CN115505386A - 基于bpei和sds的四环素检测材料及其制备方法和应用 - Google Patents
基于bpei和sds的四环素检测材料及其制备方法和应用 Download PDFInfo
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Abstract
本发明公开了基于BPEI和SDS的四环素检测材料及其制备方法和应用,解决了现有技术中铜纳米团簇的荧光变化程度低、检出限高以及特异性和抗干扰性能较差的技术问题。基于BPEI和SDS的四环素检测材料包括铜纳米团簇和附着于铜纳米团簇上的十二烷基硫酸钠;所述铜纳米团簇以聚乙烯亚胺为模板制备得到。制备方法包括以下步骤:(1)获取包括Cu2+、聚乙烯亚胺和抗坏血酸的第一混合液;(2)第一混合液反应一段时间即得到铜纳米团簇分散液;(4)将铜纳米团簇分散液和十二烷基硫酸钠溶液混合,即得到检测材料。经验证,本发明的基于BPEI和SDS的四环素检测材料及其制备方法和应用的实用性强,非常适合于在实际样品的四环素检测中推广使用。
Description
技术领域
本发明涉及四环素检测的技术领域,具体而言,涉及基于BPEI和SDS的四环素检测材料及其制备方法和应用。
背景技术
四环素(TC)长期以来一直用于治疗细菌感染。但是,四环素的过度使用会对环境中的植物以及微生物的成长造成威胁。因此,开发一种高灵敏度、高选择性的检测方法显得尤为重要。之前已经有报道过的多种方法,包括高效液相法、免疫测定、化学发光、液相色谱质谱和荧光法。
荧光法因其低成本、高灵敏度、高选择性等优点已广泛应用于实际样品中物质的检测。近些年关于各种纳米材料的荧光探针,包括量子点、碳点和纳米团簇已被广泛研究,其中,纳米团簇包括银纳米团簇、金纳米团簇以及铜纳米团簇(CuNCs)。目前,金纳米团簇和银纳米团簇研究较早且较为广泛。然而,与贵金属金和银相比,金属铜价格相对便宜且用于合成CuNCs的前体相对丰富、易于获得,因而使得CuNCs在四环素荧光探针的研究中有很大的发展空间。
但是,CuNCs相对于其他的金属纳米团簇,不仅其本身的荧光强度相对过低,并且在实践中发现,CuNCs在对四环素进行检测时的荧光变化程度、检出限高、特异性和抗干扰性能难以满足实际样品中对四环素的检测要求。
发明内容
本发明的主要目的在于提供基于BPEI和SDS的四环素检测材料及其制备方法和应用,以解决现有技术中铜纳米团簇的荧光变化程度低、检出限高以及特异性和抗干扰性能较差的技术问题。
为了实现上述目的,根据本发明的第一个方面,提供了基于BPEI和SDS的四环素检测材料,技术方案如下:
基于BPEI和SDS的四环素检测材料,包括铜纳米团簇和附着于铜纳米团簇上的十二烷基硫酸钠;所述铜纳米团簇以聚乙烯亚胺为模板制备得到。
作为本发明第一方面的进一步改进:所述检测材料的粒度为1.8~3.4nm;所述铜纳米团簇的粒度为0.6~2.4nm。
作为本发明第一方面的进一步改进:所述检测材料的FT-IR光谱在1575cm-1、1472cm-1、2918cm-1和2848cm-1处处具有特征峰,在3100~3600cm-1处具有宽频带的伸缩振动。
作为本发明第一方面的进一步改进:所述检测材料的XPS宽谱图在284.94eV、932.27eV、399.79eV和531.65eV处具有特征峰。
作为本发明第一方面的进一步改进:所述检测材料的激发波长为360nm,发射波长为430nm。
为了实现上述目的,根据本发明的第二个方面,提供了基于BPEI和SDS的四环素检测材料的制备方法,技术方案如下:
上述第一方面所述基于BPEI和SDS的四环素检测材料的制备方法,包括以下步骤:
(1)获取包括Cu2+、聚乙烯亚胺和抗坏血酸的第一混合液;
(2)第一混合液反应一段时间即得到铜纳米团簇分散液;
(4)将铜纳米团簇分散液和十二烷基硫酸钠溶液混合,即得到检测材料。
作为本发明第二方面的进一步改进:所述第一混合液中Cu2+、聚乙烯亚胺和抗坏血酸的摩尔比为1:(8~12):(43~52)。
作为本发明第二方面的进一步改进:第一混合液在18~35℃下震荡反应1-3天即得到铜纳米团簇分散液。
作为本发明第二方面的进一步改进:第一混合液的pH为8~12。
为了实现上述目的,根据本发明的第三个方面,提供了四环素检测方法,技术方案如下:
四环素的检测方法,采用上述第一方面所述检测材料,或采用上述第二方面所述的制备方法制备得到的检测材料。
经验证,本发明的四环素的检测材料和检测方法利用由新型的聚乙烯亚胺(BPEI)和十二烷基硫酸钠(SDS)进行改性的铜纳米团簇对四环素进行检测,成功使得荧光变化程度更明显,并且能够在众多抗生素中特异性识别四环素,并且抗干扰性能优异,线性范围宽,检出限低,非常适合于对四环素进行检测。同时,上述检测材料的制备工艺简单可控,生产效率高。由此可见,本发明的基于BPEI和SDS的四环素检测材料及其制备方法和应用的实用性强,非常适合于在实际样品的四环素检测中推广使用。
下面结合附图和具体实施方式对本发明做进一步的说明。本发明附加的方面和优点将在下面的描述中部分给出,部分将从下面的描述中变得明显,或通过本发明的实践了解到。
附图说明
构成本发明的一部分的附图用来辅助对本发明的理解,附图中所提供的内容及其在本发明中有关的说明可用于解释本发明,但不构成对本发明的不当限定。在附图中:
图1为BPEI-CuNCs-SDS分散液的F0-F随SDS浓度的变化曲线。
图2为检测材料的荧光强度随表面活性剂种类的变化曲线。
图3为四环素溶液的紫外-可见吸收光谱以及BPEI-CuNCs-SDS分散液的荧光激发光谱和发射光谱。
图4为BPEI-CuNCs-SDS分散液和BPEI-CuNCs-SDS+TC的荧光衰减光谱。
图5为BPEI-CuNCs-SDS分散液的F0-F随四环素溶液的pH的变化曲线。
图6为BPEI-CuNCs-SDS分散液的F0-F随反应时间的变化曲线。
图7为BPEI-CuNCs-SDS分散液的F0/F随抗生素种类的变化曲线。
图8为BPEI-CuNCs-SDS分散液的抗干扰性能测试结果。
图9为BPEI-CuNCs-SDS分散液测试不同浓度的四环素溶液的连续荧光光谱图。
图10为BPEI-CuNCs-SDS分散液的F0/F随四环素浓度的变化曲线。
图11为BPEI-CuNCs-SDS分散液的F0/F与四环素浓度(0.5~60μmol/L)的线性校准图。
图12为BPEI-CuNCs-SDS分散液的F0/F与四环素浓度(60~140μmol/L)的线性校准图。
图13为BPEI-CuNCs粉末的TEM照片。
图14为BPEI-CuNCs粉末的粒径分布直方图。
图15为BPEI-CuNCs-SDS粉末的TEM照片。
图16为BPEI-CuNCs-SDS粉末的粒径分布直方图。
图17为SDS、BPEI-CuNCs粉末和BPEI-CuNCs-SDS粉末的红外光谱(FT-IR)。
图18为BPEI-CuNCs-SDS粉末的XPS全谱图。
图19为BPEI-CuNCs-SDS粉末在Cu2p的XPS宽谱图。
具体实施方式
下面结合附图对本发明进行清楚、完整的说明。本领域普通技术人员在基于这些说明的情况下将能够实现本发明。在结合附图对本发明进行说明前,需要特别指出的是:
本发明中在包括下述说明在内的各部分中所提供的技术方案和技术特征,在不冲突的情况下,这些技术方案和技术特征可以相互组合。
此外,下述说明中涉及到的本发明的实施例通常仅是本发明一部分的实施例,而不是全部的实施例。因此,基于本发明中的实施例,本领域普通技术人员在没有做出创造性劳动的前提下所获得的所有其他实施例,都应当属于本发明保护的范围。
关于本发明中术语和单位。本发明的说明书和权利要求书及有关的部分中的术语“包括”、“具有”以及它们的任何变形,意图在于覆盖不排他的包含。
本发明的基于BPEI和SDS的四环素检测材料的制备方法的具体实施方式为包括以下步骤:
(1)获取包括Cu2+、BPEI和抗坏血酸(AA)的第一混合液;
所述第一混合液中Cu2+、AA和BPEI的摩尔比为1:(8~12):(43~52),可以但是不限于取值为1:8:43、1:9:45、1:10:48、1:11:50、1:12:52中的任意一个。
Cu2+由硝酸铜、氯化铜或硫酸铜提供。
第一混合液的pH为8~12,可以但是不限于取值为8、9、10、11、12中的任意一个。
(2)第一混合液在18~35℃下震荡反应1-3天即得到铜纳米团簇分散液(即BPEI-CuNCs分散液);
对BPEI-CuNCs分散液进行冷冻干燥,即可得到BPEI-CuNCs粉末。
(4)在将BPEI-CuNCs分散液和SDS溶液混合后进行超声处理以使溶液均质化,形成BPEI-CuNCs-SDS的分散液,即得到检测材料。
本发明的基于BPEI和SDS的四环素检测材料的具体实施方式为由上述的制备方法制备得到,具体包括BPEI-CuNCs-SDS粉末或BPEI-CuNCs-SDS的分散液,BPEI-CuNCs-SDS粉末由BPEI-CuNCs-SDS的分散液经冷冻干燥得到。所述BPEI-CuNCs-SDS包括铜纳米团簇和附着于铜纳米团簇上的SDS,并且所述铜纳米团簇以BPEI为模板制备得到。
对BPEI-CuNCs-SDS粉末和BPEI-CuNCs粉末进行表征得到:BPEI-CuNCs-SDS的粒度为1.8-3.4nm;BPEI-CuNCs的粒度为0.6~2.4nm;BPEI-CuNCs-SDS的FT-IR光谱在1575cm-1、1472cm-1、2918cm-1和2848cm-1处处具有特征峰,在3100~3600cm-1处具有宽频带的伸缩振动;BPEI-CuNCs-SDS的XPS宽谱图在284.94eV、932.27eV、399.79eV和531.65eV处具有特征峰;BPEI-CuNCs-SDS的激发波长为360nm,发射波长为430nm。
以下通过具体的表征和试验来说明本发明的有益效果。
图1为BPEI-CuNCs-SDS分散液的F0-F随SDS浓度的变化曲线。其中,F0为BPEI-CuNCs-SDS分散液的荧光强度,F为BPEI-CuNCs-SDS+TC(即BPEI-CuNCs-SDS分散液和四环素溶液的混合物)的荧光强度。其它的制备工艺参数为:第一混合液中Cu2+、AA和BPEI的摩尔比为1:10:48.3,将1.73mL硫酸铜溶液(浓度为10mmol/L)、1.73mLAA溶液(浓度为100mmol/L)、36mgBPEI和100mL水混合,然后采用1mol/L的HCl调节pH为10,即得到第一混合液;震荡时间为2天;BPEI-CuNCs-SDS分散液由BPEI-CuNCs分散液和SDS溶液按照体积比为1:1混合而成,BPEI-CuNCs-SDS分散液的体积为200μL。
如图1所示,SDS溶液的最佳浓度为0.6mg/mL。
图2为检测材料的荧光强度随表面活性剂种类的变化曲线。
从图2可以看出,将不同类型的表面活性剂如阳离子表面活性剂(CTAB和CTAC)、阴离子表面活性剂(SDS和SDBS)和非离子表面活性剂(Tween-20和Tween-80)加入到BPEI-CuNCs分散液中,荧光强度相比于BPEI-CuNCs分散液分别增强了1.02、1.67、1.98、1.83、1.49和1.38倍,可见,虽然这些表面活性剂对BPEI-CuNCs都有明显的增强荧光效果,但是SDS的荧光增强效果最佳。其中,每种表面活性剂的浓度相同,CTAB为溴代十六烷基三甲胺,CTAC为十六烷基三甲基氯化铵,SDBS为十二烷基苯磺酸钠,Tween-20为吐温-20,Tween-80为吐温-80。
图3为四环素溶液的紫外-可见吸收光谱以及BPEI-CuNCs-SDS分散液的荧光激发光谱和发射光谱。
导致荧光猝灭的机制主要有光致电子转移(PET)、荧光共振能量转移(FRET)和内滤效应(IFE)。如图3所示,四环素溶液的紫外-可见吸收光谱在300~400nm范围内有吸收峰,与BPEI-CuNCs-SDS分散液的激发光谱重叠,说明四环素诱导的荧光猝灭机制可能是荧光共振能量转移(FRET)或内滤效应(IFE)。由图3可以得到BPEI-CuNCs-SDS分散液的荧光激发光谱的波峰在360nm处,发射光谱的波峰在430nm处。
图4为BPEI-CuNCs-SDS分散液和BPEI-CuNCs-SDS+TC的荧光衰减光谱。
对图4用双指数函数拟合荧光衰减得到BPEI-CuNCs-SDS分散液的平均荧光寿命为3.55ns,BPEI-CuNCs-SDS+TC的平均荧光寿命为3.12ns,因此,BPEI-CuNCs-SDS的荧光被四环素通过FRET机制有效地猝灭。另外,BPEI-CuNCs-SDS的能量会转移到BPEI-CuNCs-SDS和四环素的复合物中,从而使BPEI-CuNCs-SDS的荧光猝灭。
图5为BPEI-CuNCs-SDS分散液的F0-F随四环素溶液的pH的变化曲线。图6为BPEI-CuNCs-SDS分散液的F0-F随反应时间的变化曲线。其中,四环素溶液的pH采用pH为2~7的PBS缓冲液来调节。F0为BPEI-CuNCs-SDS分散液的荧光强度,F为BPEI-CuNCs-SDS+TC的荧光强度。
如图5所示,当四环素溶液的pH为3时,F0-F具有最大值。如图6所示,当BPEI-CuNCs-SDS分散液与四环素溶液的反应时间在5~60min内时,F0-F几乎无变化,为了节约时间,优选反应时间为5min。
图7为BPEI-CuNCs-SDS分散液的F0/F随抗生素种类的变化曲线。F0为BPEI-CuNCs-SDS分散液的荧光强度,F为BPEI-CuNCs-SDS+抗生素的荧光强度。
如图7所示,在四环素、土霉素、强力霉素、金霉素、阿莫西林、诺氟沙星、氧氟沙星、氯苄西林、阿奇霉素、罗红霉素、头孢氨苄中,BPEI-CuNCs-SDS分散液测试四环素的F0/F最大,说明BPEI-CuNCs-SDS能够特异性识别四环素。
图8为BPEI-CuNCs-SDS分散液的抗干扰性能测试结果。F0为BPEI-CuNCs-SDS分散液的荧光强度,F为BPEI-CuNCs-SDS+TC+干扰离子的荧光强度。
如图8所示,在金属离子和阴离子共存的情况下,BPEI-CuNCs-SDS分散液的测试四环素溶液的F0/F几乎没有变化,表明BPEI-CuNCs-SDS对四环素具有良好的抗干扰能力,可以满足实际应用中的要求。
图9为BPEI-CuNCs-SDS分散液测试不同浓度的四环素溶液的连续荧光光谱图。图10为BPEI-CuNCs-SDS分散液的F0/F随四环素浓度的变化曲线。图11为BPEI-CuNCs-SDS分散液的F0/F与四环素浓度(0.5~60μmol/L)的线性校准图。图12为BPEI-CuNCs-SDS分散液的F0/F与四环素浓度(60~140μmol/L)的线性校准图。F0为BPEI-CuNCs-SDS分散液的荧光强度,F为BPEI-CuNCs-SDS+TC的荧光强度。
如图9所示,随着四环素浓度的提升,BPEI-CuNCs-SDS+TC的荧光强度逐渐减小。如图10所示,F0/F与四环素的浓度呈现良好的线性关系,且存在两段线性关系。如图11所示,当四环素浓度为0.5~60μmol/L时,回归方程为:y=0.0352x+0.9328,R2=0.9974;如图12所示,当四环素浓度为60~140μmol/L时,回归方程为:y=0.1279x-4.7842,R2=0.9925。BPEI-CuNCs-SDS分散液检测四环素浓度的检出限为105.3nmol/L,远小于现有技术中的检测限,具有突出优势。
选取西南交通大学校园湖水以及实验室自来水作为实际样品,离心10min后用0.45μm的滤膜过滤,然后向水样中加入四环素形成不同浓度的四环素溶液,采用BPEI-CuNCs-SDS分散液和上述的回归方程测试水样中四环素的浓度。表1显示了BPEI-CuNCs-SDS分散液对实际水样中四环素的检测结果。
表1
从表1可以看出,四环素的回收率在90.1~109.5%范围内,且相对标准偏差(RSD)均小于8.9%,说明BPEI-CuNCs-SDS在实际水样中检测四环素的准确性较高,可以构建新型荧光探针用于快速检测真实样品中的四环素,具有极大的实际应用价值。
在上述的荧光测试中,四环素溶液和抗生素溶液的体积为60μL;四环素溶液浓度作为定量时的浓度为2mmol/L;干扰离子的体积为60μL,浓度为500μmol/L;采用PBS缓冲液将待进行荧光检测的液体体积定容至1.5mL。
图13为BPEI-CuNCs粉末的TEM照片。图14为BPEI-CuNCs粉末的粒径分布直方图。图15为BPEI-CuNCs-SDS粉末的TEM照片。图16为BPEI-CuNCs-SDS粉末的粒径分布直方图。
如图13-16所示,BPEI-CuNCs粉末和BPEI-CuNCs-SDS粉末均由均匀的球形纳米颗粒构成,其中,BPEI-CuNCs粉末由粒度为0.6~2.4nm的纳米颗粒构成,平均粒度为1.54nm,BPEI-CuNCs-SDS粉末由粒度为1.8~3.4nm的纳米颗粒构成,平均粒度为2.62nm。
图17为SDS、BPEI-CuNCs粉末和BPEI-CuNCs-SDS粉末的红外光谱(FT-IR)。
如图17所示,BPEI-CuNCs-SDS粉末的FT-IR光谱显示出SDS的特征峰,说明SDS成功修饰在BPEI-CuNCs表面。在BPEI-CuNCs-SDS粉末的FT-IR光谱中,在1575cm-1处得到的特征峰对应于-NH2和-NH-的弯曲振动,在1472cm-1处的特征峰对应于C-H的收缩振动,在3100~3600cm-1处的宽频带对应于O-H和N-H基团的伸缩振动,在2918cm-1和2848cm-1处的较强特征峰对应于-CH2基团的对称和反对称振动。
图18为BPEI-CuNCs-SDS粉末的XPS全谱图。
如图18所示,在284.94、932.27、399.79和531.65eV处的特征峰分别归因于C1s、Cu2p、N1s和O1s,计算得到BPEI-CuNCs-SDS粉末中C、Cu、N、O的质量分数分别为53.05、0.21、16.80和29.94wt%。
图19为BPEI-CuNCs-SDS粉末在Cu2p的XPS宽谱图。
在图19中,在结合能位于932.3eV和952.1eV处的两个特征峰分别对应于Cu2p3/2和Cu2p1/2,表明BPEI-CuNCs-SDS中铜的价态为零价。
上述表征和性能测试中,荧光光谱在FLS1000稳态/瞬态荧光光谱仪(英国爱丁堡公司)上进行,激发和发射光谱的狭缝宽度为1.5nm。使用PHS-3W pH计(中国上海云赛公司)测量溶液的pH值。K-Alpha+X射线光电子能谱(美国赛默飞公司)用于测试材料的元素组成及价态。USB-2000UV-vis光谱仪(美国海洋光学公司)用于测量材料的吸收特性。使用FEITecnai G2 F20透射电子显微镜(美国)测试所制备的材料的形貌特征。FT-IR采用美国珀金莱默傅里叶变换红外光谱。
以上对本发明的有关内容进行了说明。本领域普通技术人员在基于这些说明的情况下将能够实现本发明。基于本发明的上述内容,本领域普通技术人员在没有做出创造性劳动的前提下所获得的所有其他实施例,都应当属于本发明保护的范围。
Claims (10)
1.基于BPEI和SDS的四环素检测材料,其特征在于:包括铜纳米团簇和附着于铜纳米团簇上的十二烷基硫酸钠;所述铜纳米团簇以聚乙烯亚胺为模板制备得到。
2.如权利要求1所述的基于BPEI和SDS的四环素检测材料,其特征在于:所述检测材料的粒度为1.8~3.4nm;所述铜纳米团簇的粒度为0.6~2.4nm。
3.如权利要求1所述的基于BPEI和SDS的四环素检测材料,其特征在于:所述检测材料的FT-IR光谱在1575cm-1、1472cm-1、2918cm-1和2848cm-1处处具有特征峰,在3100~3600cm-1处具有宽频带的伸缩振动。
4.如权利要求1所述的基于BPEI和SDS的四环素检测材料,其特征在于:所述检测材料的XPS宽谱图在284.94eV、932.27eV、399.79eV和531.65eV处具有特征峰。
5.如权利要求1所述的基于BPEI和SDS的四环素检测材料,其特征在于:所述检测材料的激发波长为360nm,发射波长为430nm。
6.权利要求1-5所述基于BPEI和SDS的四环素检测材料的制备方法,包括以下步骤:
(1)获取包括Cu2+、聚乙烯亚胺和抗坏血酸的第一混合液;
(2)第一混合液反应一段时间即得到铜纳米团簇分散液;
(4)将铜纳米团簇分散液和十二烷基硫酸钠溶液混合,即得到检测材料。
7.如权利要求6所述的制备方法,其特征在于:所述第一混合液中Cu2+、聚乙烯亚胺和抗坏血酸的摩尔比为1:(8~12):(43~52)。
8.如权利要求6所述的制备方法,其特征在于:第一混合液在18~35℃下震荡反应1-3天即得到铜纳米团簇分散液。
9.如权利要求6所述的制备方法,其特征在于:第一混合液的pH为8~12。
10.四环素的检测方法,采用权利要求1-5之一所述检测材料,或采用权利要求6-9之一所述的制备方法制备得到的检测材料。
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