Development and Utilization of Reversed Phase High Performance Liquid Chromatography Methods for a Series of Therapeutic Agents

Jason Olbrich and Joel Corb

Abstract

Capability to quantify drug release is vital in many areas of pharmaceutical research. Accurate, rapid and repeatable detection of pharmaceutical agents allows for comparisons between drug delivery constructs prior to in vivo studies as well as identification of therapeutic levels in samples both in vitro and in vivo. Perhaps the foremost scientific method for monitoring of therapeutics is High Performance Liquid Chromatography. This process allows for clear and simple quantification of therapeutic presence in a sample, with an inherent or induced chromophore, through the detection of bound specimen to the utilized column. Multiple drugs can be used through this process and exemplary methods are presented for the pharmaceutics; cefuroxime, clindamycin, dexamethasone, dicloxacillin, doxycycline, metronidazole, oxymetazoline, paclitaxel, tobramycin, and vancomycin. Each of these drugs is analyzed using Reversed-Phase High Performance Liquid Chromatography utilizing a hydrophobic column. Independent, repeatable methods are developed for each drug. Where necessary a pre-column derivatization is used to allow for visualization of otherwise undetectable tobramycin. In the case of each drug, a standard curve is presented to show the linearity of response for the proposed method within a range of absorbance. This work shows the ability to analyze multiple drugs with a single simple, quick, cost effective system and detection methods are evaluated for each drug with a R2 value of greater than 0.99.

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