Sefazidim Analizi İçin GO@AuNPs Destekli Moleküler Baskılı Polimer Temelli Elektrokimyasal Sensör Tasarımı

Abstract

Üçüncü nesil sefalosporin olan seftazidim (CTZ), Gram-negatif patojenlere karşı güçlü aktivitesi nedeniyle ciddi bakteriyel enfeksiyonların tedavisinde yaygın olarak kullanılmaktadır. Biyolojik ve farmasötik matrislerde CTZ'nin doğru ve hassas bir şekilde belirlenmesi, terapötik ilaç izleme ve kalite kontrolü için büyük önem taşımaktadır. Bu çalışmada, CTZ'nin seçici ve ultra hassas tespiti için moleküler baskılı polimer (MIP) temelli bir elektrokimyasal sensör tasarlanmıştır. Mükemmel elektriksel iletkenliği, geniş özgül yüzey alanı ve sinerjik sinyal yükseltmesi nedeniyle destekleyici matris olarak grafen oksit/altın nanopartikül (GO@AuNPs) nanokompoziti kullanılmıştır. Bu nanoyapılı platformun entegrasyonu, sensörün elektrokimyasal performansını önemli ölçüde artırmıştır. Moleküler tanıma, GO@AuNPs ile modifiye edilmiş elektrot yüzeyine bir MIP tabakası oluşturularak sağlanmıştır. Polimerik film, polimer yapısının hassas kontrolüne olanak tanıyan ve hafif reaksiyon koşulları altında yapısal homojenliği sağlayan fotopolimerizasyon (PP) yoluyla sentezlenmiştir. Trans-3-(3-piridil)akrilik asit (3,3-PAA), CTZ ile güçlü ve spesifik etkileşimleri nedeniyle seçilen fonksiyonel monomer olarak kullanıldı ve böylece baskılı polimer ağında yüksek afinite ve seçiciliğe sahip tamamlayıcı tanıma bölgelerinin oluşumunu kolaylaştırdı. Optimize edilmiş deneysel koşullar altında, geliştirilen MIP sensörü, düşük tespit limiti (LOD) ile geniş bir doğrusal yanıt aralığı sergileyerek CTZ'ye karşı mükemmel hassasiyet gösterdi. Seçicilik çalışmaları, sensörün yapısal olarak benzer sefalosporin antibiyotiklerine karşı yüksek özgüllüğünü doğruladı. Ayrıca, önerilen sensörün pratik uygulanabilirliği, gerçek biyolojik örneklerde ve farmasötik formülasyonlarda CTZ'nin başarılı bir şekilde tespit edilmesiyle doğrulandı ve gerçek örnek analizi için sağlamlığı ve potansiyeli vurgulandı.

Keywords

• Sefazidim
• Moleküler baskılı polimerler
• İlaç analizi
• Nanomalzemeler
• Elektrokimyasal sensörler

Design of a GO@AuNPs-embedded MIP-based electrochemical sensor for ceftazidime analysis

Abstract

Ceftazidime (CTZ), a third-generation cephalosporin, is widely used to treat severe bacterial infections owing to its strong activity against Gram-negative pathogens. Accurate and sensitive determination of CTZ in biological and pharmaceutical matrices is of great importance for therapeutic drug monitoring and quality control. In this study, a molecularly imprinted polymer (MIP)–based electrochemical sensor was designed for the selective and ultrasensitive detection of CTZ. A graphene oxide/gold nanoparticle (GO@AuNPs) nanocomposite was used as the supporting matrix owing to its excellent electrical conductivity, large specific surface area, and synergistic signal amplification. The incorporation of this nanostructured platform significantly enhanced the sensor's electrochemical performance. Molecular recognition was achieved by constructing an MIP layer on the GO@AuNPs-modified electrode surface. The polymeric film was synthesized via photopolymerization (PP), which allows precise control of polymer structure and ensures structural uniformity under mild reaction conditions. Trans-3-(3-pyridyl)acrylic acid (3,3-PAA) was utilized as the functional monomer, selected for its strong and specific interactions with CTZ, thereby facilitating the formation of complementary recognition sites with high affinity and selectivity within the imprinted polymer network. Under optimized experimental conditions, the sensor exhibited a linear response in the range of 2.50×10⁻¹³ to 3.75×10⁻¹² M, with a limit of detection (LOD) of 5.65×10⁻¹⁴ M and a limit of quantification (LOQ) of 1.88×10⁻¹³ M. Selectivity studies demonstrated negligible interference even in the presence of a 1000-fold excess of structurally related compounds. The method was successfully applied to commercial serum samples and pharmaceutical dosage forms, yielding recovery values of 98.61–99.82% and 99.60%, respectively. Furthermore, the practical applicability of the proposed sensor was validated by successfully CTZ in real biological samples and pharmaceutical formulations, highlighting its robustness and potential for real-sample analysis.

Keywords

• Ceftazidime
• Molecularly imprinted polymers
• Drug analysis
• Nanomaterials
• Electrochemical sensors

References

1. Rains, C. P., Bryson, H. M., & Peters, D. H. (1995). Ceftazidime: An update of its antibacterial activity, pharmacokinetic properties and therapeutic efficacy. Journal of Pharmaceutical and Biomedical Analysis, 49(4), 577–617.
2. Kontou, A., Kourti, M., Iosifidis, E., Sarafidis, K., & Roilides, E. (2023). Use of newer and repurposed antibiotics against gram-negative bacteria in neonates. Antibiotics, 12(6), 1072.
3. Myers, C. M., & Blumer, J. L. (1983). Determination of ceftazidime in biological fluids by using high-pressure liquid chromatography. Antimicrobial Agents and Chemotherapy, 24(3), 343–346.
4. Legrand, T., Vodovar, D., Tournier, N., Khoudour, N., & Hulin, A. (2016). Simultaneous determination of eight β-lactam antibiotics in human plasma by ultra-high-performance liquid chromatography with ultraviolet detection. Antimicrobial Agents and Chemotherapy, 60(8), 4734–4742.
5. Cazorla-Reyes, R., Romero-González, R., Frenich, A. G., Maresca, M. A. R., & Vidal, J. L. M. (2014). Simultaneous analysis of antibiotics in biological samples by ultra-high-performance liquid chromatography–tandem mass spectrometry. Journal of Pharmaceutical and Biomedical Analysis, 89, 203–212.
6. Lefeuvre, S., Bois-Maublanc, J., Hocqueloux, L., Bret, L., Francia, T., et al. (2017). A simple ultra-high-performance liquid chromatography–high resolution mass spectrometry assay for the simultaneous quantification of 15 antibiotics in plasma. Journal of Chromatography B, 1065, 50–58.
7. de Haro Moreno, A., & Salgado, H. R. N. (2012). Development and validation of the quantitative analysis of ceftazidime in powder for injection by infrared spectroscopy. Physical Chemistry, 2, 6–11.
8. de Haro Moreno, A., & Salgado, H. R. N. (2009). Rapid and selective UV spectrophotometric method for the analysis of ceftazidime. Journal of AOAC International, 92(3), 820–823.

9. Bergman, J., Harvill, L., Hawkins, S., Sladky, K., & Cox, S. (2021). Determination of ceftazidime in plasma by RP-HPLC and ultraviolet detection. Biomedical Chromatography, 35(7), e5104.
10. Xie, T., Wang, L., Wang, H., Cao, C., Tang, C., et al. (2024). In-situ electrodeposition of Co/Ce-MOF-derived carbon composites as an anode for electrochemical degradation of ceftazidime. Separation and Purification Technology, 330, 125231.
11. Ni, Y., Kang, Y., Liu, Y., Bi, W., Qin, J., et al. (2024). Fabrication and characterization of a novel Ce-modified PbO₂ electrode with Ti₄O₇-rGO as a middle layer for the degradation of ceftazidime. Desalination, 117645.
12. Li, X., Duan, P., Lei, J., Sun, Z., & Hu, X. (2019). Fabrication of Ti/TiO₂/SnO₂-Sb-Cu electrode for enhancing electrochemical degradation of ceftazidime in aqueous solution. Journal of Electroanalytical Chemistry, 847, 113231.
13. Scholz, F., Stojek, Z., Inzelt, G., et al. (2010). Electroanalytical Methods: Guide to experiments and applications (2nd ed.). Springer, Berlin, Heidelberg, XXVII, 359.
14. Baig, N., Sajid, M., & Saleh, T. A. (2019). Recent trends in nanomaterial-modified electrodes for electroanalytical applications. TrAC - Trends in Analytical Chemistry, 111, 47–61.
15. Zabihollahpoor, A., Rahimnejad, M., Najafpour-Darzi, G., & Moghadamnia, A. A. (2020). Recent advances in electroanalytical methods for the therapeutic monitoring of antiepileptic drugs: A comprehensive review. Journal of Pharmaceutical and Biomedical Analysis, 188, 113394.
16. Chaudhary, M., Kumar, A., Devi, A., Singh, B. P., Malhotra, B. D., & Singhal, K. (2023). Prospects of nanostructure-based electrochemical sensors for drug detection: A review. Materials Advances, 4, 432–457.
17. De Rycke, E., Stove, C., Dubruel, P., De Saeger, S., & Beloglazova, N. (2020). Recent developments in electrochemical detection of illicit drugs in diverse matrices. Biosensors and Bioelectronics, 169, 112579.
18. Klimuntowski, M., Alam, M. M., Singh, G., & Howlader, M. M. R. (2020). Electrochemical sensing of cannabinoids in biofluids: A noninvasive tool for drug detection. ACS Sensors, 5(3), 620–636.
19. Cetinkaya, A., Kaya, S. I., Corman, M. E., Karakaya, M., Atici, E. B., & Ozkan, S. A. (2022). A highly sensitive and selective electrochemical sensor based on computer-aided design of molecularly imprinted polymer for the determination of leflunomide. Microchemical Journal, 179, 107496.
20. Piskin, E., Cetinkaya, A., Eryaman, Z., Karadurmus, L., Unal, M. A., et al. (2024). Development of ultra-sensitive and selective molecularly imprinted polymer-based electrochemical sensor for L-lactate detection. Microchemical Journal, 204, 111163.
21. Yence, M., Cetinkaya, A., Çorman, M. E., Uzun, L., Caglayan, M. G., et al. (2023). Fabrication of molecularly imprinted electrochemical sensors for sensitive codeine detection. Microchemical Journal, 193, 109060.
22. Çorman, M. E., Cetinkaya, A., Armutcu, C., Uzun, L., & Ozkan, S. A. (2023). Designing of ZnO nanoparticles-oriented interface imprinted electrochemical sensor for fluoxetine detection. Bioelectrochemistry, 152, 108411.
23. Cetinkaya, A., Unal, M. A., Nazir, H., Corman, M. E., Uzun, L., et al. (2024). Development of borazine-assisted-oriented molecularly imprinted electrochemical sensor for the detection of umifenovir in serum and urine. Sensors and Actuators B: Chemical, 420, 136519.
24. Cetinkaya, A., Unal, M. A., Nazır, H., Çorman, M. E., Uzun, L., & Ozkan, S. A. (2024). A comparative study of electropolymerization and photopolymerization for the determination of molnupiravir via molecularly imprinted polymers. Microchimica Acta, 191(5), 270.
25. Li, X., Wang, X., Li, L., Duan, H., & Luo, C. (2015). Electrochemical sensor based on magnetic graphene oxide@gold nanoparticles–molecular imprinted polymers for determination of dibutyl phthalate. Talanta, 131, 354–360.
26. Huang, Z., Liu, T., Geng, J., Zhang, S., Neha, Ye, M., & Sun, W. (2026). Molecularly imprinted electrochemical sensor based on AuNPs-nitrogen doped graphene-MXene nanocomposite for the detection of kanamycin in milk. Journal of Food Composition and Analysis, 108868.
27. Deepa, J. R., Jyothish, B., Athira, V. S., Mini, S., Sree Remya, T. S., & Nair, A. A. (2025). Molecularly imprinted electrochemical sensor enhanced with functionalized gold nanoparticle/graphene oxide composite for ultra-selective detection of homocysteine. Microchemical Journal, 115310.
28. Mani, A., & Anirudhan, T. S. (2024). Electrochemical sensing of cortisol by gold nanoparticle incorporated carboxylated graphene oxide based molecularly imprinted polymer. Chemical Engineering Journal, 152654.
29. Zhong, Y., Li, Z., Zhang, A., Peng, Y., Zhou, H., et al. (2024). Gold nanoparticle mediated molecularly imprinted electrochemical sensor for detection of neutral phosmet residues. Microchemical Journal, 201, 110728.
30. Mani, A., & Anirudhan, T. S. (2024). Electrochemical sensing of cortisol by gold nanoparticle incorporated carboxylated graphene oxide based molecularly imprinted polymer. Chemical Engineering Journal, 493, 152654.
31. Chauhan, V., Sharma, M., Tiwari, A., & et al. (2024). Developing, validating, and comparing an analytical method to simultaneously detect z‑drugs in urine samples using the QuEChERS approach with both liquid chromatography and gas chromatography‑tandem mass spectrometry. Saudi Pharmaceutical Journal, 32(2), 101950.
32. Honert, C., Wifling, K., Lazo Hernández, M. J., & al. (2025). Assessment of current use pesticides in flowers, pollen provision, and wild bees: HPLC‑ESI‑MS/MS method development and field implementation. Analytical and Bioanalytical Chemistry, 417, 4199‑4213.
33. Koziarska, M., Strzebońska, M., & Szalińska, E. (2025). Development and validation of a green/blue UHPLC‑MS/MS method for trace pharmaceutical monitoring. Scientific Reports, 15, 15614.
34. Askar, A. M., Al Ali, A. Y., Khalifa, M. K., Salem, A. A., Alkhuwaildi, B. M., & Shah, I. (2025). Rapid GC‑MS method for screening seized drugs in forensic investigations: optimization and validation. Frontiers in Chemistry, 13, 1559279.
35. A. Procedures, Guidance for Industry Q2B Validation of Analytical Procedures: Methodology, 20857, 301–827.
36. Ozkan, S. A., Kauffmann, J.-M., & Zuman, P. (2015). Electroanalytical method validation in pharmaceutical analysis and Their Applications. In S.A. Ozkan, J.M. Kauffmann, P. Zuman (Eds) Electroanalysis in biomedical and pharmaceutical sciences, (1st ed., pp. 235-266). Springer, Berlin, Heidelberg.