Unit:
Κατεύθυνση Έλεγχος Φαρμακευτικών Ενώσεων (Φαρμακευτική Ανάλυση)Library of the School of Science
Supervisors info:
Λουκάς Ιωάννης Καθηγ.(Επιβλέπων), Μ. Κουππάρης Καθηγ., Γ. Φώσκολος Καθηγ.
Original Title:
Φαρμακογεννετικός έλεγχος σε δείγματα πλάσματος εθελοντών μετά από λήψη ομεπραζόλης με χρήση μεθόδων LC-MS/MS και Real Time-PCR.
Summary:
Omeprazole belongs in the class of protons pump inhibitors a category of
medicines that acting in the stomach and decreases the acidity of his content.
In the present Master Thesis a new analytical method for determination of
omeprazole in human plasma was developed, utilizing LC-MS/MS, a highly
selective and sensitive analytical technique. Pantoprazole was used as the
internal standard. For the implementation of chromatography was used analytic
column YMC - Pack (C 8) size of particles 12 μm, diameter of resources 3 m
m and dimensions 50 x 4, 6 mm, mobile phase mix of 75% of acetonitrile and
10% of formic acid 10 mM , with a flow rate of 0,6 mL/min. Detection of the
compounds on MS/MS was performed by monitoring the MRM transitions of m/z
345,93>198,20 for omeprazole and 383,70>200,10 for pantoprazole, after
ionization of the compounds in ESI+ mode and ionization source parameter
optimization. The retention time for both compounds was 0,9 min with a 1,5 min
total analysis time for every sample.
The method was validated by 6 assay conduction (RUN1-6). Calibration curve was
linear over the range 12-1500 ng/mL for every assay, as well as for the whole
assay set. Accuracy and precision of the measurements proved to be within the
permissible limits, according to the official Guidelines. Analyte plasma
stability, as well as its stability in all used solutions, was tested and well
proved. The method was applied on pharmacokinetic evaluation of 40 healthy
volunteer plasma samples after administration of the single dose of a 20 mg
omeprazole formulation.
The above study was achieved with the use of the method of chain reaction of
polymerase in real time, RT - PCR. The method of PCR leads to augment a
section of DNA at one billion times. A molecule DNA can be strengthened so
that the final quantities that will result allow the characterization and its
usage.
The target of present work was the farmacogenetic search of omeprazole in
samples of healthy volunteers, with the use of the above methods.
Keywords:
omeprazole, LC-MS/MS, Real Time-PCR, Farmakogenetic, Human Plasma
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