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Haifa Tawfeeq Abu Tbeekh

VALIDATION AND DETERMINATION OF
CLARITHROMYCIN AND METRONIDAZOLE IN RAT
PLASMA BY USING HIGH PERFORMANCE LIQUID
CHROMATOGRAPHY/MASS SPECTROMETRY
(HPLC/MS) IN PRESENCE OF POMEGRANATE JUICE
By
Haifa Tawfeeq Abu Tbeekh
A Thesis Submitted in Partial Fulfillment of the Requirements for the
Degree of Master of Science in Pharmaceutical Sciences at
University of Petra
Faculty of Pharmacy and Medical Sciences
Amman-Jordan
October 2013
Supervisor
Dr. Eyad Mallah
Co-Supervisor
Dr. Wael Abu Dayyih
 
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Abstract
Pomegranate juice has been widely consumed around the world especially in the Middle Eastern countries from the standpoint of its prevention and treatment of common diseases. Clarithromycin and Metronidazole are widely and concomitantly used drugs. In addition, both of them are considered as CYP3A4 metabolic enzyme substrates and inhibitors. A simple, reproducible and rapid analytical method by using high performance liquid chromatography-mass spectrometry (HPLC/MS) with high resolution and sensitivity for simultaneous quantification of Clarithromycin and Metronidazole in presence of pomegranate juice was validated. A gradient mobile phase system consisting of methanol and 0.1% of formic acid and ACE 5 C18 column (50 X 2.1 mm, 5μ) were used with a flow rate of 1.0 ml/ min.
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An acceptable recovery was achieved (97.75%) and (96.0%) for Clarithromycin
and Metronidazole, respectively, with good accuracy and precision. Coefficient of
determination (R²) of the standard curves for both drugs ranged between 0.9986 and
0.9998.
According to results obtained, there was no significant effect on pharmacokinetic
parameters of Clarithromycin after pretreatment with single and multiple doses of
pomegranate fresh juice. However, there was two hours-long delay on Tmax after the
single dose of juice which is possibly with no clinically significant. Furthermore, the
changing in t½ and the elimination rate constant of Clarithromycin after single and
multiple pomegranate juice administration give an expectation that pomegranate juice
could affect the hepatic-mediated metabolism of Clarithromycin but with insignificant
manner.
Pharmacokinetic parameters of Metronidazole was not affected by single dose
administration of juice, on the other hand, multiple dose pretreatment with juice
significantly elevating the maximum plasma concentration (Cmax) (P˂0.05) and the area
under the curve (AUC) of Metronidazole (P=0.001). Since there was a very slight
changing on the t½ and the elimination rate constant, the hepatic enzymes could not be
affected. In contrast, concomitant administration of Metronidazole and Clarithromycin
showed no significant interaction.
Since there are different enteric metabolic enzymes involved in the orally
administered drug metabolism, further in vitro and in vivo investigations to study the
mechanism of the pomegranate juice effect on the pharmacokinetic parameters of these
drugs should be established.
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DEDICATIONS
To my parents, my husband, and my wonderful children.
You are my family which I always proud of. I hope you are proud of me as
well by this work.