Prepatrion of the patentiometric immobilizied urease electrode anf urea determination
Küçük Resim Yok
Tarih
1996
Yazarlar
Dergi Başlığı
Dergi ISSN
Cilt Başlığı
Yayıncı
Ege Üniversitesi
Erişim Hakkı
info:eu-repo/semantics/closedAccess
Özet
11 ÖZET POTANSÎYOMETRÎK ESASLI IMMOBÎLÎZE UREAZ ELEKTRODU HAZIRLANMASI VE ÜRE TAYÎNÎ ŞEHlTO?ULLARI, Ahmet Yüksek Lisans Tezi, Biyokimya Bölümü Tez Yöneticisi : Prof.Dr. Arif H.USLAN Kasım 1996, 47 sayfa Bu tezde, PVA(polyvinilalkol), FMP(2-Floro-l-metilpiridinium tolien-4-sülfonat) ile aktifleştirilmiş ve üreaz (EC 3.5.1.5) kovalent olarak bağlanmıştır. PVA-Üreaz gluteraldehit yardımıyla jelatin ile çapraz bağlanarak bir pH cam elektrodu üzerinde immobilize edilerek üreye duyarlı bir enzim elektrodu geliştirilmiştir. Ölçümler enzimatik reaksiyon uyarınca, üre konsantrasyonuyla orantılı olarak değişen ortam potansiyelinin belirlenmesi yoluyla ve çizilen standart grafikler yardımıyla yapılmıştır. Hazırlanan biyosensörün optimiasyon çalışmalarında en uygun üreaz, jelatin ve gluteraldehit miktarları sırasıyla, 8 mg/ml, 54 mg/ml ve 0.5% ile 5.0 dak. olarak bulundu. Optimum çalışma ortamı olarak 10 mM'lık pH:8.0 olan fosfat tamponunun kullanılması ve ölçümlerin 25°C yapılması gerektiği sonucuna varıldı. Geliştirilen biyosensörün karakterizasyonu amacıyla yapılan çalışmalar sonucunda 8.9xl0~5-l"lxl0~3 M üre konsantrasyon aralığında doğrusal sonuçların alınabildiği tesbit edildi. Ayrıca tampon konsantrasyonunu, 10 mM-100 mM arasında değiştirerek 10-2-10-5 yi jjre konsantrasyonu aralığında çalışılabileceği tesbit edildi. Tekrarlanabilirlik denemesi (n=20), aynı bir serum örneği için üreaz enzim elektrodu ve fotometrik metod ile yapılmıştır. Her iki metod için sırasıyla ortalama değerler (X)ı=5.96 mM ve (X)2=5.86 mM, standart sapmalar (S.S)ı=±0.149 ve (S.S)2=±0.206, varyasyon katsayıları (V.K)ı=%2.5 ve (V.K)2=%3.51 olarak bulunmuştur. Bu sonuçlara göre üreaz enzim elektrodunun daha hassas ve duyarlı olduğu görülmektedir. Elektrod için depo kararlılığı araştırılmıştır. Anahtar Kelimeler: Üreaz, Üre Tayini, Enzim Elektrodu, Biyosensörler
İÜ ABSTRACT PREPARATION OF THE POTENTIOMETRIC IMMOBILIZED UREASE ELECTRODE AND UREA DETERMINATION ŞEHÎTO?ULLARI, Ahmet MSc in Biochemistry Supervisor : Prof.Dr. Arif H.USLAN November 1996, 47 page In this thesis, PVA (polyvinylalcohol) was activated with FMP (2-Fluoro-l-methylpyridiumtoluene-4-sulphonate) and urease (EC.3.5.1.5) was linked to the activated substance covalently. PVA-Urease was cross-linked with gelatine by using gluteraldehyde and was immobilized on a surface of pH-glass electrode, therefore, enzyme electrode which was sensitive for urea was developed. Measurements were carriede out by standard curves which were determined by the changing medium potential related to urea concentration in the enzymatic reaction. In the optimization studies of biosensors prepared, the most suitable urease, gelatine and gluteraldehyde amounts were determined as 8 mg/ml, 54 mg/ml and for 5 min. 0.5 % respectively. The phosphate buffer (pH:8.0, 10 mM) and 25°C were established as providing the optimum conditions. The characterization of the developed biosensor studies proved a linearity in the urea concentration range 8.9x10"^- 1.1x10" 3 M. Also, by changing the buffer concentration (10 mM-100 mM range), it was stablied that can be studied at the urea concentration range of 10"^-10"5 M. Reproducibilty experiments (n=20) were done with urease enzyme electrode and photometric method for the same serum sample. Average valves [(X)ı,(X)2İ for each two methods were 5.96 mM and 5.86 mM, standard deviations [(S.D)j,(S.D)2] were ± 0.149 and ±0.206 and variation coefficients [(C.V)i,(C.V)2] were 2.50% and 3.51%, respectively. According to these results, it was observed that urease electrode was more sensitive. Storage stability were investigated for the urease enzyme electrode. Key Words: Urease, Urea Determination, Enzyme Electrodes, Biosensors
İÜ ABSTRACT PREPARATION OF THE POTENTIOMETRIC IMMOBILIZED UREASE ELECTRODE AND UREA DETERMINATION ŞEHÎTO?ULLARI, Ahmet MSc in Biochemistry Supervisor : Prof.Dr. Arif H.USLAN November 1996, 47 page In this thesis, PVA (polyvinylalcohol) was activated with FMP (2-Fluoro-l-methylpyridiumtoluene-4-sulphonate) and urease (EC.3.5.1.5) was linked to the activated substance covalently. PVA-Urease was cross-linked with gelatine by using gluteraldehyde and was immobilized on a surface of pH-glass electrode, therefore, enzyme electrode which was sensitive for urea was developed. Measurements were carriede out by standard curves which were determined by the changing medium potential related to urea concentration in the enzymatic reaction. In the optimization studies of biosensors prepared, the most suitable urease, gelatine and gluteraldehyde amounts were determined as 8 mg/ml, 54 mg/ml and for 5 min. 0.5 % respectively. The phosphate buffer (pH:8.0, 10 mM) and 25°C were established as providing the optimum conditions. The characterization of the developed biosensor studies proved a linearity in the urea concentration range 8.9x10"^- 1.1x10" 3 M. Also, by changing the buffer concentration (10 mM-100 mM range), it was stablied that can be studied at the urea concentration range of 10"^-10"5 M. Reproducibilty experiments (n=20) were done with urease enzyme electrode and photometric method for the same serum sample. Average valves [(X)ı,(X)2İ for each two methods were 5.96 mM and 5.86 mM, standard deviations [(S.D)j,(S.D)2] were ± 0.149 and ±0.206 and variation coefficients [(C.V)i,(C.V)2] were 2.50% and 3.51%, respectively. According to these results, it was observed that urease electrode was more sensitive. Storage stability were investigated for the urease enzyme electrode. Key Words: Urease, Urea Determination, Enzyme Electrodes, Biosensors
Açıklama
Bu tezin, veri tabanı üzerinden yayınlanma izni bulunmamaktadır. Yayınlanma izni olmayan tezlerin basılı kopyalarına Üniversite kütüphaneniz aracılığıyla (TÜBESS üzerinden) erişebilirsiniz.
Anahtar Kelimeler
Biyomühendislik, Bioengineering, Üre, Urea, Üreaz, Urease
