一种用于硫化氢检测的荧光开启型探针的合成及应用

doi:10.6023/cjoc202603030

摘要/Abstract

硫化氢(H₂S)不仅是一种有毒的环境污染物(常见于食品腐败及工业废气中), 也是生物体内重要的信号分子. 因此, 对其进行精准检测具有重要的理论与实际意义. 本研究通过4-(二苯氨基)-3-羟基-[1,1-联苯]-4-甲醛(TPA-CHO)与2-氨基苯硫酚反应, 合成了具有高荧光量子产率(Φ=35.6%)的荧光团TPAB-OH. 进一步通过其与2,4-二硝基氟苯反应, 制备了一种新型HS^-荧光探针TPAB-HS. 该探针表现出优异的检测性能, 包括宽的pH适用范围(5~12)、短的响应时间(10 min)、高的信噪比以及低的检测限(8.561×10^-7mol/L). 此外, TPAB-HS探针成功应用于人乳腺癌细胞(MCF-7)中外源性HS^-的荧光成像, 以及实际水样和食品样品中HS^-的检测, 展示了其在生物成像和复杂样品分析领域的应用潜力.

关键词: 三苯胺衍生物, 荧光探针, 硫化氢, 食品样品, 荧光成像

Hydrogen sulfide (H₂S) is not only a toxic environmental pollutant (commonly found in food spoilage and industrial waste gas) but also an important signaling molecule in organisms. Therefore, it is necessary to develop an effective method to detect hydrogen sulfide. In this paper, TPAB-OH with high fluorescence quantum yield (Φ=35.6%) was designed and synthesized by combining 4-(diphenylamino)-3-hydroxy-[1,1-biphenyl]-4-formaldehyde (TPA-CHO) with 2-aminobenzene-thiol. TPAB-OH reacted with 2,4-dinitrofluorobenzene to produce a new probe TPAB-HS, which has the advantages of a wide pH range (5~12), fast response (10 min), high signal-to-noise ratio and low detection limit (8.56×10^-7 mol/L). In addition, the probe TPAB-HS is also suitable for the detection of HS^- in human breast cancer cells (MCF-7), actual water samples and food samples.

Key words: triphenylamine derivatives, fluorescence probe, hydrogen sulfide, food samples, fluorescence imaging

引用此文

王龙, 蒙亚男, 任睿祺, 田明玉, 汤立军. 一种用于硫化氢检测的荧光开启型探针的合成及应用[J]. 有机化学, 2026, 46(10): 3963-3969.

Long Wang, Yanan Meng, Ruiqi Ren, Mingyu Tian, Lijun Tang. Synthesis and Application of a Fluorescence Turn-on Probe for Hydrogen Sulfide Detection[J]. Chinese Journal of Organic Chemistry, 2026, 46(10): 3963-3969.

导出引用 EndNote|Reference Manager|ProCite|BibTeX|RefWorks

分享此文

图/表 12

图式1. 探针TPAB-HS的合成路线

Scheme 1. Synthesis route of probe TPAB-HS

图1. 在不同浓度的HS-(0~350 μmol/L)存在下, TPAB-HS (10 μmol/L)在溶液THF/H2O (V/V=4/6)中的荧光光谱变化

Figure 1. Fluorescence spectra changes of TPAB-HS (10 μmol/ L) in THF/H2O (V/V=4/6) solution in the presence of HS- at different concentrations (0~350 μmol/L)

图2. 探针TPAB-HS的荧光强度(506 nm)与HS-浓度(0~350 μmol/L)之间的线性关系

Figure 2. Linear relationship between the fluorescence intensity of TPAB-HS (at 506 nm) and HS- concentration (0~350 μmol/ L)

图3. TPAB-HS (10 μmol/L)在THF/H2O (V/V=4/6)中加入各种分析物(350 μmol/L)后荧光光谱变化

Figure 3. Fluorescence spectral changes of TPAB-HS (10 μmol/L) in THF/H2O (V/V=4/6) upon addition of various analytes (350 μmol/L)

图4. 在加入各种阴离子以及生物硫醇(350 μmol/L)的TPAB-HS (10 μmol/L) THF/H2O (V/V=4/6)溶液中, 继续加入HS-前后的荧光强度变化

Figure 4. Fluorescence intensity changes of TPAB-HS (10 μmol/L) in THF/H2O (V/V=4/6) solution containing various anions and biothiols (350 μmol/L) before and after the addition of HS-

图5. 探针TPAB-HS和TPAB-HS＋HS-在不同pH条件下的荧光强度变化

Figure 5. Changes in the fluorescence intensity of the probe TPAB-HS and TPAB-HS＋HS- under different pH conditions

图6. 探针TPAB-HS (10 μmol/L)＋350 μmol/L H2S的荧光强度随时间的变化

Figure 6. Fluorescence intensity changes of the probe TPAB-HS (10 μmol/L) over time in the presence of 350 μmol/L H2S

图式2. 探针TPAB-HS响应HS-的机理

Scheme 2. Response mechanism of the probe TPAB-HS to HS-

图7. (a)探针TPAB-HS与HS-反应后分离产物的核磁共振氢谱; (b)化合物TPAB-OH的核磁共振氢谱; (c)探针TPAB-HS的核磁共振氢谱

Figure 7. (a) 1H NMR spectrum of the isolated product from the reaction of the probe TPAB-HS with HS-; (b) 1H NMR spectrum of compound TPAB-OH; (c) 1H NMR spectrum of the probe TPAB-HS

Sample	Added/ (μmol•L^-1)	Detect/ (μmol•L^-1)	Recovery/%	RSD/%
River water	40	38.32	95.81	1.11
	60	60.90	101.50	0.85
	80	85.86	107.32	2.97
Lake water	80	81.16	101.45	2.53
	100	97.35	97.35	0.78
	120	126.31	105.26	0.19
Tap water	40	41.26	103.16	1.42
	60	57.70	96.17	0.41
	80	78.54	98.17	0.83

表1. 探针TPAB-HS在实际水样中对HS-的检测a

Table 1. Detection of HS- by TPAB-HS in real water samples

Sample	Added/ (μmol•L^-1)	Detect/ (μmol•L^-1)	Recovery/%	RSD/%
Red Wine2	20	19.86	99.31	1.24
	40	41.13	102.86	0.88
	60	59.11	98.52	1.85
Red Wine1	20	22.14	110.69	1.26
	40	39.41	98.53	0.46
	60	57.95	96.59	1.62
Beer2	20	18.35	91.75	1.70
	30	30.64	102.13	1.46
	40	41.95	104.87	1.08
Beer1	80	77.59	96.99	3.16
	100	104.02	104.02	0.47
	120	117.83	98.19	0.31

表2. 探针TPAB-HS在食品样品中对HS-的检测a

Table 2. Detection of HS- by TPAB-HS in food samples

图8. MCF-7细胞与探针TPAB-HS (10 μmol/L)孵育30 min后, 加入HS- (0, 50, 100, 200, 500 μmol/L)继续孵育30 min的荧光图像(比例尺: 20 μm)

Figure 8. Fluorescence images of MCF-7 cells incubated with probe TPAB-HS (10 μmol/L) for 30 min and then incubated with HS- (0, 50, 100, 200, 500 μmol/L) for 30 min (scale bar: 20 μm)

参考文献 32

[1]	Yang Y. X.; Chen L.; Hu X. L.; Zhong K. L.; Li S. D.; Yan X. M.; Zhang J. L.; Tang L. J. Chin. J. Org. Chem. 2023, 43, 308 (in Chinese).
	(杨雅馨, 陈琳, 胡晓玲, 钟克利, 李世迪, 燕小梅, 张璟琳, 汤立军, 有机化学, 2023, 43, 308.) doi: 10.6023/cjoc202207011
[2]	Shen W.; Wang P.; Xie Z.; Zhou H.; Hu Y.; Fu M.; Zhu Q. Talanta 2021, 234, 122621. doi: 10.1016/j.talanta.2021.122621
[3]	Módis K.; Ju Y.; Ahmad A.; Untereiner A. A.; Altaany Z.; Wu L.; Szabo C.; Wang R. Pharmacol. Res. 2016, 113, 116. doi: 10.1016/j.phrs.2016.08.023
[4]	Wu T.; Zheng Z.; Ren J.; Yang Z.; Zhang N.; Guo K.; Chen D.; Gao M.; Chen Y.; Liang X.; Wang Y.; Chen W.; Su Y.; He J.; Wang F. Stem Cell Res. Ther. 2025, 16, 603. doi: 10.1186/s13287-025-04704-9
[5]	Fiorucci S.; Antonelli E.; Mencarelli A.; Orlandi S.; Renga B.; Rizzo G.; Distrutti E.; Shah V.; Morelli A. Hepatology 2005, 42, 539. pmid: 16108046
[6]	Kamoun P.; Belardinelli M.; Chabli A.; Lallouchi K.; Chadefaux-Vekemans B. Am. J. Med. Genet. A 2003, 116A, 310. doi: 10.1002/ajmg.a.v116a:3
[7]	Kamoun P. Med. Hypotheses 2001, 57, 389. pmid: 11516234
[8]	Yang B.; Su M.-M.; Xue Y.-S.; He Z.-X.; Xu C.; Zhu H.-L. Sens. Actuators, B Chem. 2018, 276, 456. doi: 10.1016/j.snb.2018.08.138
[9]	Kang J.; Huo F.; Yao Y.; Yin C. Dyes Pigm. 2019, 171, 107755. doi: 10.1016/j.dyepig.2019.107755
[10]	Lu Y.; Dong B.; Song W.; Kong X.; Mehmood A. H.; Lin W. Anal. Methods 2019, 11, 3301. doi: 10.1039/C9AY00927B
[11]	Zhao Q.; Huo F.; Zhang Y.; Wen Y.; Yin C. Spectrochim. Acta A Mol. Biomol. Spectrosc. 2019, 215, 297. doi: 10.1016/j.saa.2019.02.096 pmid: 30844677
[12]	Liu Y.; Ding Y.; Huang J.; Zhang X.; Fang T.; Zhang Y.; Zheng X.; Yang X. Dyes Pigm. 2017, 138, 112. doi: 10.1016/j.dyepig.2016.11.038
[13]	Lin X.; Lu X.; Zhou J.; Ren H.; Dong X.; Zhao W.; Chen Z. Spectrochim. Acta A Mol. Biomol. Spectrosc. 2019, 213, 416. doi: 10.1016/j.saa.2019.01.085
[14]	Luo Y.; Zhu C.; Du D.; Lin Y. Anal. Chim. Acta 2019, 1061, 1. doi: 10.1016/j.aca.2019.02.045
[15]	Renault K.; Sabot C.; Renard P. Eur. J. Org. Chem. 2015, 2015, 7992. doi: 10.1002/ejoc.v2015.36
[16]	Ma J.; Li F.; Li Q.; Li Y.; Yan C.; Lu X.; Guo Y. New J. Chem. 2018, 42, 19272. doi: 10.1039/C8NJ04208J
[17]	Ren M.; Deng B.; Kong X.; Zhou K.; Liu K.; Xu G.; Lin W. Chem. Commun. 2016, 52, 6415. doi: 10.1039/C6CC00966B
[18]	Sun L.; Wu Y.; Chen J.; Zhong J.; Zeng F.; Wu S. Theranostics 2019, 9, 77. doi: 10.7150/thno.30080
[19]	Takano Y.; Echizen H.; Hanaoka K. Antioxid. Redox Signal. 2017, 27, 669. doi: 10.1089/ars.2017.7070
[20]	Qi F.; Zhang Y.; Wang B.; Chen W.; Yang L.; Yang Z.; Song X. Sens. Actuators B Chem. 2019, 296, 126533. doi: 10.1016/j.snb.2019.05.010
[21]	Pan Y.; Li Y.; Sun X.; Tang L.; Yan X. Dyes Pigm. 2023, 210, 110985. doi: 10.1016/j.dyepig.2022.110985
[22]	Zhong K.; Li Y.; Hu X.; Li Y.; Tang L.; Sun X.; Li X.; Zhang J.; Meng Y.; Ma R.; Wang S.; Li J. Food Chem. 2024, 438, 137987. doi: 10.1016/j.foodchem.2023.137987
[23]	Zhong K.; He Y.; Deng L.; Yan X.; Li X.; Tang Y.; Hou S.; Tang L. Anal. Chim. Acta 2020, 1127, 49. doi: S0003-2670(20)30699-1 pmid: 32800137
[24]	Zhong K.; Hu X.; Zhou S.; Liu X.; Gao X.; Tang L.; Yan X. J. Agric. Food Chem. 2021, 69, 4628. doi: 10.1021/acs.jafc.1c00862
[25]	Zhang K.; Zhang J.; Xi Z.; Li L.-Y.; Gu X.; Zhang Q.-Z.; Yi L. Chem. Sci. 2017, 8, 2776. doi: 10.1039/c6sc05646f pmid: 28553513
[26]	Men J.; Yang X.; Zhang H.; Zhou J. Dyes Pigm. 2018, 153, 206. doi: 10.1016/j.dyepig.2017.12.036
[27]	Zhao Q.; Yin C.; Kang J.; Wen Y.; Huo F. Dyes Pigm. 2018, 159, 166. doi: 10.1016/j.dyepig.2018.06.029
[28]	Zhong K.; Chen L.; Pan Y.; Yan X.; Hou S.; Tang Y.; Gao X.; Li J.; Tang L. Spectrochim. Acta A Mol. Biomol. Spectrosc. 2019, 221, 117135. doi: 10.1016/j.saa.2019.117135
[29]	Zhong K.; Zhou S.; Yan X.; Li X.; Hou S.; Cheng L.; Gao X.; Li Y.; Tang L. Dyes Pigm. 2020, 174, 108049. doi: 10.1016/j.dyepig.2019.108049
[30]	Meng Y.; Yao X.; Zhong K.; Li Y.; Tang L. Eur. J. Org. Chem. 2023, 26, e202300022.
[31]	Jin L.; Tan X.; Dai L.; Bai H.; Wang Q. Spectrochim. Acta A Mol. Biomol. Spectrosc. 2019, 221, 117187. doi: 10.1016/j.saa.2019.117187
[32]	Kang J.; Huo F.; Yin C. Dyes Pigm. 2017, 146, 287. doi: 10.1016/j.dyepig.2017.07.016