CN108587611A - 一种双波长荧光金纳米簇的合成方法及应用 - Google Patents
一种双波长荧光金纳米簇的合成方法及应用 Download PDFInfo
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Abstract
本发明公开了一种双波长荧光金纳米簇的合成方法,本发明以富含多糖的石斛为碳量子原料,通过一步热分解法合成荧光碳量子点,采用碳量子点作为合成荧光金纳米簇的还原剂和保护剂,合成双波长荧光金纳米簇;在360nm的激发波长下,荧光金纳米簇同时在蓝光区(443nm)和红光区(714nm)产生发射光谱,以荧光金纳米簇作为温度及酪氨酸的荧光探针,构建了比率荧光传感器;443nm的蓝光区对温度变化具有线性响应,温度由5℃升至75℃过程中,荧光强度逐渐减弱,温度下降时,荧光恢复;而酪氨酸对714nm红光区具有线性增敏效应,荧光强度随酪氨酸浓度的增大而增大,从而建立了温度及酪氨酸的比率荧光探针;建立的方法具有强的特异性及高的灵敏度。
Description
技术领域
本发明涉及化学分析检测技术领域,具体为一种双波长荧光金纳米簇的合成方法及应用。
背景技术
金纳米簇,由几十至几百个金原子构成,是一种近年来引起科学家极大研究兴趣的荧光纳米材料,其粒径通常小于2nm。由于其粒径接近电子的费米波长,因此其具有与大粒径的金纳米颗粒不同的光学性质、化学性质和电学性质。作为一种新型荧光纳米材料,金纳米簇具有合成简单、光稳定性强、生物兼容性好等优点,目前已经成为纳米领域里发展前景最好的材料之一, 在生命医学以及生物传感领域中被广泛应用。在荧光传感模式中,比率型荧光传感器利用两个或多个发射峰强度的比值变化带来输出颜色的改变,可以实现对目标分析物的特异、准确检测。与单一发射的荧光传感器相比,比率传感器通过建立内标,极大削弱了探针浓度、温度、溶剂极性、激发强度、环境的pH值等众多难以控制因素的干扰,使得结果更加精准,响应范围更宽。
金纳米簇合成中热还原法将三价Au离子还原为零价的Au原子,进而形成荧光金纳米簇,但容易聚集形成粒径较大且没有荧光的金纳米颗粒。为了解决这个问题,一些配体分子被修饰到金纳米簇表面以避免其发生聚集进而增强其稳定性。选择合适的表面修饰模板分子对于金纳米簇的成功制备起到了至关重要的作用。碳量子点,如同石墨烯量子点及碳纳米点,是一类碳的新型碳材料。它具有优异的光学性能,可调的激发和发射行为,较高的荧光稳定性,较低的毒性和良好的生物相容性,在越来越多的领域中得到了广泛的应用。碳量子点表面丰富的羟基,使其成为金纳米簇合成中表面修饰很好的还原剂及稳定剂。目前,用碳量子点作为还原剂及稳定剂合成金纳米簇的方法,未见报道,更未见合成材料用于两种荧光比率探针的报道。
发明内容
本发明的目的在于提供一种以碳量子点作为还原剂和保护剂合成双波长荧光金纳米簇的方法,及其用于高灵敏、高选择性的温度及酪氨酸的荧光探针。
本发明双波长荧光金纳米簇合成方法包括以下步骤:
(1)氮掺杂碳量子点的合成:称取2-5g干燥的铁皮石斛粉末,分散于90-150mL 60-70℃的热水中并不断搅拌,待搅拌均匀呈糊状后再加入10-20mL无水乙醇和1-3mL乙二胺,搅拌均匀后,转移至聚四氟乙烯内衬反应釜中于180℃加热9-12h,自然冷却后,离心,除去大颗粒杂质,上清液过0.22μm滤膜后得到氮掺杂的碳量子点,将氮掺杂的碳量子点置于60-65℃下真空干燥24h,备用;
(2)以氮掺杂的碳量子点作为还原剂和保护剂合成荧光金纳米簇:取步骤(1)的氮掺杂的碳量子点,配制成浓度为3-5mg/mL水溶液,搅拌下油浴加热至90-120℃,加入浓度为0.2mg/mL的氯金酸四水合物溶液100-150μL,继续加热搅拌;待溶液由黄色逐渐变为红棕色后停止加热搅拌,所得产物即为双波长荧光金纳米簇。
所述步骤(1)中离心是在转速8000-10000rpm下进行15-20min。
本发明另一目的是提供上述方法制得的双波长荧光金纳米簇,双波长荧光金纳米簇的最大激发波长为365nm,最大发射波长分别为433nm及714nm;且在443nm波长处温度变化对荧光强度有线性响应,在714nm波长处酪氨酸浓度变化对荧光强度有线性增敏效应。
本发明另一目的是将上述方法制得的双波长荧光金纳米簇应用在作为温度及酪氨酸含量测定的荧光探针中,具体步骤如下:
(1)利用443nm波长对温度变化对荧光强度有线性响应,而在714nm波长无响应,在水温由5℃升至75℃过程中,加入双波长荧光金纳米簇,在365nm波长的光照激发下,获得433nm和714nm波长处荧光强度的比值I433/I714与温度的线性关系;
(2)利用714nm波长处酪氨酸浓度变化对荧光强度有线性增敏效应,而对443nm波长无响应,在浓度为0.25-125μmol/L的酪氨酸溶液中,加入双波长荧光金纳米簇,在365nm波长的光照激发下, 获得714nm和433nm波长处荧光强度的比值I714/I433与酪氨酸的线性关系,用于定量检测酪氨酸的浓度;
(3)在待测样品中加入双波长荧光金纳米簇,在365nm波长的光照激发下,测定待测样品的温度及酪氨酸的相应的比率荧光,并代入步骤(1)、步骤(2)的线性关系中,获得待测样品的温度及酪氨酸含量。
所述步骤(1)中双波长荧光金纳米簇的用量为50-200μL。
所述步骤(2)中双波长荧光金纳米簇的用量100-300 μL。
本发明的优点在于:
1、本发明采用以富含多糖的石斛为碳量子原料,通过一步热分解法合成荧光碳量子点,采用碳量子点作为合成荧光金纳米簇的还原剂和保护剂,合成荧光金纳米簇,合成的荧光金纳米簇具有蓝光区(443nm)和红光区(714nm)的两个发射波长;
2、石斛为原料合成的碳量子荧光量子产率25%;荧光金纳米簇在水溶液中分散均匀且稳定,纳米粒径约为7nm,对后续应用奠定了基础;
3、温度及酪氨酸选择性在两个发射波长处分别有荧光响应,在两个波长处荧光强度的比值与分析物浓度成线性关系,温度可识别5℃至75℃,酪氨酸检测限可以达到0.05μM,其他氨基酸没有干扰,方法具有较好的特异性;
本发明以富含多糖的石斛为碳量子原料,通过一步热分解法合成荧光碳量子点,采用碳量子点作为合成荧光金纳米簇的还原剂和保护剂,合成荧光金纳米簇。在360nm的激发波长下,金纳米簇同时在蓝光区(443nm)和红光区(714nm)产生发射光谱,以荧光金纳米簇作为温度及酪氨酸的荧光探针,构建了比率荧光传感器,443nm的蓝光区对温度变化具有线性响应,温度由5℃升至75℃过程中,荧光强度逐渐减弱,温度下降时,荧光恢复;而酪氨酸对714nm红光区具有线性增敏效应,荧光强度随酪氨酸浓度的增大而增大;该方法新颖,特异性强,灵敏度高。
附图说明
图1为实施例1中双波长荧光金纳米簇的扫描电镜图;
图2为实施例1中双波长荧光金纳米簇的激发发射光谱示意图谱;
图3为温度的比率荧光探针以相对荧光强度比值I433/I714对温度示意图谱;
图4为酪氨酸的比率荧光探针以相对荧光强度比值I714/I433对酪氨酸浓度示意图谱。
具体实施方式
下面将结合具体的实施例对本发明的技术方案作进一步详细地描述说明,但本发明的保护范围并不仅限于此。
实施例1:血液中温度及色氨酸的含量测定操作步骤如下:
(1)碳量子点的合成:称取2g干燥的铁皮石斛粉末,分散于90mL 60℃的热水中并不断搅拌,待搅拌均匀呈糊状后再加入10mL无水乙醇和1mL乙二胺,搅拌均匀后,转移至聚四氟乙烯内衬反应釜于180℃加热9h,自然冷却后,8000rpm下离心20min,除去大颗粒杂质,上清液过0.22μm滤膜后得到氮掺杂的碳量子点,以硫酸奎宁为标准物,其荧光量子产率25%;将所得量子点置于60℃真空干燥箱中干燥24h,备用;
(2)以碳量子点作为还原剂和保护剂合成荧光金纳米簇:量取步骤(1)的氮掺杂的碳量子点,配制成浓度为3mg/mL的水溶液置于10mL圆底烧瓶中,搅拌下油浴加热至95℃,加入浓度0.2 mg/mL的氯金酸四水合物溶液100μL,继续加热搅拌;待溶液由黄色逐渐变为红棕色后停止加热搅拌,所得产物即为双波长荧光金纳米簇,由图1可知纳米粒径约为7nm;
(3)荧光金纳米簇的激发波长及发射波长确定:将合成的荧光金纳米簇溶解于水溶液中,在300到800nm进行扫描,荧光金纳米簇的最大激发波长在365nm,最大发射波长分别在433nm及714nm处(图2);
(4)比率探针对温度的荧光响应:在水的温度由5℃升至75℃过程中,加入双波长荧光金纳米簇100μL,在荧光光谱365nm波长的光照激发下,记录波段从300到800nm;探针以相对荧光强度比值I433/I714对温度进行作图,得到温度在5℃-75℃线性关系I433/I714=0.01258+0.0809T(℃) (r= 0.9985),用于温度的测定;图3为温度的比率荧光探针以相对荧光强度比值I433/I714对温度示意图谱,说明有好的线性关系;
(5)比率探针对酪氨酸的荧光响应:在浓度为0.25-125μmol/L的酪氨酸溶液中,加入双波长荧光金纳米簇200μL,在荧光光谱365nm波长的光照激发下,记录波段从300到800nm;探针以相对荧光强度比值I714/I433对酪氨酸浓度进行作图,得到的线性关系I714/I433=0.2752+0.0087C(r= 0.9971),检出限为 0.05μM ( S /N = 3),用于定量检测酪氨酸的浓度;图4为酪氨酸的比率荧光探针以相对荧光强度比值I714/I433对酪氨酸浓度示意图谱,说明有好的线性关系;
(6)为了验证本方法对酪氨酸检测的选择性,选择了一系列常见干扰物质进行实验;结果表明,与酪氨酸的响应相比,亮氨酸、半胱氨酸、胱氨酸、天冬氨酸、色氨酸、壳聚糖、丝氨酸、精氨酸、葡萄糖及vitamin C 和对本探针的响应较小,说明本方法对酪氨酸的检测具有优良的选择性;
(7)血液样品测定:取空腹静脉血2mL置于肝素抗凝管内,于4℃、10000rpm离心15min分离血浆,取肝素抗凝血浆 200μL,加入20μL质量浓度35%的高氯酸溶液,涡漩混匀1 min,静置10 min,于4℃、15000rpm离心力下离心10min 沉淀血浆中的蛋白质,取上清液按步骤(3)及步骤(4)进行温度及酪氨酸比率荧光测定,代入步骤(4)、步骤(5)的线性关系中,得到血液温度为32℃,色氨酸含量为7.81μmol/L。
实施例2:发酵液样品中温度及酪氨酸的含量测定步骤为:
(1)碳量子点的合成:称取3g干燥的铁皮石斛粉末,分散于110mL 65℃的热水中并不断搅拌,待搅拌均匀呈糊状后再加入15mL无水乙醇和2mL乙二胺,搅拌均匀后,转移至聚四氟乙烯内衬反应釜中于180℃加热10h,自然冷却后,离心,除去大颗粒杂质,上清液过0.22μm滤膜后得到氮掺杂的碳量子点,将氮掺杂的碳量子点置于65℃下真空干燥24h,备用;
(2)以氮掺杂的碳量子点作为还原剂和保护剂合成荧光金纳米簇:取步骤(1)的氮掺杂的碳量子点,配制成浓度为4mg/mL水溶液置于10mL圆底烧瓶中,搅拌下油浴加热至110℃,加入浓度为0.2mg/mL的氯金酸四水合物溶液120μL,继续加热搅拌;待溶液由黄色逐渐变为红棕色后停止加热搅拌,所得产物即为双波长荧光金纳米簇。
(3)比率探针对温度的荧光响应:同实施例1步骤(4);
(4)比率探针对酪氨酸的荧光响应:同实施例1步骤(5);
(5) 样品测定:采用酪氨酸合成酶酶促发酵转化合成酪氨酸,发酵液稀释100倍后,按实施例1步骤(4)及(5)进行温度及酪氨酸比率荧光测定,其中测定温度加入步骤(1)合成的荧光金纳米簇200μL,测定酪氨酸加入步骤(1)合成的荧光金纳米簇300μL,代入步骤(3)、步骤(4)的线性关系中,得到发酵液温度为27℃,酪氨酸含量为125.50μM。
实施例3:牛奶样品中温度及酪氨酸含量测定步骤为:
(1)碳量子点的合成:称取5g干燥的铁皮石斛粉末,分散于150mL 70℃的热水中并不断搅拌,待搅拌均匀呈糊状后再加入20mL无水乙醇和3mL乙二胺,搅拌均匀后,转移至聚四氟乙烯内衬反应釜中于180℃加热11h,自然冷却后,离心,除去大颗粒杂质,上清液过0.22μm滤膜后得到氮掺杂的碳量子点,将氮掺杂的碳量子点置于62℃下真空干燥24h,备用;
(2)以氮掺杂的碳量子点作为还原剂和保护剂合成荧光金纳米簇:取步骤(1)的氮掺杂的碳量子点,配制成浓度为5mg/mL水溶液置于10mL圆底烧瓶中,搅拌下油浴加热至120℃,加入浓度为0.2mg/mL的氯金酸四水合物溶液150μL,继续加热搅拌;待溶液由黄色逐渐变为红棕色后停止加热搅拌,所得产物即为双波长荧光金纳米簇。
(3)比率探针对温度的荧光响应:同实施例1步骤(4);
(4)比率探针对酪氨酸的荧光响应:同实施例1步骤(5);
(5) 样品测定:准确称取脱脂奶0.1g于有刻度10 mL具塞试管中,加入4.00 mL木瓜蛋白酶溶液,振荡,置65 ℃恒温箱中酶解过夜,次日取出酶解液冷却,以3000r/min离心 15min,吸取上清液1 mL,按实施例1步骤(4)及(5)进行温度及酪氨酸比率荧光测定,其中测定温度加入步骤(1)合成的荧光金纳米簇200μL,测定酪氨酸加入步骤(1)合成的荧光金纳米簇300μL,代入步骤(3)、步骤(4)的线性关系中,得到牛奶样品温度为34℃,酪氨酸含量为43.91μM。
Claims (4)
1.一种双波长荧光金纳米簇的合成方法,其特征在于,包括以下步骤:
(1)氮掺杂碳量子点的合成:称取2-5g干燥的铁皮石斛粉末,分散于90-150mL 60-70℃的热水中并不断搅拌,待搅拌均匀呈糊状后再加入10-20mL无水乙醇和1-3mL乙二胺,搅拌均匀后,转移至聚四氟乙烯内衬反应釜中于180℃加热9-12h,自然冷却后,离心,除去大颗粒杂质,上清液过0.22μm滤膜后得到氮掺杂的碳量子点,将氮掺杂的碳量子点置于60-65℃下真空干燥24h,备用;
(2)以氮掺杂的碳量子点作为还原剂和保护剂合成荧光金纳米簇:取步骤(1)的氮掺杂的碳量子点,配制成浓度为3-5mg/mL水溶液,搅拌下油浴加热至90-120℃,加入浓度为0.2mg/mL的氯金酸四水合物溶液100-150μL,继续加热搅拌;待溶液由黄色逐渐变为红棕色后停止加热搅拌,所得产物即为双波长荧光金纳米簇。
2.根据权利要求书1所述的双波长荧光金纳米簇的合成方法,其特征在于:步骤(1)中离心是在转速8000-10000rpm下进行15-20min。
3.权利要求书1所述的合成方法制得的双波长荧光金纳米簇,其特征在于:双波长荧光金纳米簇的最大激发波长为365nm,最大发射波长分别为433nm及714nm;且在443nm波长处温度变化对荧光强度有线性响应,在714nm波长处酪氨酸浓度变化对荧光强度有线性增敏效应。
4.权利要求3所述的双波长荧光金纳米簇在作为温度及酪氨酸含量测定的荧光探针中的应用,其特征在于,具体步骤如下:
(1)利用443nm波长对温度变化对荧光强度有线性响应,而在714nm波长无响应,在水温由5℃升至75℃过程中,加入双波长荧光金纳米簇,在365nm波长的光照激发下,获得433nm和714nm波长处荧光强度的比值I433/I714与温度的线性关系;
(2)利用714nm波长处酪氨酸浓度变化对荧光强度有线性增敏效应,而对443nm波长无响应,在浓度为0.25-125μmol/L的酪氨酸溶液中,加入双波长荧光金纳米簇,在365nm波长的光照激发下, 获得714nm和433nm波长处荧光强度的比值I714/I433与酪氨酸的线性关系,用于定量检测酪氨酸的浓度;
(3)在待测样品中加入双波长荧光金纳米簇,在365nm波长的光照激发下,测定待测样品的温度及酪氨酸的相应的比率荧光,并代入步骤(1)、步骤(2)的线性关系中,获得待测样品的温度及酪氨酸含量。
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CN111961466A (zh) * | 2020-07-27 | 2020-11-20 | 太原理工大学 | 一种用于检测肝素的碳量子点荧光探针 |
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Citations (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN105067577A (zh) * | 2015-07-14 | 2015-11-18 | 天津大学 | 一种可视化检测汞离子的碳点-金纳米团簇双发射比率型荧光探针及制备方法 |
CN106047342A (zh) * | 2016-06-23 | 2016-10-26 | 南京理工大学 | 一种镉离子和抗坏血酸检测用的碳量子点/金团簇比率荧光探针 |
CN106141200A (zh) * | 2015-03-26 | 2016-11-23 | 上海交通大学 | 一种碳点/金复合纳米粒子的制备方法及用途 |
CN106908427A (zh) * | 2017-03-01 | 2017-06-30 | 哈尔滨师范大学 | 金纳米簇和碳量子点复合荧光探针及其应用 |
CN107543809A (zh) * | 2017-07-28 | 2018-01-05 | 郑州大学 | 一种谷胱甘肽分子检测用试纸条及检测方法 |
-
2018
- 2018-05-10 CN CN201810440618.8A patent/CN108587611B/zh active Active
Patent Citations (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN106141200A (zh) * | 2015-03-26 | 2016-11-23 | 上海交通大学 | 一种碳点/金复合纳米粒子的制备方法及用途 |
CN105067577A (zh) * | 2015-07-14 | 2015-11-18 | 天津大学 | 一种可视化检测汞离子的碳点-金纳米团簇双发射比率型荧光探针及制备方法 |
CN106047342A (zh) * | 2016-06-23 | 2016-10-26 | 南京理工大学 | 一种镉离子和抗坏血酸检测用的碳量子点/金团簇比率荧光探针 |
CN106908427A (zh) * | 2017-03-01 | 2017-06-30 | 哈尔滨师范大学 | 金纳米簇和碳量子点复合荧光探针及其应用 |
CN107543809A (zh) * | 2017-07-28 | 2018-01-05 | 郑州大学 | 一种谷胱甘肽分子检测用试纸条及检测方法 |
Non-Patent Citations (2)
Title |
---|
CHUANXI WANG等: "Tunable Carbon-Dot-Based Dual-Emission Fluorescent Nanohybrids for Ratiometric Optical Thermometry in Living Cells", 《ACS APPLIED MATERIALS & INTERFACES》 * |
EEPSITA PRIYADARSHINI等: "Au@carbon dot nanoconjugates as a dual mode enzyme-free sensing platform for cholesterol", 《JOURNAL OF MATERIALS CHEMISTRY B》 * |
Cited By (7)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN111961466A (zh) * | 2020-07-27 | 2020-11-20 | 太原理工大学 | 一种用于检测肝素的碳量子点荧光探针 |
CN114136957A (zh) * | 2021-11-16 | 2022-03-04 | 中国中医科学院中药研究所 | 一种免仪器的石斛产地可视化溯源方法 |
CN114136957B (zh) * | 2021-11-16 | 2024-01-23 | 中国中医科学院中药研究所 | 一种免仪器的石斛产地可视化溯源方法 |
CN115006345A (zh) * | 2022-06-28 | 2022-09-06 | 上海交通大学医学院附属瑞金医院 | 一种口服零价钼纳米点的制备方法与应用 |
CN115006345B (zh) * | 2022-06-28 | 2023-11-28 | 上海交通大学医学院附属瑞金医院 | 一种口服零价钼纳米点的制备方法与应用 |
CN117110256A (zh) * | 2023-05-29 | 2023-11-24 | 兰州大学第一医院 | 一种基于N-GQDs荧光淬灭原理的尿液酪氨酸检测试剂及检测方法 |
CN117110256B (zh) * | 2023-05-29 | 2024-04-19 | 兰州大学第一医院 | 一种基于N-GQDs荧光淬灭原理的尿液酪氨酸检测试剂及检测方法 |
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