Quantitative determination and validation method of amoxicillin in human plasma by HPLC

SUBJECT of THESIS: Quantitative Determination and Validation Method of Amoxicillin inHuman Plasma by HPLCSTUDENT NAME: Hatice Demet ÇEL KSUPERVISOR: Prof. Dr. Tuncel ÖZDENDEPARTMENT: Faculty of Pharmacy, Department of Pharmaceutical ChemistryYEAR: 2007STATUTE: MasterSUMMARYWithin the context of this study, a method was developed forquantitative determination of a β-lactam antibiotic derived from penicillin and hassignificant importance among antibacterial drugs designated by the name ofamoxicillin.Amoxicillin in human plasma was incorporated to solvent system vialiquid-liquid extraction method, and quantitative determination was performed byusing HPLC and fluorescence detector. For the purpose of quantitativedetermination, detector was adjusted to 335 nm for excitation, to 435 nm foremission and column temperature was arranged to 40 0C. Run time was 10 minuteswhilst the mobile phase flow rate was 1 ml/min. as internal standard; ampicillin wasused while implementing the method. In the scope of this study, it was proved thatthe methodology is linear between 0.1-20 µg/mL concentration interval. Relativerecovery was calculated at 77.33 %.What follows the determination of method suitability is the validationof method according to FDA rules. Throughout the validation procedure,parameters of specificity, selectivity, accuracy, precision, linearity, recovery andstability were determined and the results were evaluated statistically.After the validation studies it was determined that the developedmethodology is sensitive, reproducible and easy to practice in the contemplation ofpharmacokinetic, pharmacodynamic and bioavailability bioequivalence studies ofamoxicillin.