Post-graduate theses
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Identifier |
000463795 |
Title |
Ανάπτυξη και επικύρωση μεθόδου ανάλυσης κανναβινοειδών σε δείγματα αμφιβληστροειδούς με τη χρήση LC-MS/MS |
Alternative Title |
Method development and validation for the analysis of cannabinoids in retina samples using LC-MS/MS |
Author
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Λαποκωνσταντάκη, Εμμανουέλα Γ.
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Thesis advisor
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Χανιωτάκης, Νικόλαος
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Reviewer
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Θερμού, Κυριακή
Χατζηιωάννου, Αναστασία-Χρυσοβαλάντου
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Abstract |
The plant Cannabis sativa has been used for medicinal purposes for many years. Τhe
endocannabinoid system (ECS) has been shown recently to be a major target for the
development of CNS (central nervous system; brain and retina) disease therapeutics, whose
pathophysiology involves neurodegeneration and neuroinflammation. The ECS system
comprises of the endocannabinoids, cannabinoid CB1 and CB2 receptors(R) and the enzymes
that synthesize and metabolize the endocannabinoids. The aim of the present master's thesis
is the employment of LC-MS/MS, methodology for quantitative analysis of experimental rat
retinal samples of a) the endocannabinoid 2-arachidonoylglycerol (2-AG) and the dual
inhibitor, AM11920, of metabolic enzymes MAGL/ABHD6 of 2-AG, after chronic
intraperitoneal administration in an excitotoxicity model and b) two synthetic analogues,
AM1710 (CB2R agonist) and SR141716 (CB1R antagonist) after topical (eye drop)
administration in an early stage diabetic retinopathy model.
The qualitative and quantitative determination was performed on a Q Exactive™ Plus
Quadrupole-Orbitrap™ Mass Spectrometer. Determination and quantification of 2-AG,
AM1710 and SR141716 was performed with an Ultimate 3000 HPLC System. An Hypersil
Gold separation column (100mm × 4.6mm i.d., 5μm) was used. The separation was performed
at a flow rate of 0.4 mL/min by gradient elution, using H2O-0,1%Formic Acid and ACN-0,1%
Formic Acid as solvents. For the mass spectrometry, electrospray ionization in positive mode
and the PRM mode for the fragmentation of the analytes were used after the optimization of the
monitoring conditions for the target compound. All methods were validated for each analyte,
in terms of linearity, precision, repeatability, robustness, limits of detection and quantification.
The use of the developed methodology of the three cannabinoids certified for the first
time that these compounds, independent of the route of administration, reached the retina and
induced their neuroprotective and anti-inflammatory actions.
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Language |
Greek |
Subject |
2AG |
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AM1710 |
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Electrospray |
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Limit of quantification |
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Limits of detection |
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Linearity |
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Liquid chromatography |
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Mass spectrometry |
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PRM |
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Precision |
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Receptors |
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Repeatability |
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Robustness |
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SR141716 |
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Όριο ανίχνευσης |
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Όριο ποσοτικοποίησης |
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Ακρίβεια |
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Ανθεκτικότητα |
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Γραμμικότητα |
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Επαναληψιμότητα |
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Ηλεκτροψεκασμός |
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Κανναβινοειδή |
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Υγρή χρωματογραφία |
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Υποδοχείς |
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Φασματομετρία μάζας |
Issue date |
2024-07-19 |
Collection
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School/Department--School of Sciences and Engineering--Department of Chemistry--Post-graduate theses
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Type of Work--Post-graduate theses
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Permanent Link |
https://elocus.lib.uoc.gr//dlib/2/3/b/metadata-dlib-1712300571-683617-3625.tkl
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Views |
464 |
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