Thesis Type: Post Graduate
Institution Of The Thesis: Gazi Üniversitesi, Sağlık Bilimleri Enstitüsü, Turkey
Approval Date: 2010
Student: DENİZ KOÇDAN
Consultant: HASAN BASANAbstract:
In this study, a spectroflourimetric method was developed for the determination of calcitonin in ampules. For this purpose, calcitonin was derivatized using fluorescamine as a fluorogenic agent. After optimization of the derivatization conditions, it was decided to use 5.0 mL of 0.5 M borate buffer pH 8.5. Fluorescamine was dissolved in acetone, 0.125 mg/mL. In order to see whether derivatization reaction was achieved or not, absorption spectrum of derivatized sCT was recorded using UV/VIS spectrophotometer and it was seen that there was an absorption maximum at 389 nm conforming the derivatization. Quantitative determinations were conducted by setting the excitation and emission monochromators and slit widths to 389, 484 and 10 nm, respectively. Standard solutions of the derivatized sCT were in the range of 0.5 to 6.0 μg/mL and regression equation and correlation coefficient were obtained as F=120 C +24.7 and R = 0.9990, respectively. Limits of detection and quantification were 0.130 and 0.391 μg/mL, respectively. High recovery value, 101.3 and low standard deviation value, 4.86 proved the accuracy of the proposed method. HPLC was used as an reference method. The method was updated from British Pharmacopeia for more less analysis time. Regression equation and correlation coefficient were obtained as y=65012C-7858 and R=0.9994, respectively. High recovery value, 97.16% and low standard deviation value, 4.52, proved the accuracy of the HPLC method. Spectroflourimetric method and HPLC method were compared by applying Student’t and Fisher Tests to both methods. As a result, it can be said that accuracy and precision of the proposed method are very high.