Method Optimization for Separation of Active Pharmaceutical Ingredients in Common Over-the-Counter Allergy Tablets by High Performance Liquid Chromatography
Start Date
23-4-2026 12:00 AM
Description
High Performance Liquid Chromatography (HPLC) is an instrumental technique that separates a mixture into individual components. There are 5 main parts to this instrument: the mobile phase, injector, pump, column, and detector. The mobile phase is one or more polar liquids in which a sample will be injected and dissolved via the injector. This solution is then pumped through a column containing a nonpolar solid, which then separates the components of the sample based on varying polarities. After the analytes are separated, the detector will indicate when a component leaves the column, and a chromatogram is produced. This research aimed to develop a procedure for instrumental analysis to be used in an educational setting. A procedure for the separation of the active ingredients in an allergy medication, using mobile phases of water and methanol, was adapted to be usable on the instrumentation possessed by the university. Further study was conducted to improve the quality of the resulting chromatograms.
Research Highlights
The Problem: Researcher Justin Malawey addressed the need to adapt an existing Waters Instruments UPLC procedure for separating active pharmaceutical ingredients in allergy medication to function on standard HPLC equipment possessed by the university.
The Method: The study employed reversed-phase gradient elution HPLC with a nonpolar solid stationary phase and a polar mobile phase consisting of methanol with 0.2% ammonia and a 10 mM ammonium acetate water buffer.
Quantitative Finding: The optimized instrument settings included a flow rate of 1 mL/min; detection wavelengths of 215 nm or 220 nm; a sample injection volume of 15 µL; and a final sample dilution of 50 ml at a 90:10 water-to-methanol ratio.
Qualitative Finding: Increasing the detection wavelength from 215 nm to 220 nm improved baseline consistency; maintaining a low methanol ratio for the initial two minutes of the run facilitated faster elution of specific analytes; detected extra peaks at 220 nm were attributed to solvent contamination rather than active ingredients.
Recommended Citation
Malawey, Justin, "Method Optimization for Separation of Active Pharmaceutical Ingredients in Common Over-the-Counter Allergy Tablets by High Performance Liquid Chromatography" (2026). 2026 Student Academic Showcase. 2.
https://digitalcommons.lindenwood.edu/src_2026/Posters/1/2
Method Optimization for Separation of Active Pharmaceutical Ingredients in Common Over-the-Counter Allergy Tablets by High Performance Liquid Chromatography
High Performance Liquid Chromatography (HPLC) is an instrumental technique that separates a mixture into individual components. There are 5 main parts to this instrument: the mobile phase, injector, pump, column, and detector. The mobile phase is one or more polar liquids in which a sample will be injected and dissolved via the injector. This solution is then pumped through a column containing a nonpolar solid, which then separates the components of the sample based on varying polarities. After the analytes are separated, the detector will indicate when a component leaves the column, and a chromatogram is produced. This research aimed to develop a procedure for instrumental analysis to be used in an educational setting. A procedure for the separation of the active ingredients in an allergy medication, using mobile phases of water and methanol, was adapted to be usable on the instrumentation possessed by the university. Further study was conducted to improve the quality of the resulting chromatograms.