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Assay Development for Aminoglycosides by HPLC with Direct UV Detection
Bart Blanchaert1, Shengyun Huang1, Katarzyna Wach1
1Department of Pharmaceutical and Pharmacological Sciences, Pharmaceutical Analysis, KU Leuven - University of Leuven, Herestraat 49, B-3000 Leuven, Belgium.
Journal of Chromatographic Science
|April 5, 2017
Summary
Direct UV detection of aminoglycosides like tobramycin and amikacin is now possible. A novel analytical method using borate in the mobile phase enables accurate quantification without derivatization.
Area of Science:
- Analytical Chemistry
- Pharmaceutical Analysis
Background:
- Aminoglycosides lack UV chromophores, complicating direct analytical method development.
- Borate complexation can facilitate UV detection of compounds with vicinal diols.
Purpose of the Study:
- To develop and validate a direct UV detection assay for tobramycin, an aminoglycoside lacking vicinal diols.
- To extend the assay to amikacin, another therapeutically important aminoglycoside.
Main Methods:
- High-Performance Liquid Chromatography (HPLC) using an XBridge C18 column.
- Mobile phase: methanol-disodium tetraborate buffer (0.1 M, pH 9.0)-water (20:20:60) with sodium octanesulfonate.
- Isocratic elution at 40°C column temperature.
Main Results:
- A validated HPLC method for direct UV detection of tobramycin and amikacin was successfully developed.
- The method demonstrated speed, accuracy, ease of use, and cost-effectiveness.
- The assay was successfully applied to pharmaceutical formulations Amukin® and Tobrex®.
Conclusions:
- Direct UV detection of aminoglycosides, including those without vicinal diols, is feasible using borate-containing mobile phases.
- This method offers a simplified, efficient, and economical approach for routine pharmaceutical analysis of these drugs.