用。
9.根据权利要求8所述的应用,其特征在于,具体包括如下步骤:将H2O2加入TMB底物显色溶液中,配置TMB-H2O2显色溶液,将磁性复合水凝胶浸渍于TMB-H2O2显色溶液中,于水浴中反应,溶液变蓝,然后用紫外分光光度计检测TMB-H2O2显色溶液。
10.根据权利要求9所述的应用,其特征在于,所述TMB-H2O2显色溶液中H2O2的浓度为5uM~200mM;所述水浴的温度为37~40℃;所述反应的时间为30~60 min;所述检测的紫外检测波长为500~750 nm。
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CN111498875A (zh) * | 2020-04-17 | 2020-08-07 | 中南大学 | 一种利用盐湖镁资源制备氢氧化镁或轻质氧化镁的方法 |
CN111498875B (zh) * | 2020-04-17 | 2021-04-13 | 中南大学 | 一种利用盐湖镁资源制备氢氧化镁或轻质氧化镁的方法 |
CN112362653A (zh) * | 2020-10-29 | 2021-02-12 | 湖南久日新材料有限公司 | 一种光引发剂低氯含量的检测方法 |
CN112362653B (zh) * | 2020-10-29 | 2024-02-27 | 湖南久日新材料有限公司 | 一种光引发剂低氯含量的检测方法 |
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