CN116496290A - 一种基于吡喃-香豆素染料的肼荧光探针的制备和应用 - Google Patents
一种基于吡喃-香豆素染料的肼荧光探针的制备和应用 Download PDFInfo
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Abstract
本发明涉及了一种基于吡喃‑香豆素的肼(N2H4)荧光探针的制备和应用，该荧光探针的结构式为：本发明提供了以3‑乙酰基‑7‑羟基香豆素、4‑二乙氨基‑2‑羟基‑苯甲酰苯甲酸、乙酰丙酸等为原料合成该荧光探针的制备方法；首先，该荧光探针对N2H4表现出较高的灵敏度，探针与N2H4反应之后荧光显著增强；其次，该荧光探针对N2H4表现出很高的选择性，不受其他活性氧，活性硫，生物硫醇，氨基酸，有机胺的干扰；并且，该荧光探针与N2H4作用迅速，响应时间在15分钟以内；此外，该荧光探针可应用于实际水样中N2H4的检测。
Description
技术领域
本发明属于荧光探针技术领域，具体涉及一种基于吡喃-香豆素染料的肼荧光探针的制备和应用。
背景技术
肼(N2H4)是一种具有强还原性和亲核性的试剂，并广泛用于光敏剂、药物、农药、染料及其他化学品的制造(S.D.Zelnick,D.R.Mattie,P.C.Stepaniak,Aviat.Space.Environ.Med.,2003,74,1285-1291；U.Ragnarsson,Chem.Soc.Rev.,2001,30,205-213；K.Yamada,K.Yasuda,N.Fujiwara,Z.Siroma,H.Tanaka,Y.Miyazaki,T.Kobayashi,Electrochem.Commun.,2003,5,892-896)。肼可以广泛运用在不同领域，由于具有良好的水溶性，极容易通过皮肤和口腔被人体所吸收。作为一种神经毒素，若人体过量接触肼，可能导致肝、肺、肾和神经系统严重损伤(S.Garrod,M.E.Bollard,A.W.Nicholls,S.C.Connor,J.Connelly,J.K.Nicholson,E.Holmes,Chem.Res.Toxicol.,2005,18,115-122；K.Kucukoglu,H.I.Gul,P.Taslimi,I.Gulcin,C.T.Supuran,Bioorg.Chem.,2019,86,316-321)。据报道，N2H4已被美国环境保护署(EPA)列为可能致癌物质，并建议将10ppb列为阈值。因此，需要寻求一种既简便又高效的检测手段来检测肼。
荧光方法具有灵敏度高、操作简单和响应速度快等优点而显示出很大的应用潜力(Y.Q.Tan,J.C.Yu,J.K.Gao,Y.J.Cui,Y.Yang,G.D.Qian,Dyes Pigment,2013,99,966-971；L.Q.Yan,S.Q.Zhang,Y.Xie,X.Y.Mu，J.B.Zhu,Crit.Rev.Anal.Chem.,2022,52,210-229)。到目前为止，已经开发了一些检测肼的荧光探针，用于实时检测环境和细胞的浓度(X.Dai,Z.Y.Wang,Z.F.Du,J.Y.Miao,B.X.Zhao,Sens.Actuators B Chem.,2016,232,369-374；R.Chen,G.J.Shi,J.J.Wang,H.F.Qin,Q.Zhang,S.J.Chen,Y.G.Wen,J.B.Guo,K.P.Wang,Z.Q.Wang,Y.Wang,Spectrochim.Acta A Mol.Biomol.Spectrosc.,2021,252,119510；X.L.Xue,Q.Zhang,K.P.Wang,S.J.Chen,L.S.Tang,Z.Q.Hu,Spectrochim.Acta AMol.Biomol.Spectrosc.,2022,279,121406)。但是，这些探针存在一些问题：(1)荧光探针的灵敏度较差；(2)这些荧光探针具有相对较长的响应时间，反应时间在30分钟以上。因此，设计和合成具有灵敏度高和响应速度快的荧光探针是十分迫切的。
吡喃-香豆素染料是目前荧光探针领域中应用比较广泛的一类染料，它具有摩尔吸光系数大、荧光量子产率高等优势。据报道，利用吡喃-香豆素荧光探针已经检测了许多目标物，如：Cys,HClO,H2O2和Hg2+等(J.Liu,Y.Q.Sun,P.Wang,J.Zhang,W.Guo,Analyst,2013,138,2654-2660；H.Lv,X.F.Yang,Y.Zhong,Y.Guo,Z.Li,H.Li,Anal.Chem.,2014,86,1800-1807；S.Ding,Q.Zhang,S.Xue,G.Feng,Analyst,2015,140,4687-4693；B.Dong,X.Song,X.Kong,C.Wang,Y.Tang,Y.Liu,W.Lin,Adv.Mater.,2016,28,8755-8759)。但是，到现在为止，还没有以吡喃-香豆素为染料，乙酰丙酸为识别基团的荧光探针来灵敏且快速检测N2H4。因此，设计和合成一种基于吡喃-香豆素染料的荧光探针来检测N2H4是非常有必要的。
发明内容
根据所提出的要求，本发明人对此进行了深入研究，在付出了大量创造性劳动后，提供了一种基于吡喃-香豆素染料的肼荧光探针。
本发明的技术方案是，一种基于吡喃-香豆素染料的肼荧光探针，其结构式如下：
一种基于吡喃-香豆素染料的肼荧光探针的制备方法。步骤如下：
在100mL的圆底烧瓶中，将1当量的化合物CP-OH，3～5当量的乙酰丙酸、1.5当量的4-二甲氨基吡啶和1.5当量1-乙基-3-(3-二甲基氨基丙基)碳二亚胺溶解到15～25mL二氯甲烷中，反应混合物室温下搅拌8～12h，停止反应，通过减压蒸馏除去溶剂，粗产品用体积比为100/1～100/3的CH2Cl2/CH3OH作洗脱剂进行柱层析，得到紫色固体产物，即为所述的荧光探针CPN。
本发明的有益效果是，一种基于吡喃-香豆素染料的肼荧光探针的良好的光谱响应性能。首先，研究该探针的荧光光谱性质。探针本身在676nm处没有明显的荧光发射；加入N2H4后，在676nm处出现了明显的发射。并且，随着N2H4浓度的增大，探针近红外荧光不断增强。当加入20μM的N2H4时，荧光强度增强4倍。该探针的检测范围从2μM到20μM，检测限为0.67μM，这说明该探针可以高灵敏的检测N2H4。接着，研究探针的紫外吸收光谱。探针本身在620nm附近无吸收带，加入N2H4后，620nm附近出现新吸收峰。然后，研究探针的选择性。考察了探针与无机离子(Na+、Mg2+、K+、Cu2+、Zn2+、NH4 +、Cl-、Br-、I-、HCO3 -、NO3 -、SO4 2-)，活性氧(H2O2、ClO-)，活性硫(HS-、HSO3 -)，生物硫醇(Cys、GSH)，氨基酸(Leu、Thr、Trp、Lys、Phe、Met、Val、Ile)，有机胺(二甲胺、对硝基苯胺)以及检测物肼(N2H4)的荧光响应情况。结果发现，只有N2H4引起荧光光谱的改变，其他检测物对探针的荧光光谱没有明显的影响。最后，研究了pH值对荧光探针测定N2H4的影响，当pH值在7.0到8.0之间时，不影响荧光探针对N2H4的测定。此外，该荧光探针响应比较迅速，响应时间在15分钟以内。
一种基于吡喃-香豆素染料的肼荧光探针的应用。N2H4一般存在于水中，湖水和自来水被用来评估荧光探针在实际样品中的应用。为了确保实验的准确性，将取得的湖水和自来水样本先静置2小时，然后将两者pH调至7.4备用。接着，配制成含有不同浓度的N2H4样品，进行荧光测试，在湖水中获得了98.81％～106.35％的回收率，在自来水中获得了95.00％～106.75％的回收率。这些结果说明，探针能够检测实际样品中的N2H4，为监测水中的N2H4提供了一种可靠的手段。
附图说明
图1为荧光探针的合成路线。
图2为荧光探针与不同浓度的N2H4作用后的荧光光谱图。
横坐标为波长，纵坐标为荧光强度。荧光探针的浓度为10.0μM，N2H4的浓度分别为：0,2.0,4.0,6.0,8.0,10.0,12.0,14.0,16.0,18.0,20.0μM。发射波长范围为640-740nm，对应的激发波长为620nm。
图3为荧光探针对不同N2H4浓度的荧光线性响应图。
图4为荧光探针及荧光探针与N2H4作用后的紫外可见吸收光谱图。
横坐标为波长，纵坐标为吸光度。荧光探针的浓度为10.0μM，N2H4浓度为20.0μM。
图5为荧光探针的选择性图。
荧光探针的浓度为10.0μM，N2H4浓度为20.0μM，其它分析物浓度均为20.0μM。
图6为pH对荧光探针的影响图。
图7为荧光探针与N2H4作用后荧光强度随时间变化的曲线图，N2H4浓度为10.0,15.0,20.0μM。
具体实施方式
下面结合附图和具体实施例对本发明进行详细说明，但不限于此。
实施例1：
荧光探针的合成
合成路线如图1。在100mL的圆底烧瓶中，将1当量的化合物CP-OH，3当量的乙酰丙酸、1.5当量的4-二甲氨基吡啶和1.5当量的1-乙基-3-(3-二甲基氨基丙基)碳二亚胺溶解到25mL二氯甲烷中，反应混合物室温下搅拌2h，停止反应，通过减压蒸馏除去溶剂，粗产品用体积比为100/1～100/3的CH2Cl2/CH3OH作洗脱剂进行柱层析，得到紫色固体产物(产率为40％)，即为所述的荧光探针。1H NMR(400MHz,CDCl3,δ,ppm)δ7.96(d,J＝7.5Hz,1H),7.63(d,J＝7.5Hz,1H),7.24(s,2H),7.11(t,J＝3.9Hz,2H),6.76(s,2H),6.50(s,1H),6.31(s,2H),5.4(s,1H),3.34(q,J＝12.0Hz,4H),2.88–2.83(t,J＝8.0Hz,4H),2.23(s,3H),1.18(t,J＝8.0Hz,6H).13C NMR(101MHz,CDCl3,δ,ppm)δ206.80,171.28,170.20,158.05,154.32,154.17,153.87,153.01,149.83,146.26,139.82,135.00,131.39,130.15,129.91,128.97,126.98,125.50,124.41,119.27,118.65,117.01,110.18,109.71,105.18,104.11,97.74,44.90,38.30,30.14,28.08,13.00.MS(TOF):579.19
实施例2：
荧光探针和N2H4溶液配制
称取一定量荧光探针CPN固体溶解在DMSO中来制备1.0×10-3mol·L-1的CPN备用溶液。N2H4溶液的配制：将一定量的N2H4溶解在二次蒸馏水中，转移到10mL的容量瓶中，加水至刻度线，得到浓度为1.0×10-2mol·L-1的N2H4。将50μL CPN备用溶液，1.2mL DMSO和不同体积的N2H4备用溶液加入5mL的容量瓶，最终通过HEPES缓冲溶液来定容，得到浓度为1.0×10-5mol·L-1的荧光探针和2.0×10-6～2.0×10-5mol·L-1的N2H4混合待测溶液。
实施例3：
荧光探针与N2H4作用的荧光光谱的测定
图2为荧光探针与N2H4作用的荧光光谱，荧光探针的浓度为10μM，N2H4的浓度依次为0,2.0,4.0,6.0,8.0,10.0,12.0,14.0,16.0,18.0,20.0μM。实验所用激发波长为620nm，发射波长范围为640～740nm。狭缝宽度为10.0nm/10.0nm，所用的荧光测定仪器为日立F4600荧光分光光度计。从图2可以看出，加入N2H4之前，由于乙酰丙酯基团的淬灭作用，探针本身几乎没有发射峰；随着N2H4的加入，在676nm处发射峰增强。并且，随着N2H4浓度的增大，探针的荧光强度不断增强。图3为探针对不同N2H4浓度的线性响应图。荧光强度跟N2H4的浓度呈现线性关系，该探针的检测范围从2.0μM到20.0μM，检测限为0.67μM。这说明该探针可以高灵敏的检测N2H4。
实施例4：
荧光探针与N2H4作用的紫外可见吸收光谱的测定
图4为荧光探针与N2H4作用后的紫外可见吸收光谱图，荧光探针的浓度为10.0μM，N2H4的加入量为20.0μM。紫外可见吸收光谱测定用的仪器为安捷伦Cary60紫外可见分光光度计。从图4中可以看出，探针本身在620nm处无吸收带；加入N2H4之后，620nm处的出现新的吸收峰。
实施例5：
荧光探针对N2H4测定的选择性
图5为荧光探针对N2H4测定的选择性图。考察在浓度为10.0μM的荧光探针中加入N2H4(20.0μM)以及无机离子(Na+、Mg2+、K+、Cu2+、Zn2+、NH4 +、Cl-、Br-、I-、HCO3 -、NO3 -、SO4 2-)，活性氧(ClO-、H2O2)，氨基酸(Leu、Thr、Trp、Lys、Phe、Met、Val、Lle))，活性硫(HS-、HSO3 -)，生物硫醇(Cys、GSH)，有机胺(p-Nitroaniline、Dimethylamine)的荧光响应情况。从图5可以看出，只有N2H4能引起荧光光谱的明显增强，其他检测物对探针的荧光光谱没有明显的影响。这些结果表明，荧光探针对N2H4有良好的选择性。
实施例6：
溶液pH值对荧光探针测定N2H4的荧光性质的影响
考察pH值对荧光探针测定N2H4的荧光光谱的影响，其结果如图6。我们研究的pH范围为6.0-9.5，荧光探针的浓度为10.0μM，N2H4的浓度为20.0μM。从图中可以看出，荧光探针随着pH的变化，荧光强度基本不变，说明pH对探针本身没有影响。然而，加入N2H4之后，在pH在7.0～8.0范围内，荧光强度比值显著增强。综上所述，当pH值在7.0到8.0之间时，不影响荧光探针对N2H4的测定，是比较合适的pH值范围，这非常有利于该探针用于实际样品中N2H4的测定。
实施例7：
荧光探针与N2H4作用的响应时间的测定
我们研究了荧光探针对N2H4的响应时间，其结果如图7。从图中可以看出，该探针对N2H4的响应时间为15min，这能够满足在实际样品中进行监测的要求。从图7还可以看出，荧光强度达到最大值后，在之后的时间里，荧光强度不再发生变化，这表明此荧光探针光稳定性较好。
实施例8：
荧光探针应用于湖水样品的检测
湖水样本取自湘潭大学画眉潭，样品静置2小时，将静置后的湖水pH调至7.4。由于湖水样本不含有N2H4，因此需要外加N2H4配制成待测混合溶液，然后进行荧光检测，结果列于表1。从表中可以看出，探针对湖水中N2H4的回收率为98.81％～106.35％。
表1湖水中N2H4加标回收率的测定
实施例9：
荧光探针应用于自来水样品的检测
湖水样本取自湘潭大学化学化工大楼自来水水龙头，将自来水的pH调至7.4。由于自来水样本不含有N2H4，因此需要外加N2H4配制成待测混合溶液，然后进行荧光检测，结果列于表2。从表中可以看出，探针对自来水中N2H4的回收率为95.00％～106.75％。
表2自来水中N2H4加标回收率的测定
Claims (3)
1.一种基于吡喃-香豆素染料的肼荧光探针，即CPN，其特征在于，结构如下：
2.根据权利要求1所述的一种基于吡喃-香豆素染料的肼荧光探针的制备方法，其特征在于，反应步骤如下：
在100mL的圆底烧瓶中，将1当量的化合物CP-OH，3～5当量的乙酰丙酸、1.5当量的4-二甲氨基吡啶和1.5当量1-乙基-3-(3-二甲基氨基丙基)碳二亚胺溶解到15～25mL二氯甲烷中，反应混合物室温下搅拌1～2h，停止反应，通过减压蒸馏除去溶剂，粗产品用体积比为100/1～100/3的CH2Cl2/CH3OH作洗脱剂进行柱层析，得到紫色固体产物，即为所述的荧光探针CPN。
3.根据权利要求1所述的一种基于吡喃-香豆素染料的肼荧光探针的应用，其特征在于，所述荧光探针可应用于实际水样中肼含量的检测。
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Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20120016128A1 (en) * | 2010-07-16 | 2012-01-19 | Zhenjun Diwu | Rhodamine lactone phosphoramidites and polymers |
| WO2012074693A1 (en) * | 2010-11-16 | 2012-06-07 | Enzo Biochem, Inc. | Self-immolative probes for enzyme activity detection |
| KR20130077402A (ko) * | 2011-12-29 | 2013-07-09 | 중앙대학교 산학협력단 | 히드라진 선택성을 갖는 화학센서 및 이를 이용한 히드라진 검출방법 |
| CN105017196A (zh) * | 2015-07-21 | 2015-11-04 | 山东大学 | 一种检测肼的近红外比率荧光探针及其应用 |
| WO2016182444A1 (en) * | 2015-05-12 | 2016-11-17 | Rijksuniversiteit Groningen | 3d-printable antimicrobial composite resins, methods for manufacturing the same |
| CN109988560A (zh) * | 2019-05-17 | 2019-07-09 | 济南大学 | 一种新型香豆素衍生物的肼荧光探针 |
| CN111303102A (zh) * | 2019-11-29 | 2020-06-19 | 福建医科大学孟超肝胆医院(福州市传染病医院) | 一种硝基还原酶响应的乏氧探针化合物及其制备与应用 |
| CN114113045A (zh) * | 2020-08-31 | 2022-03-01 | 湖南超亟检测技术有限责任公司 | 一种新型近红外水合肼荧光检测试剂及其应用 |
| CN115215841A (zh) * | 2022-07-13 | 2022-10-21 | 河南理工大学 | 一种吡啶鎓-香豆素衍生物荧光探针及其制备方法和应用 |
-
2023
- 2023-03-24 CN CN202310305291.4A patent/CN116496290B/zh active Active
Patent Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20120016128A1 (en) * | 2010-07-16 | 2012-01-19 | Zhenjun Diwu | Rhodamine lactone phosphoramidites and polymers |
| WO2012074693A1 (en) * | 2010-11-16 | 2012-06-07 | Enzo Biochem, Inc. | Self-immolative probes for enzyme activity detection |
| KR20130077402A (ko) * | 2011-12-29 | 2013-07-09 | 중앙대학교 산학협력단 | 히드라진 선택성을 갖는 화학센서 및 이를 이용한 히드라진 검출방법 |
| WO2016182444A1 (en) * | 2015-05-12 | 2016-11-17 | Rijksuniversiteit Groningen | 3d-printable antimicrobial composite resins, methods for manufacturing the same |
| CN105017196A (zh) * | 2015-07-21 | 2015-11-04 | 山东大学 | 一种检测肼的近红外比率荧光探针及其应用 |
| CN109988560A (zh) * | 2019-05-17 | 2019-07-09 | 济南大学 | 一种新型香豆素衍生物的肼荧光探针 |
| CN111303102A (zh) * | 2019-11-29 | 2020-06-19 | 福建医科大学孟超肝胆医院(福州市传染病医院) | 一种硝基还原酶响应的乏氧探针化合物及其制备与应用 |
| CN114113045A (zh) * | 2020-08-31 | 2022-03-01 | 湖南超亟检测技术有限责任公司 | 一种新型近红外水合肼荧光检测试剂及其应用 |
| CN115215841A (zh) * | 2022-07-13 | 2022-10-21 | 河南理工大学 | 一种吡啶鎓-香豆素衍生物荧光探针及其制备方法和应用 |
Non-Patent Citations (4)
| Title |
|---|
| MIAOMIAO XING ET AL.: "A coumarin chalcone ratiometric fluorescent probe for hydrazine based on deprotection, addition and subsequent cyclization mechanism", 《CHEM. COMMUN.》, vol. 55, 20 November 2019 (2019-11-20), pages 14980 - 14983 * |
| MYUNG GIL CHOI ET AL.: "Hydrazine-Selective Chromogenic and Fluorogenic Probe Based on Levulinated Coumarin", 《ORG. LETT.》, vol. 13, no. 19, 14 September 2011 (2011-09-14), pages 5260 - 5263 * |
| ZUJUN XU ET AL.: "A new colorimetric and far‐red fluorescent probe for hydrazine with a large red‐shifted absorption spectrum", 《LUMINESCENCE》, vol. 32, 31 December 2017 (2017-12-31), pages 466 - 470 * |
| 刘维岩等: "反应型荧光探针在肼检测中的应用", 《化学试剂》, vol. 40, no. 6, 30 June 2018 (2018-06-30), pages 507 - 512 * |
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