Chemometric Optimization of Online SPE-LC Method Using Polypyrrole-Graphene Oxide Sorbent for Tetracycline Analysis in Water Samples
DOI:
https://doi.org/10.48048/tis.2026.10976Keywords:
Polypyrrole-graphene oxide, Online solid-phase extraction, Response surface methodology, Tetracycline antibiotics, Water samplesAbstract
A novel automated analytical method integrating online solid-phase extraction with liquid chromatography (online-SPE-LC) with chemometrically optimization was developed for the rapid and sensitive quantification of five selected tetracycline antibiotics, namely oxytetracycline, tetracycline, demeclocycline, chlortetracycline and doxycycline in water samples. In this method, a green polypyrrole-graphene oxide (PPy-GO) composite was employed as extraction sorbent. The effects of flow rate, valve switching time, sorbent mass, solvent composition and buffer pH were investigated using half-fraction Central Composite Design (CCD) of Response Surface Methodology (RSM). Optimal conditions were determined to be a solvent flow rate of 0.75 mL min⁻¹, valve switching time of 1.6 min, sorbent mass of 55 mg, solvent composition of 80:20 (ACN:water), and buffer pH of 2.5. The optimized method exhibited excellent linearity (R2 = 0.9990-0.9997) across a wide concentration range (10-1000 µg L−1) with low limits of detection (3.2 - 6.7 µg L−1). Method precision and accuracy were demonstrated by recoveries ranging from 82% - 102% and relative standard deviations (RSDs) of 0.6% - 3.0% (n=3) in both river and tap water samples. Compared to existing approaches, the developed method shows superior selectivity, high sensitivity, rapid and automation. Thus, this study highlights the great potential of PPy-GO as efficient sorbent integrated with online -SPE-LC method for the extraction of tetracycline from water samples.
HIGHLIGHTS
- A novel study using chemometrically Optimized Fully Automated Online-SPE-LC Method with Polypyrrole- Graphene Oxide as Sorbent toward tetracyclines antibiotics in water samples.
- Synthesis and Characterization of PPy-GO sorbent.
- Using Chemometric (BBD) optimization with fully automated online SPE-LC for tetracycline analysis in water samples using PPy-GO sorbent.
- The integrated system by Online SPE coupled with Liquid Chromatography as rapid developing technique for the analysis of tetracyclines antibiotics in water samples.
- Modern trend of extraction techniques for pharmaceutical analysis.
GRAPHICAL ABSTRACT
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F Granados-Chinchilla and C Rodriguez. Tetracyclines in food and feeding stuffs: From regulation to analytical methods, bacterial resistance, and environmental and health implications. Journal of Analytical Methods in Chemistry 2017; 2017(1), 1315497.
F Ahmad, D Zhu and J Sun. Environmental fate of tetracycline antibiotics: Degradation pathway mechanisms, challenges, and perspectives. Environmental Sciences Europe 2021; 33, 64.
SI Polianciuc, AE Gurzau, B Kiss, MG Stefan and F Loghin. Antibiotics in the environment: Causes and consequences. Medicine and Pharmacy Report 2020; 93(3), 231-240.
DG Larsson and CF Flach. Antibiotic resistance in the environment. Nature Reviews Microbiology 2022; 20, 257-269.
Y Amangelsin, Y Semenova, M Dadar, M Aljofan and G Bjorklund. The impact of tetracycline pollution on the aquatic environment and removal strategies. Antibiotics 2023; 12(3), 440.
D More, N Khan, RK Tekade and P Sengupta. An update on current trend in sample preparation automation in bioanalysis: Strategies, challenges and future direction. Critical Reviews in Analytical Chemistry 2024; 1, 1-25.
N Zaini, NSM Hanapi, N Saim, WNW Ibrahim and AL Anis. Selective determination of acidic drugs in water samples using online solid phase extraction liquid chromatography with alginate incorporated multi-walled carbon nanotubes as extraction sorbent. Indonesian Journal of Chemistry 2020; 20(5), 987-999.
ME Badawy, MA El-Nouby, PK Kimani, LW Lim and EI Rabea. A review of the modern principles and applications of solid-phase extraction techniques in chromatographic analysis. Analytical Sciences 2022; 38, 1457-1487.
I Reinholds, M Jansons, I Pugajeva and V Bartkevics. Recent applications of carbonaceous nanosorbents in solid phase extraction for the determination of pesticides in food samples. Critical Reviews in Analytical Chemistry 2019; 49(5), 439-458.
N Manousi, E Rosenberg, E Deliyanni, GA Zachariadis and V Samanidou. Magnetic solid-phase extraction of organic compounds based on graphene oxide nanocomposites. Molecules 2020, 25(5), 1148.
JJ Guo, AM Fiore, LT Murray, DA Jaffe, JL Schnell, CT Moore and GP Milly. Average versus high surface ozone levels over the continental USA: Model bias, background influences, and interannual variability. Atmospheric Chemistry and Physics 2018, 18(16), 12123-12140.
A Kaynak, A Zolfagharian, T Featherby, M Bodaghi, MP Mahmud and AZ Kouzani. Electrothermal modeling and analysis of Polypyrrole-coated wearable E-textiles. Materials 2021; 14(3), 550.
S Gao, Z Liu, Q Yan, P Wei, Y Li, J Ji and L Li. Facile synthesis of polypyrrole/reduced graphene oxide composite hydrogel for Cr(VI) removal. Journal of Inorganic and Organometallic Polymers and Materials 2021; 31, 3677-3685.
SNA Baharin, KP Sambasevam, NF Suhaimi, A Rahat, S Rahman and SN Musiran. Sunlight driven photocatalytic degradation of 2-chlorophenol by polypyrrole/graphene oxide composites. Malaysian Journal of Science 2021, 40(1), 67-79.
S Noreen, M Tahira, M Ghamkhar, I Hafiz, HN Bhatti, R Nadeem, MA Murtaza, M Yaseen, AA Sheikh, Z Naseem and F Younas. Treatment of textile wastewater containing acid dye using novel polymeric graphene oxide nanocomposites (GO/PAN, GO/PPy, GO/PSty). Journal of Materials Research and Technology 2021; 14, 25-35.
M Wang, X Duan, Y Xu and X Duan. Functional three-dimensional graphene/polymer composites. ACS Nano 2016; 10(8), 7231-7247.
S Sharma, RK Dubey and R Choudhary. Thermogram-based breast cancer detection using fractional derivative Sobel filtering and hybrid feature set in IoT framework. Computational Modeling in Engineering & Sciences 2022; 130, 957-974.
P Agarwal, SSever Dragomir, M Jleli and B Samet. Advances in mathematical inequalities and applications. Trends in Mathematics. Birkhauser, Switzerland, 2018.
Q Sheng, D Liu and J Zheng. NiCo alloy nanoparticles anchored on polypyrrole/reduced graphene oxide nanocomposites for nonenzymatic glucose sensing. New Journal of Chemistry 2016; 40(8), 6658-6665.
U Paramo-Garcia, A Avalos-Perez, J Guzman-Pantoja, NP Diaz-Zavala, JA MeloBanda, NV Gallardo-Rivas, J Reyes-Gomez, D Pozas-Zepeda, JG Ibanez and N Batina. Polypyrrole microcontainer structures and doughnuts designed by electrochemical oxidation: An electrochemical and scanning electron microscopy study. E-Polymers 2014; 14(1), 75-84.
N Zaini, NSM Hanapi, WNW Ibrahim, R Osman, S Kamaruzaman, N Yahaya and AL Anis. Dispersive micro-solid-phase extraction (D-µ-SPE) with polypyrrole-graphene oxide (PPy-GO) nanocomposite sorbent for the determination of tetracycline antibiotics in water samples. Malaysian Journal of Analytical Sciences 2022; 26(5), 953-964.
S Konwer, R Boruah and SK Dolui. Studies on conducting polypyrrole/graphene oxide composites as supercapacitor electrode. Journal of Electronic Materials 2011; 40, 2248-2255.
J Wu, W Pisula and K Mullen. Graphenes as potential material for electronics. Chemical Reviews 2007; 107, 718-747.
NF Attia, SM Lee, HJ Kim and KE Geckeler. Nanoporous polypyrrole: Preparation and hydrogen storage properties. International Journal of Energy Research 2014; 38(4), 466-476.
T Esfandiyari, N Nasirizadeh, M Dehghania and MH Ehrampoosh. Graphene oxide-based carbon composite as adsorbent for Hg removal: Preparation, characterization, kinetics and isotherm studies. Chinese Journal of Chemical Engineering 2017; 25(9), 1170-1175.
Z Liu, Z Gao, L Xu and F Hu. Polypyrrole modified magnetic reduced graphene oxide composites: Synthesis, characterization and application for selective lead adsorption. RSC Advances 2020; 10(30), 17524-17533.
S Anuma, P Mishra and BR Bhat. Polypyrrole functionalized cobalt oxide graphene (COPYGO) nanocomposite for the efficient removal of dyes and heavy metal pollutants from aqueous effluents. Journal of Hazardous Materials 2021; 416, 125929.
J Stejskal, M Kohl, M Trchova, Z Kolska, M Pekarek, I Krivka and J Prokes. Conversion of conducting polypyrrole nanostructures to nitrogen-containing carbons and its impact on the adsorption of organic dye. Material Advances 2021; 2(2), 706-717.
A Moyseowicz and G Gryglewicz. High-performance hybrid capacitor based on a porous polypyrrole/reduced graphene oxide composite and a redox-active electrolyte. Electrochimica Acta 2020; 354, 136661.
P Pattananuwat, R Pornprasertsuk, J Qin and S Prasertkaew. Polypyrrole nanoparticles embedded nitrogen-doped graphene composites as novel cathode for long life cycles and high-power zinc-ion hybrid supercapacitors. RSC Advances 2021; 11(56), 35205-35214.
S Kalam, SA Abu-Khamsin, MS Kamal and S Patil. Surfactant adsorption isotherms: A review. ACS Omega 2021; 6(48), 32342-32348.
T Li, Z Jiang, C Xu, B Liu, G Liu, P Wang, X Li, W Chen, C Ning and Z Wang. Effect of pore structure on shale oil accumulation in the lower third member of the Shahejie formation, Zhanhua Sag, eastern China: Evidence from gas adsorption and nuclear magnetic resonance. Marine and Petroleum Geology 2017; 88, 932-949.
MA Gado. Sorption of thorium using magnetic graphene oxide polypyrrole composite synthesized from natural source. Separation Science and Technology 2018; 53(13), 2016-2033.
T Sudha, G Divya, J Sujaritha and P Duraimurugan. Review of experimental design in analytical chemistry. Indo American Journal of Pharmaceutical Research 2017; 7(8), 550-565.
M Chen, Z Yan, J Luan, X Sun, W Liu and X Ke. π-π electron-donor-acceptor (EDA) interaction enhancing adsorption of tetracycline on 3D PPY/CMC aerogels. Chemical Engineering Journal 2023; 454(4), 140300.
H Sereshti, S Soltani, N Sridewi, E Salehi, E Parandi, HR Nodeh and S Shahabuddin. Solid phase extraction penicillin and tetracycline in human serum using magnetic graphene oxide-based sulfide nanocomposite. Magnetochemistry 2023; 9(5), 132.
PN Sharma, G Shmueli, M Sarstedt, N Danks and S Ray. Prediction‐oriented model selection in partial least squares path modeling. Decision Sciences 2021; 52(3), 567-607.
D Chicco, MJ Warrens and G Jurman. The coefficient of determination R-squared is more informative than SMAPE, MAE, MAPE, MSE and RMSE in regression analysis evaluation. Peerj Computer Science 2021; 7(3), 623.
M Ruzhansky, YJ Cho, P Agarwal and I Area. Advances in real and complex analysis with applications. Birkhauser, Switzerland, 2017.
K Murtada and J Pawliszyn. Evolution in understandings of the design, optimization, and application of sorbent-based extraction devices. TrAC Trends in Analytical Chemistry 2023; 169, 117396.
JC Cruz, IDD Souza, CF Grecco, EC Figueiredo and MEC Queiroz. Recent advances in column switching high-performance liquid chromatography for bioanalysis. Sustainable Chemistry and Pharmacy 2021; 21, 100431.
OB Rudakov, LV Rudakova and VF Selemenev. Acetonitrile as tops solvent for liquid chromatography and extraction. Journal of Analytical Chromatography and Spectroscopy 2023; 6, 883.
S Fekete, JL Veuthey and D Guillarme. Comparison of the most recent chromatographic approaches applied for fast and high-resolution separations: Theory and practice. Journal of Chromatography A 2015; 1408, 1-14.
SK Muchakayala, NK Katari, KK Saripella, H Schaaf, VM Marisetti, SK Ettaboina and VK Rekulapally. Implementation of analytical quality by design and green chemistry principles to develop an ultra-high performance liquid chromatography method for the determination of Fluocinolone Acetonide impurities from its drug substance and topical oil formulations. Journal of Chromatography A 2022; 1697, 463380.
SW Foster, N Wright, JP Grinias and LM Blumberg. Measurement of optimal flow rate in gradient elution liquid chromatography. Journal of Chromatography A 2021; 1659, 462645.
Z Li, H Jiang, X Wang, C Wang and X Wei. Effect of pH on adsorption of tetracycline antibiotics on graphene oxide. International Journal of Environmental Research and Public Health 2023; 20(3), 2448.
S Casado‐Terrones, A Segura‐Carretero, S Busi, G Dinelli and A Fernandez‐Gutierrez. Determination of tetracycline residues in honey by CZE with ultraviolet absorbance detection. Electrophoresis 2007; 28(16), 2882-2887.
R LoBrutto, A Jones, YV Kazakevich and HM McNair. Effect of the eluent pH and acidic modifiers in high-performance liquid chromatography retention of basic analytes. Journal of Chromatography A 2001; 913(1-2), 173-187.
JJ Kirkland and JJ DeStefano. The art and science of forming packed analytical high-performance liquid chromatography columns. Journal of Chromatography A 2006; 1126(1-2), 50-57.
JP Maran, S Manikandan, B Priya and P Gurumoorthi. Box-Behnken design based multi-response analysis and optimization of supercritical carbon dioxide extraction of bioactive flavonoid compounds from tea (Camellia sinensis L.) leaves. Journal of Food Science and Technology 2015; 52, 92-104.
S Ncube, LM Madikizela, MM Nindi and L Chimuka. Solid phase extraction technique as a general field of application of molecularly imprinted polymer materials. Comprehensive Analytical Chemistry 2019; 86, 41-76.
F Wu, A Xie, M Sun, Y Wang and M Wang. Reduced graphene oxide (RGO) modified spongelike polypyrrole (PPy) aerogel for excellent electromagnetic absorption. Journal of Materials Chemistry A 2015; 3(27), 14358-14369.
I Vergara-Luis, JC Baez-Millan, I Baciero, B Gonzalez-Gaya, M Olivares, O Zuloaga and A Prieto. Comparison of conventional and dispersive solid phase extraction clean-up approaches for the simultaneous analysis of tetracyclines and sulfonamides in a variety of fresh vegetables. Talanta 2023; 254, 124192.
K Meftah, S Meftah, H Lamkhanter, T Bouzid, Y Rezzak, S Touil and A Abid. Extraction and optimization of Austrocylindropuntia subulata powder as a novel green coagulant. Desalination and Water Treatment 2024; 318, 100339.
S Meftah, K Meftah, M Drissi, I Radah, K Malous, A Amahrous, A Chahid, T Tamri, A Rayyad, B Darkaoui, S Hanine, O El‑Hassan and L Bouyazza. Heavy metal polluted water: Effects and sustainable treatment solutions using bio‑adsorbents aligned with the SDGs. Discover Sustainability 2025; 6, 137.
K Meftah, S Meftah, H Lamkhanter, I Hamzaoui, S Touil and A Abid. Green flocculant performance for wastewater treatment using cactus plant powder: A mini‑review. Journal of Analytical Sciences and Applied Biotechnology 2023; 5(1), 29-34.
S Meftah, K Meftah, H Lamkhanter, T Mehdi, A Amahrouss, F Assadid, O El Hassan and L Bouyazza. The main types of derivatives of plant matter and agricultural waste used as bio‑adsorbents for the removal of heavy metals and dyes: A review. Egyptian Journal of Chemistry 2024; 67(9), 185-207.
C Tu, Y Dai, K Xu, M Qi, W Wang, L Wu and A Wang. Determination of tetracycline in water and honey by iron (II, III)/aptamer-based magnetic solid-phase extraction with high-performance liquid chromatography analysis. Analytical Letters 2019; 52(1), 1653-1669.
H Sereshti, F Karami and N Nouri. A green dispersive liquid-liquid microextraction based on deep eutectic solvents doped with β-cyclodextrin: Application for determination of tetracyclines in water samples. Microchemical Journal 2021; 163, 105914.
L Lian, J Lv, X Wang and D Lou. Magnetic solid–phase extraction of tetracyclines using ferrous oxide coated magnetic silica microspheres from water samples. Journal of Chromatography A 2018; 1543, 1-9.
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