Ürik asit tayinine yönelik moleküler baskılanmış yüzey plazmon rezonans sensör hazırlanması
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Date
2016-03-11
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Uludağ Üniversitesi
Abstract
Bu çalışmada, ürik asit (UA) tayini için tanıma aracı olarak moleküler baskılanmış nanopartiküllerin kullanıldığı yüzey plazmon rezonans (SPR) sensör hazırlanmıştır. Sensör, SPR sensörün altın yüzeyinin UA baskılanmış poli(hidroksietil metakrilatmetakriloil- amido-sistein metil ester)-Fe3+ [poli(HEMA-MAC)-Fe3+] nanopartiküller ile modifiye edilmesi ile hazırlandı. Fonksiyonel monomer (MAC) ve kalıp molekül (UA) arasındaki ön-organizasyon metal-şelat etkileşimi ile gerçekleştirildi. Metal iyonu olarak Fe3+ iyonu kullanıldı. Hazırlanan MAC-Fe3+-UA kompleksi FTIR tekniği kullanılarak karakterize edildi. Poli(HEMA-MAC)-Fe3+ nanopartiküller FTIR, elementel analiz, FE-SEM, CTEM, EDS, zeta boyut dağılımı analizleri ile karakterize edildi. UA baskılanmış poli(HEMA-MAC)-Fe3+ SPR sensör nanopartikül çözeltisinin altın yüzeye damlatılması ve ardından kurutulması ile hazırlandı. Hazırlanan SPR sensör AFM, temas açısı ve optik profilometre analizleri ile karakterize edildi. SPR sensörün ürik asit tayin kapasitesi farklı derişimlerdeki ürik asit çözeltilerinin (0,5-40 mg/L) SPR sensör ile etkileştirilmesi ile belirlendi. Elde edilen sensör cevapları ile bağlanma, kinetik ve izoterm analizleri yapıldı. Ayrıca hazırlanan sensörün LOD (tespit limiti) 0,247 mg/L ve LOQ (tayin limiti) 0,825 mg/L olarak hesaplandı. SPR sensörün seçiciliği askorbik asit, teofilin ve üre kullanılarak araştırıldı. Aynı zamanda ikili, üçlü ve dörtlü karışımlar hazırlanarak yarışmalı kinetik analizler yapıldı. UA baskılanmış SPR sensör idrarda UA tayini için kullanıldı ve LOD ve LOQ değerleri sırasıyla 0,498 mg/L ve 1,66 mg/L olarak belirlendi. Analiz sonuçları standart UA tayin yöntemiyle karşılaştırıldı. Hazırlanan sensörün idrardaki UA derişimini % 97,73 doğrulukta tayin edebildiği belirlendi. Poli(HHEMA-MAC)-Fe3+ SPR sensörün ürik asit tayininde tekrar kullanılabilirliği araştırıldı.
In this study, SPR sensor was prepared that use molecular imprinted nanoparticles as molecular recognition element for uric acid dedection. SPR sensor was prepared by modification of the gold surface of SPR sensor with UA imprinted poly(hydroxyethyl methacrylate-methacryloyl-amido-cysteine methyl ester)-Fe3+ [poly(HEMA-MAC)- Fe3+] nanoparticles. Preorganization of functional monomer (MAC) and template (UA) was provided by using metal-chelate interaction. Fe3+ was the metal ion. MAC-Fe3+-UA complex was characterized with FTIR analysis. The characterization of poly(HEMAMAC)- Fe3+ nanoparticles were done by FTIR, elemental analysis, SEM, TEM, EDS, zeta size analysis. The UA imprinted SPR sensor was obtained by dropping and drying a small amount of nanoparticle solution to gold surface. Prepared SPR sensor was characterized with AFM, contact angle and optic profilometer measurements. UA sensing ability of imprinted SPR sensor was investigated by interacting UA solutions in different concentrations (0.5-40 mg/L) with SPR sensor. The limit of detection (LOD) and limit of quantation (LOQ) were calculated for prepared SPR sensor as 0.247 mg/L and 0.825 mg/L, respectively. In order to show the selectivity of the UA imprinted poly(HEMA-MAC)-Fe3+ SPR sensor, ascorbic acid, theophylline and urea were used. In addition, binary, triple and quartet mixtures of the molecules were prepared for competitive kinetic analysis. UA-imprinted SPR sensor was used for UA determination in urine. LOD and LOQ values were calculated as 0.498 mg/L and 1.66 mg/L, respectively. The results were compared with standart method. The prepared sensor measured the UA concentration 97.73 % accuracy. The reusability of the poli(HEMAMAC)- Fe3+ SPR sensor was also investigated.
In this study, SPR sensor was prepared that use molecular imprinted nanoparticles as molecular recognition element for uric acid dedection. SPR sensor was prepared by modification of the gold surface of SPR sensor with UA imprinted poly(hydroxyethyl methacrylate-methacryloyl-amido-cysteine methyl ester)-Fe3+ [poly(HEMA-MAC)- Fe3+] nanoparticles. Preorganization of functional monomer (MAC) and template (UA) was provided by using metal-chelate interaction. Fe3+ was the metal ion. MAC-Fe3+-UA complex was characterized with FTIR analysis. The characterization of poly(HEMAMAC)- Fe3+ nanoparticles were done by FTIR, elemental analysis, SEM, TEM, EDS, zeta size analysis. The UA imprinted SPR sensor was obtained by dropping and drying a small amount of nanoparticle solution to gold surface. Prepared SPR sensor was characterized with AFM, contact angle and optic profilometer measurements. UA sensing ability of imprinted SPR sensor was investigated by interacting UA solutions in different concentrations (0.5-40 mg/L) with SPR sensor. The limit of detection (LOD) and limit of quantation (LOQ) were calculated for prepared SPR sensor as 0.247 mg/L and 0.825 mg/L, respectively. In order to show the selectivity of the UA imprinted poly(HEMA-MAC)-Fe3+ SPR sensor, ascorbic acid, theophylline and urea were used. In addition, binary, triple and quartet mixtures of the molecules were prepared for competitive kinetic analysis. UA-imprinted SPR sensor was used for UA determination in urine. LOD and LOQ values were calculated as 0.498 mg/L and 1.66 mg/L, respectively. The results were compared with standart method. The prepared sensor measured the UA concentration 97.73 % accuracy. The reusability of the poli(HEMAMAC)- Fe3+ SPR sensor was also investigated.
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Keywords
Moleküler baskılama, Yüzey plazmon rezonans, Ürik asit, Biyosensör, Molecular imprinting, Surface plasmon resonance, Uric acid, Biosensor
Citation
Sarıkaya, A. G. (2016). Ürik asit tayinine yönelik moleküler baskılanmış yüzey plazmon rezonans sensör hazırlanması. Yayınlanmamış doktora tezi. Uludağ Üniversitesi Fen Bilimleri Enstitüsü.