CN108659833A - 一种黄色荧光碳点及其制备方法和应用 - Google Patents
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- LISFMEBWQUVKPJ-UHFFFAOYSA-N quinolin-2-ol Chemical compound C1=CC=C2NC(=O)C=CC2=C1 LISFMEBWQUVKPJ-UHFFFAOYSA-N 0.000 claims abstract description 33
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Abstract
本发明公开了一种发射黄色荧光的碳点及其制备方法,以及该碳点用于免标记检测喹诺酮类抗生素。所述碳点为一种以邻苯二胺和氨基丁酸为原料,通过一步水热合成法制备的黄色荧光碳点。碳点的制备方法简便,制得的碳点光学性质稳定,水溶性好,分散性好。由于碳点独特的表面结构,以其作为荧光探针能够免标记地、快速灵敏地检测喹诺酮类抗生素。当喹诺酮类抗生素存在时,碳点的荧光被喹诺酮类抗生素有效淬灭,根据荧光的变化程度可以检测出样品中喹诺酮类抗生素的含量。当本发明提供的检测喹诺酮类抗生素的方法与传统方法相比,简便快速,无需额外修饰标记物,可直接检测样品,在实际应用中具有更多的优势。
Description
技术领域
本发明涉及荧光碳点,具体涉及一种黄色荧光碳点及其制备方法,以及该碳点用于喹诺酮类抗生素的免标记检测。
背景技术
喹诺酮类抗生素是人工合成的第三代含4-喹诺酮基本结构的用于预防和治疗致病细菌感染抗生素药物。喹诺酮类以细菌的脱氧核糖核酸(DNA)为靶,妨碍DNA回旋酶,进一步造成细菌DNA的不可逆损害,破坏细菌的正常代谢、繁殖,从而达到抗菌效果。由于其渗透力强、半衰期较长、对环境和食品中的革兰氏阳性菌和阴性菌均表现出广泛的抗菌活性,因此,在人类和动物中被广泛用于治疗呼吸道及皮肤感染性疾病等。但因其日益增多的用量,现已成为水体等环境的主要污染物,并且会残留于食物中。这会直接影响人类健康、诱导产生耐药菌、影响生态系统。因此,开发一种有效,简单,低成本的监测工具来直接检测这类抗生素的残留量具有重要意义。
据目前的研究报道,对喹诺酮类抗生素的检测主要集中于两类方法。一类是仪器分析,例如使用HPLC-MS、GC-MS等分析仪器;或者利用基于抗原与抗体相互作用的免疫方法例如ELISAs。虽然这些方法有许多优点,例如检测样品高通量、选择性好、灵敏度高,但是仍然遭受着样品前处理步骤复杂,需要标记特定的靶向物质,需要昂贵的仪器,检测进程耗时长等问题。因此,开发一种简便快速,选择性好,灵敏度高的分析技术检测喹诺酮类抗生素具有重要意义。
荧光分析法由于灵敏度高、选择性好、操作简单快捷等检测优势得到广泛应用。目前,利用化学性质优异、毒性低、生物相容性好的碳点作为荧光探针已经在生化传感、生物成像、环境分析等领域得到了良好的应用。但是,利用简便的制备方法,可控制备具有特定结构的荧光碳点,并将其应用于靶向分子的检测仍然具有挑战性。
发明内容
为解决上述技术问题,本发明的目的在于提供一种结构性质优异的荧光碳点及其制备方法,碳点的制备方法简便、原料廉价易得;所制备的碳点表面化学结构优异,可应用于免标记检测喹诺酮类抗生素,并展现出出良好的选择性和灵敏度。
本发明提供的一种黄色荧光碳点的制备方法,包括以下步骤:
1)、将邻苯二胺和氨基丁酸置于玻璃烧杯中,加入去离子水中,充分搅拌后超声溶解15-20分钟,邻苯二胺和氨基丁酸的质量比为:0.15-0.62∶0.162-0.648;
2)、将反应混合物转移至水热反应釜中,并置于烘箱内,160℃反应8-10h,得到棕色溶液;
3)、取出水热反应釜,自然冷却,将反应产物加入去离子水稀释10-20倍,在13000r/m转速下离心15min,去除不溶物得到澄清棕黄色的溶液,用0.22μm滤膜过滤后得到纯净的碳点的水溶液;
4)、将上述碳点水溶液冷冻干燥后得到碳点固体。
上述方法制备的碳点荧光性质稳定,在紫外灯照射下发出明亮的黄色荧光;具有良好的水溶性和分散性,通过AFM表征可看出其形貌为单分散无定型颗粒。所述碳点制备方法简便,利用一步水热合成法即可制得,并且无需繁琐的样品预处理和纯化过程。此外,所述碳点无需额外标记其他物质即可高选择性的检测喹诺酮类抗生素,只在碳点溶液中加入喹诺酮类抗生素,碳点的荧光发生明显淬灭。本发明荧光碳点可在免标记检测喹诺酮类抗生素中应用。
本发明提供的一种荧光碳点免标记检测喹诺酮类抗生素的方法,步骤为:
1)、配置浓度为0.2mg/mL的碳点溶液;
2)、配置浓度梯度为0.05、0.10、0.15……0.95、1.0mM的喹诺酮类抗生素的标准溶液;
3)、向碳点溶液中加入喹诺酮类抗生素,使碳点的荧光逐渐淬灭;
4)、测定碳点反应前后的荧光强度,根据喹诺酮类抗生素的浓度和相对荧光强度变化值之间的关系建立检测喹诺酮类的标准曲线;
5)、定量检测:测定待测样品和碳点反应前后的荧光强度变化,计算反应前后相对荧光强度变化值,参照步骤4)中获得的标准曲线,得到待测样品中喹诺酮类抗生素的含量。
以上所述喹诺酮类抗生素可以为诺氟沙星、环丙沙星、氧氟沙星或左氧氟沙星。
本发明具有以下有益技术效果:
(1)本发明提供的碳点原材料廉价易得,运用一步合成法,无需繁琐的预处理和纯化过程,制备过程节能省时。
(2)本发明的提供碳点在水溶液中都具有良好的溶解度和分散性;能发射明亮黄色荧光,在长时间光照与高离子强度下光学性质稳定。
(3)本发明所述的荧光碳点的表面良好地保留了原料化合物的化学结构,可与喹诺酮类抗生素直接发生相互作用,促使碳点的荧光淬灭,基于这项原理,所述碳点可作为荧光探针免标记检测喹诺酮类抗生素。
(4)本发明所述的荧光碳点在检测喹诺酮类抗生素的应用与传统的喹诺酮类抗生素检测方法相比,无需修饰额外的标记物,无需利用昂贵的分析仪器即可达到良好的检测效果,具有优异的选择性和灵敏度,在实际操作中更具有优越性。同时此项应用也拓展了碳点的应用范围。
附图说明
图1为实施例1制备的碳点的荧光发射光谱及紫外吸收光谱;
图2为实施例1制备的碳点的AFM图谱;
图3为实施例1制备的碳点的XPS图谱;
图4为实施例1制备的碳点检测诺氟沙星的荧光发射光谱图;
图5为碳点、碳点/诺氟沙星溶液体系在紫外灯下的照片;
图6为其他抗生素对碳点检测诺氟沙星体系无荧光干扰的柱状图。
具体实施方式
下面结合实施例对本发明做详细说明,实施例给出了详细的实施方式和具体的操作过程,但本发明的保护范围不限于下述的实施例。
实施例1
碳点的制备方法:
1)、分别取0.30g邻苯二胺和氨基丁酸置于玻璃烧杯中,加入20mL去离子水中,充分搅拌后超声溶解15分钟;
2)、将反应混合物转移至水热反应釜中,并置于烘箱内,160℃反应8h,得到棕色溶液;
3)、取出水热反应釜,自然冷却,将反应产物稀释10倍,在13000r/m转速下离心15min,去除不溶物得到澄清黄色的溶液,用0.22μm滤膜过滤后得到纯净的碳点的水溶液;
4)、将上述碳点水溶液冷冻干燥后得到碳点固体。
实施例2
将实施例1制备的荧光碳点进行荧光激发、发射和紫外吸收光谱表征(见图1),进行AFM和XPS表征(见图2-3),得到本发明制备的荧光碳点发射黄色荧光,为单分散无定型纳米颗粒,表面含有氨基、羧基、羟基基团。
实施例3
取实施例1制备的荧光碳点水溶液(0.2mg/mL)2mL置于荧光比色皿中,分别加入0.2mL浓度为0.05、0.10、0.15……0.95、1.0mM的诺氟沙星溶液,混合均匀,在荧光光度计中扫描发射光谱(λex=420nm,λem=564nm),根据诺氟沙星的浓度和相对荧光强度变化值之间的关系,计算碳点对诺氟沙星的检测范围及检出限。(见图4)
实施例4
如图5,将实施例1制备的荧光碳点水溶液置于玻璃瓶中,在365nm紫外灯下的图片,碳点溶液为黄色荧光(左),加入诺氟沙星后荧光明显淬灭。
实施例5
取实施例1制备的荧光碳点水溶液(0.2mg/mL)2.0mL置于荧光比色皿中,分别加入10μL浓度为10mM的诺氟沙星(NOR)、土霉素(OTC)、氨苄西林(AMP)、红霉素(ERY)、氯霉素(CHL)、硫酸链霉素(STR)溶液,混合均匀,在荧光光度计中扫描发射光谱(λex=420nm,λem=564nm),记录加入不同抗生素前后,碳点溶液的荧光强度,通过计算相对荧光强度观察碳点对喹诺酮类抗生素的选择性。见图6,与其他种类抗生素相比,只有喹诺酮类抗生素——诺氟沙星能使碳点的荧光淬灭。
Claims (6)
1.一种黄色荧光碳点的制备方法,其特征在于步骤为:
1)、将邻苯二胺和氨基丁酸置于玻璃烧杯中,加入去离子水中,充分搅拌后超声溶解15-20分钟,邻苯二胺和氨基丁酸的质量比为:0.15-0.62∶0.162-0.648;
2)、将反应混合物转移至水热反应釜中,并置于烘箱内,160℃反应8-10h,得到棕色溶液;
3)、取出水热反应釜,自然冷却,将反应产物加入去离子水稀释10-20倍,在13000r/m转速下离心15min,去除不溶物得到澄清棕黄色的溶液,用0.22μm滤膜过滤后得到纯净的碳点水溶液;
4)、将上述碳点水溶液冷冻干燥后得到黄色荧光碳点。
2.如权利要求1所述方法制备的黄色荧光碳点。
3.如权利要求2所述的荧光碳点在免标记检测喹诺酮类抗生素中的应用。
4.如权利要求3所述的荧光碳点在免标记检测喹诺酮类抗生素中的应用,所述的喹诺酮类抗生素为诺氟沙星、环丙沙星、氧氟沙星或左氧氟沙星。
5.一种荧光碳点免标记检测喹诺酮类抗生素的方法,其特征在于步骤为:
1)、配置浓度为0.2mg/mL的碳点溶液;
2)、配置浓度梯度为0.05、0.10、0.15……0.95、1.0mM的喹诺酮类抗生素的标准溶液;
3)、向碳点溶液中加入喹诺酮类抗生素,使碳点的荧光逐渐淬灭;
4)、测定碳点反应前后的荧光强度,根据喹诺酮类抗生素的浓度和相对荧光强度变化值之间的关系建立检测喹诺酮类的标准曲线;
5)、定量检测:测定待测样品和碳点反应前后的荧光强度变化,计算反应前后相对荧光强度变化值,参照步骤4)中获得的标准曲线,得到待测样品中喹诺酮类抗生素的含量。
6.如权利要求5所述的一种荧光碳点免标记检测喹诺酮类抗生素的方法,其特征在于所述的喹诺酮类抗生素为诺氟沙星、环丙沙星、氧氟沙星或左氧氟沙星。
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