Rapid Colorimetric Sensor Based on Gold Nanoparticles Functionalized 4-Amino-3-hydrazino-5-mercapto-1,2,4-triazole for Cortisol Detection in Saliva Sample


Hanim Istatik Badi'ah(1*), Ni Nyoman Tri Puspaningsih(2), Ganden Supriyanto(3), Nasronudin Nasronudin(4)

(1) Department of Chemistry, Faculty of Science and Technology, Airlangga University, Jl. Dr. Ir. H. Soekarno, Mulyorejo, Surabaya 60115, Indonesia; Department of Medical Laboratory Technology, Institute of Health Science Banyuwangi, Jl. Letkol Istiqlah No. 109, Banyuwangi 68422, Indonesia
(2) Department of Chemistry, Faculty of Science and Technology, Airlangga University, Jl. Dr. Ir. H. Soekarno, Mulyorejo, Surabaya 60115, Indonesia
(3) Department of Chemistry, Faculty of Science and Technology, Airlangga University, Jl. Dr. Ir. H. Soekarno, Mulyorejo, Surabaya 60115, Indonesia
(4) Department of Medicine, Faculty of Medicine, Airlangga University, Jl. Dr. Ir. H. Soekarno, Mulyorejo, Surabaya 60115, Indonesia
(*) Corresponding Author


The rapid, simple, and selective colorimetric sensing method of cortisol has been successfully developed using AuNPs modified with 4-amino-3-hydrazino-5-mercapto-1,2,4-triazole (AuNPs-AHMT). The principle of this method is based on the color change from wine red to purple (redshift) when AuNPs-AHMT interacts with cortisol. The hydrogen bonding between the hydroxyl group from cortisol and the amine group from AHMT induces the aggregation of AuNPs. The modification of the AuNPs surface with AHMT aims to increase its stability. The properties of AuNPs and AuNPs-AHMT were characterized by UV-vis spectrophotometer. The interaction between AuNPs-AHMT and cortisol was studied by UV-vis and FTIR spectroscopies. The proposed method was optimized and validated. Au(III) was reduced to AuNPs at an optimum NaBH4 concentration of 1.0 mM. Validation of the proposed method showed good analytical performance with linearity from 1.0–50.0 nM, accuracy 91.07–102.77%, intra-day precision < 2.22% and inter-day precision < 2.17%, detection limit 0.76 nM, quantification limit 2.54 nM, and sensitivity 0.0112 nM/mL. The proposed method also showed good selectivity with the presence of some interferences in the sample. The proposed method was successfully applied for the determination of cortisol in the saliva by the standard addition method with acceptable recovery.


AHMT; colorimetric sensing; cortisol; gold nanoparticles

Full Text:

Full Text PDF


[1] Mohd Azmi, N.A.S., Juliana, N., Azmani, S., Mohd Effendy, N., Abu, I.F., Mohd Fahmi Teng, N.I., and Das, S., 2021, Cortisol on circadian rhythm and its effect on the cardiovascular system, Int. J. Environ. Res. Public Health, 18 (2), 676.

[2] Ramamoorthy, S., and Cidlowski, J.A., 2016, Corticosteroids: Mechanisms of action in health and disease, Rheum. Dis. Clin. N. Am., 42 (1), 15–31.

[3] Dahlgren, A., Kecklund, G., Theorell, T., and Åkerstedt, T., 2009, Day-to-day variation in saliva cortisol–Relation with sleep, stress and self-rated health, Biol. Psychol., 82 (2), 149–155.

[4] Hakamata, Y., Komi, S., Moriguchi, Y., Izawa, S., Motomura, Y., Sato, E., Mizukami, S., Kim, Y., Hanakawa, T., Inoue, Y., and Tagaya, H., 2017, Amygdala-centred functional connectivity affects daily cortisol concentrations: A putative link with anxiety, Sci. Rep., 7 (1), 8313.

[5] Lee, D.Y., Kim, E., and Choi, M.H., 2015, Technical and clinical aspects of cortisol as a biochemical marker of chronic stress, BMB Rep., 48 (4), 209–216.

[6] Barugh, A.J., Gray, P., Shenkin, S.D., MacLullich, A.M., and Mead, G.E., 2014, Cortisol levels and the severity and outcomes of acute stroke: A systematic review, J. Neurol., 261 (3), 533–545.

[7] Aguilar Cordero, C.M., Sánchez López, A.M., Mur Villar, N., García García, I., Rodríguez López, M.A., Ortegón Piñero, A., and Cortés Castell, E., 2014, Cortisol salival como indicador de estrés fisiológico en niños y adultos; Revisión sistemática, Nutr. Hosp., 29 (5), 960–968.

[8] Perogamvros, I., Keevil, B.G., Ray, D.W., and Trainer, P.J., 2010, Salivary cortisone is a potential biomarker for serum free cortisol, J. Clin. Endocrinol. Metab., 95 (11), 4951–4958.

[9] Estrada-Y-Martin, R.M., and Orlander, P.R., 2011, Salivary cortisol can replace free serum cortisol measurements in patients with septic shock, Chest, 140 (5), 1216–1222.

[10] Boolani, A., Channaveerappa, D., Dupree, E.J., Jayathirtha, M., Aslebagh, R., Grobe, S., Wilkinson, T., and Darie, C.C., 2019, “Trends in Analysis of Cortisol and Its Derivatives” in Advances in Experimental Medicine and Biology, Eds. Woods, Alisa G., and Darie, C.C., Springer International Publishing, Cham, Switzerland, 649–664.

[11] Pappachan, J.M., Hariman, C., Edavalath, M., Waldron, J., and Hanna, F.W., 2017, Cushing’s syndrome: A practical approach to diagnosis and differential diagnoses, J. Clin. Pathol., 70 (4), 350–359.

[12] Russell, E., Kirschbaum, C., Laudenslager, M.L., Stalder, T., de Rijke, Y., van Rossum, E.F., Van Uum, S., and Koren, G., 2015, Toward standardization of hair cortisol measurement: Results of the first international interlaboratory round robin, Ther. Drug Monit., 37 (1), 71–75.

[13] Kannankeril, J., Carroll, T., Findling, J.W., Javorsky, B., Gunsolus, I.L., Phillips, J., and Raff, H., 2020, Prospective evaluation of late-night salivary cortisol and cortisone by EIA and LC-MS/MS in suspected cushing syndrome, J. Endocr. Soc., 4 (10), bvaa107.

[14] Raff, H., and Phillips, J.M., 2019, Bedtime salivary cortisol and cortisone by LC-MS/MS in healthy adult subjects: Evaluation of sampling time, J. Endocr. Soc., 3 (8), 1631–1640.

[15] Yang, X., Yu, Y., and Gao, Z.A., 2014, Highly sensitive plasmonic DNA assay based on triangular silver nanoprism etching, ACS Nano, 8, 4902–4907.

[16] Ma, X., Chen, Z., Kannan, P., Lin, Z., Qiu, B., and Guo, L., 2016, Gold nanorods as colorful chromogenic substrates for semiquantitative detection of nucleic acids, proteins, and small molecules with the naked eye, Anal. Chem., 88 (6), 3227–3234.

[17] Zhang, Z., Wang, H., Chen, Z., Wang, X., Choo, J., and Chen, L., 2018, Plasmonic colorimetric sensors based on etching and growth of noble metal nanoparticles: Strategies and applications, Biosens. Bioelectron., 114, 52–65.

[18] Zhou, W., Gao, X., Liu, D., and Chen, X., 2015, Gold nanoparticles for in vitro diagnostics, Chem. Rev., 115 (19), 10575–10636.

[19] Cabuzu, D., Cirja, A., Puiu, R., and Grumezescu, A.M., 2015, Biomedical applications of gold nanoparticles, Curr. Top. Med. Chem., 15 (16), 1605–1613.

[20] Tvrdonova, M., Vlcnovska, M., Vanickova, L.P., Kanicky, V., Adam, V., Ascher, L., Jakubowski, N., Vaculovicova, M., and Vaculovic, T., 2019, Gold nanoparticles as labels for immunochemical analysis using laser ablation inductively coupled plasma mass spectrometry, Anal. Bioanal. Chem., 411 (3), 559–564.

[21] Sadiq, Z., Safiabadi Tali, S.H., Hajimiri, H., Al-Kassawneh, M., and Jahanshahi-Anbuhi, S., 2023, Gold nanoparticles-based colorimetric assays for environmental monitoring and food safety evaluation, Crit. Rev. Anal. Chem., 1–36.

[22] Chang, K., Wang, S., Zhang, H., Guo, Q., Hu, X., Lin, Z., Sun, H., Jiang, M., and Hu, J., 2017, Colorimetric detection of melamine in milk by using gold nanoparticles-based LSPR via optical fibers, PLoS One, 12 (5), e0177131.

[23] Badi'ah, H.I., Seedeh, F., Supriyanto, G., and Zaidan, A.H., 2019, Synthesis of silver nanoparticles and the development in analysis method, IOP Conf. Ser.: Earth Environ. Sci., 217, 012005.

[24] Mauriz, E., 2020, Clinical applications of visual plasmonic colorimetric sensing, Sensors, 20 (21), 6214.

[25] Saha, K., Agasti, S.S., Kim, C., Li, X., and Rotello, V.M., 2012, Gold nanoparticles in chemical and biological sensing, Chem. Rev., 112 (5), 2739–2779.

[26] Bolaños, K., Kogan, M.J., and Araya, E., 2019, Capping gold nanoparticles with albumin to improve their biomedical properties, Int. J. Nanomed., 14, 6387–6406.

[27] Slavgorodska, M.V., Gurova, Y.O., and Kyrychenko, A., 2021, γ-Cyclodextrin as a capping agent for gold nanoparticles, Comput. Theor. Chem., 1194, 113060.

[28] Li, F., He, T., Wu, S., Peng, Z., Qiu, P., and Tang, X., 2021, Visual and colorimetric detection of uric acid in human serum and urine using chitosan stabilized gold nanoparticles, Microchem. J., 164, 105987.

[29] Thambiliyagodage, C., 2022, Ligand exchange reactions and PEG stabilization of gold nanoparticles, Curr. Res. Green Sustainable Chem., 5, 100245.

[30] Bouduban, M.E.F., Burgos-Caminal, A., Ossola, R., Teuscher, J., and Moser, J.E., 2017, Energy and charge transfer cascade in methylammonium lead bromide perovskite nanoparticle aggregates, Chem. Sci., 8 (6), 4371–4380.

[31] Chan, K.L., Fawcett, D., and Poinern, G.E.J., 2016, Gold nanoparticle treated textile-based materials for potential use as wearable sensors, Int. J. Sci., 5 (5), 82–89.

[32] De Souza, C.D., Nogueria, B.R., and Rostelato, M.E.C.M., 2019, Review of the methodologies used in the synthesis gold nanoparticles by chemical reduction, J. Alloys Compd., 798, 714–740.

[33] Kesik, M., Kanik, F.E., Hizalan, G., Kozanoglu, D., Esenturk, E.N., Timur, S., and Toppare, L., 2013, A functional immobilization matrix based on a conducting polymer and functionalized gold nanoparticles: Synthesis and its application an amperometric glucose biosensor, Polymer, 54 (17), 4463–4471.

[34] Mehravani, B., Ribeiro, A.I., and Zille, A., 2021, Gold nanoparticles synthesis and antimicrobial effect on fibrous materials, Nanomaterials, 11 (5), 1067.

[35] Radich, J.G., and Kamat, P.V., 2013, Making graphene holey gold nanoparticle mediated hydroxyl radical attack on reduced graphene oxide, ACS Nano, 7, 5546–5557.

[36] Oliveira, J.P., Prado, A.R., Keijok, W.J., Ribeiro, M.R.N., Pontes, M.J., Nogueira, B.V., and Guimarães, M.C.C., 2020, A helpful method for controlled synthesis of monodisperse gold nanoparticles through response surface modeling, Arabian J. Chem., 13 (1), 216–226.

[37] Khezri, S., Bahram, M., and Samadi, N., 2018, Hydrogen bonding recognition and colorimetric detection of isoprenaline using 2-amino-5-mercapto-1,3,4-thiadiazol functionalized gold nanoparticles, Spectrochim. Acta, Part A, 189, 522–527.

[38] Qin, L., Zeng, G., Lai, C., Huang, D., Zhang, C., Xu, P., Hu, T., Liu, X., Cheng, M., Liu, Y., Hu, L., and Zhou, Y., 2017, A visual application of gold nanoparticles: Simple, reliable and sensitive detection of kanamycin based on hydrogen bonding, Sens. Actuators, B, 243, 946–954.

DOI: https://doi.org/10.22146/ijc.80874

Article Metrics

Abstract views : 1261 | views : 587

Copyright (c) 2023 Indonesian Journal of Chemistry

Creative Commons License
This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.


Indonesian Journal of Chemistry (ISSN 1411-9420 /e-ISSN 2460-1578) - Chemistry Department, Universitas Gadjah Mada, Indonesia.

Analytics View The Statistics of Indones. J. Chem.