A New Flow Injection System with Merging-Zone Technique for the Determination of Copper(II) by Neocuproine Reagent in Aqueous Solution


Ahmed Saleh Farhood(1*), Dakhil Nassir Taha(2)

(1) Department of Chemistry, College of Science, University of Babylon, Hilla 51002, Iraq
(2) Department of Chemistry, College of Sciences for Women, University of Babylon, Hilla 51002, Iraq
(*) Corresponding Author


A fast, simple, and high throughput sample merging-zone flow injection design was developed to determine copper(II) in aqueous solution. The procedure is based on the reduction of copper(II) to copper(I) by uric acid followed by a direct reaction with Neocuproine reagent (NC). The orange-yellow complex that forms absorb light at 454 nm. All conditions of the new flow injection unit were investigated. The analytical curve of copper(II) was linear with (r2) value of 0.9978, in the range of 0.4 to 40 mg/L with a detection limit of 0.1 mg/L and a quantification limit of 0.3 mg/L. the molar absorptivity was 1.661 × 105 L/mol cm and the recovery range was between 104.9 and 97%. The homemade acrylic valve was low-cost with zero dead volume and high repeatability (n = 7) with an RSD of 2.31%. The dispersion coefficient values were 1.8,1.62, and 1.31 for the concentrations of 5, 15, and 25 mg/L, respectively. The sample throughput was 69 h–1.


merging-zone; copper(II); Neocuproine; homemade valve; dead volume

Full Text:

Full Text PDF


[1] Abdel-Raouf, M.S., and Abdul-Raheim, A.R., 2017, Removal of heavy metals from industrial waste water by biomass-based materials: A review, J. Pollut. Eff. Control, 5 (1), 1000180.

[2] Ghorpade, A., and Ahammed, M.M., 2018, Water treatment sludge for removal of heavy metals from electroplating wastewater, Environ. Eng. Res., 23 (1), 92–98.

[3] Yu, J.S., Kim, S.H., Man, M.T., and Lee, H.S., 2018, Synthesis and characterization of water soluble fluorescent copper nanoparticles, Appl. Sci. Convergence Technol., 27 (4), 75–77.

[4] Jiang, T., Xie, D., Wu, J., He, H., Wang, H., Wang, N., Zhu, Z., Wang, Y., and Yang, T., 2020, Association between serum copper levels and prevalence of hyperuricemia: A cross-sectional study, Sci. Rep., 10 (1), 8687.

[5] Krivorotova, T., and Sereikaite, J., 2014, Determination of fructan exohydrolase activity in the crude extracts of plants, Electron. J. Biotechnol., 17 (6), 329–333.

[6] Dontha, S., 2016, A review on antioxidant methods, Asian J. Pharm. Clin. Res., 9 (8), 14–32.

[7] Zhang, G.Q., Li, Y.S., and Gao, X.F., 2018, An asynchronous-alternating merging-zone flow-injection gold nanoparticles probe method for determining anti-diabetic pioglitazone hydrochloride medicine, New J. Chem., 42 (6), 4337–4343.

[8] Proskurnin, M.A., Chernysh, V.V., Pakhomova, S.V., Kononets, M.Y., and Sheshenev, A.A., 2002, Investigation of the reaction of copper(I) with 2,9-dimethyl-1,10-phenanthroline at trace level by thermal lensing, Talanta, 57 (5), 831–839.

[9] Gaál, A., Garay, T.M., Horváth, I., Máthé, D., Szöllősi, D, Veres, D.S., Mbuotidem, J., Kovács, T., Tóvári, J., Bergmann, R., Streli, C., Szakács, G., Mihály, J., Varga, Z., and Szoboszlai, N., 2020, Development and in vivo application of a water-soluble anticancer copper ionophore system using a temperature-sensitive liposome formulation, Pharmaceutics, 12 (5), 466.

[10] Emir, G., Dilgin, Y., and Apak, R., 2020, A new redox mediator (cupric-neocuproine complex)-modified pencil graphite electrode for the electrocatalytic oxidation of H2O2: A flow injection amperometric sensor, ChemElectroChem, 7 (3), 649–658.

[11] Koga, T., Sakata, Y., and Terasaki, N., 2019, Accumulation and analysis of cuprous ions in a copper sulfate plating, J. Visualized Exp., 145, e59376.

[12] Skrovankova, S., Mlcek, J., Sochor, J., Baron, M., Kynicky, J., and Jurikova, T., 2015, Determination of ascorbic acid by electrochemical techniques and other methods, Int. J. Electrochem. Sci., 10, 2421–2431.

[13] Morosanova, M.A., and Morosanova, E.I., 2017, Silica-titania xerogel doped with Mo,P-heteropoly compounds for solid phase spectrophotometric determination of ascorbic acid in fruit juices, pharmaceuticals, and synthetic urine, Chem. Cent. J., 11 (1), 3.

[14] Losev, V.N., Didukh, S.L., Trofimchuk, A.K., and Zaporozhets, O.A., 2014, Adsorption–photometric and test determination of copper using silica gel sequentially modified with polyhexamethylene guanidine and bathocuproinedisulphonic acid, Adsorpt. Sci. Technol., 32 (6), 443–452.

[15] Ayaz, S., Dilgin, Y., and and Apak, R., 2020, Flow injection amperometric determination of hydrazine at a cupric-Neocuproine complex/anionic surfactant modified disposable electrode, Microchem. J., 159, 105457.

[16] Ribeiro, J.P.N., Magalhães, L.M., Reis, S., Lima, J.L.F.C., and Segundo, M.A., 2011, High-throughput total cupric ion reducing antioxidant capacity of biological samples determined using flow injection analysis and microplate-based methods, Anal. Sci., 27 (5), 483–488.

[17] Brasil, M.A.S., and Reis, B.F., 2017, An automated multicommuted flow analysis procedure for photometric determination of reducing sugars in wine employing a directly heated flow‑batch device, J. Braz. Chem. Soc., 28 (10), 2013–2020.

[18] Kukoc-Modun, L., Tsikas, D., Biocic, M., and Radić, N., 2015, Flow injection analysis of N-acetyl-L-cysteine based on the reduction of copper(II)-neocuproine reagent, Anal. Lett., 49 (5), 607–617.

[19] Chandramouleeswaran, S., and Ramkumar, J., 2018, Insight of spectrophotometric determination using 4-(2-pyridylazo)resorcinol: Application of stop flow injection analysis, Chem. Sin., 9 (2), 605–608.

[20] Kraljević, T., Jelić-Knezović, N., Marković Boras, M., and Ćurlin, M., 2020, Spectrophotometric hybrid flow system for determination of N-acetyl-L-cysteine in pharmaceuticals, IOSR J. Appl. Chem., 13 (5), 27–34.

[21] Omarova, S., Demir, S., and Andac, M., 2018, Development of a new spectrophotometric based flow injection analysis method for the determination of copper (II), J. Taibah Univ. Sci., 12 (6), 820–825.

[22] Segundo, M.A., Tóth, I.V., Magalhães, L.M., and Reis, S., 2015, “Automatic flow injection analysis (FIA) determination of total reducing capacity in serum and urine samples” in Advanced Protocols in Oxidative Stress III, Armstrong, D., Eds., Humana Press, New York, 277–284.‏

[23] Marques, S.S., Magalhães, L.M., Tóth, I.V., and Segundo, M.A., 2014, Insights on antioxidant assays for biological samples based on the reduction of copper complexes—the importance of analytical conditions, Int. J. Mol. Sci., 15 (7), 11387–11402.

[24] da Silva, P.A.B., de Souza, G.C.S., Paim, A.P.S., and Lavorante, A.F., 2018, Spectrophotometric determination of reducing sugar in wines employing in-line dialysis and a multicommuted flow analysis approach, J. Chil. Chem. Soc., 63 (2), 3994–4000.

[25] Viganor, L., Howe, O., McCarron, P., McCann, M., and Devereux, M., 2017, The antibacterial activity of metal complexes containing 1,10-phenanthroline: Potential as alternative therapeutics in the era of antibiotic resistance, Curr. Top. Med. Chem., 17 (11), 1280–1302.

[26] da Silva, J.C., Suarez, W.T., and de Oliveira Krambeck F.M., 2018, Flow-injection spectrophotometric determination of methimazole in pharmaceuticals using a charge-transfer complex Cu(I)‒neocuproine, J. Anal. Chem., 73 (3), 243–248.

[27] Taha, D.N., and Obaid, Z.S. 2016, Designing flow injection unit for chromates determining, Res. J. Pharm., Biol. Chem. Sci., 7 (6), 2242-2251.

[28] Babayeva, K., Demir, S., and Andac, M., 2017, A novel spectrophotometric method for the determination of copper ion by using a salophen ligand, N,N-disalicylidene-2,3-diaminopyridine, J. Taibah Univ. Sci., 11 (5), 808–814.

[29] Çağlar, Y., and Saka, E.T., 2017, Ionic liquid based dispersive liquideliquid microextraction procedure for the spectrophotometric determination of copper using 3-dimethylamino rhodanine as a chelating agent in natural waters, Karbala Int. J. Mod. Sci., 3 (4), 185–190.

[30] Kulkarni, A.A., and Vaidya, I.S., 2015, Flow injection analysis: An overview, J. Crit. Rev., 2 (4), 19–24.

[31] Yaseen, S.M., Qassim, B.B., and Al-Lami, N.O., 2020, Spectrophotometric determination of Co(II) in vitamin B12 using 2-(biphenyl-4-yl)-3-((2-(2,4-dinitrophenyl) hydrazono)methyl) imidazo [1,2-a]pyridine as ligand by flow injection–merging zone analysis, Al-Nahrain J. Sci., 23 (3), 24–38.

[32] Farhood, A.S., Majeed, A.S., Ali, L.A.M., and Taha, D.N., 2017, Semi-automated flow injection method for the determination of iron (II) by 1,10-phenanethroline, Orient. J. Chem., 33 (6), 3112–3120.

[33] Farhood, A.S., Ali, L.A.M., and Ali, F.F., 2017, Determination of aniline blue dye by flow injection analysis with home made valve, Orient. J. Chem., 33 (2), 944–950.

[34] Majeed, A.S., Farhood, A.S., Ali, L.A.M., and Taha, D.N., 2017, Home-made micro valve for determining malachite green dye by flow injection analysis, Indones. J. Chem., 17 (2), 248–255.

[35] Cassella, R.J., Magalhães, O.I., Couto, M.T., Lima, E.L.S., Neves, M.A.F.S., and Coutinho, F.M.B., 2005, Synthesis and application of a functionalized resin for flow injection/F AAS copper determination in waters, Talanta, 67 (1), 121–128.

[36] Mashhadizadeh, M.H., Pesteh, M., Talakesh, M., Sheikhshoaie, I., Ardakani, M.M., and Karimi, M.A., 2008, Solid-phase extraction of copper (II) by sorption on octadecyl silica membrane disk modified with a new Schiff base and determination with atomic absorption spectrometry, Spectrochim. Acta, Part B, 63 (8), 885–888.

[37] Şahin, Ç.A., and Tokgöz, İ., 2010, A novel solidified floating organic drop microextraction method for preconcentration and determination of copper ions by flow injection flame atomic absorption spectrometry, Anal. Chim. Acta, 667 (1-2), 83–87.

[38] Mohadesi, A., and Taher, M.A., 2007, Voltammetric determination of Cu(II) in natural waters and human hair at a meso-2,3-dimercaptosuccinic acid self-assembled gold electrode, Talanta, 72 (1), 95–100.

[39] Wainwright, P., Wadey, D., and Cook, P., 2018, An inductively coupled plasma mass spectrometry method for relative free copper determination and generation of a paediatric reference interval, Ann. Clin. Biochem., 55 (4), 485–490.

DOI: https://doi.org/10.22146/ijc.70799

Article Metrics

Abstract views : 2185 | views : 1360

Copyright (c) 2022 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.