Graduation Date

Summer 8-14-2020

Document Type

Thesis

Degree Name

Master of Science (MS)

Programs

Pharmaceutical Sciences

First Advisor

Daryl J Murry, Pharm.D.

Second Advisor

Rongshi Li, Ph.D.

Third Advisor

Anthony T Podany, Pharm.D.

Abstract

A rapid, selective, and sensitive liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) method was developed and validated for quantitation of a novel Pyrrolomycin (MP-1) in mouse plasma. MP-1 was extracted from plasma utilizing a structural analog (PL-3) as the internal standard (IS). Analyte separation was achieved using a Waters Acquity UPLC®BEH C18 column (1.7 µm, 100 x 2.1 mm) protected with Acquity UPLC C18 guard column. Mobile phase consisted of 0.1% acetic acid in water (10%) and methanol (90%) at a total flow rate of 0.25 mL/min. The mass spectrometer was operated at unit resolution in the multiple reaction monitoring (MRM) mode, using precursor ion>product ion transitions of 324.10>168.30 m/z for MP-1 and 411.95>224.15 m/z for PL-3. The MS/MS response was linear over the concentration range from 0.2-500 ng/mL for MP-1 with a correlation coefficient (r2) of 0.988 or better. Precision (percent relative standard deviation, % RSD) and accuracy (% bias) were within the acceptable limits as per FDA guidelines. MP-1 was stable under storage and laboratory handling conditions. The validated method was successfully applied to assess the solubility, in vitro metabolism, plasma protein binding and bio-distribution studies of MP-1. Glucuronidation was found to be the major metabolic pathway using S9 fractions. The validated method was successfully applied to a pre-clinical pharmacokinetic and bio-distribution mouse study involving low volume blood and tissue samples for quantitation of MP-1.

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