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Selection of high-performance liquid chromatographic methods in pharmaceutical analysis. III. Method validation.
G Szepesi1, M Gazdag, K Mihályfi
1Chemical Works of Gedeon Richter, Ltd., Budapest Hungary.
Journal of Chromatography
|March 3, 1989
Summary
This study details crucial high-performance liquid chromatography (HPLC) method validation steps, including system suitability and peak purity assessment for bisquaternary amino steroids. It introduces the absorbance-ratio method for resolving closely eluting peaks under stress conditions.
Area of Science:
- Analytical Chemistry
- Chromatography
Background:
- Method validation is critical for reliable analytical results.
- High-performance liquid chromatography (HPLC) is a widely used separation technique.
Purpose of the Study:
- To discuss essential steps in HPLC method validation.
- To demonstrate system suitability and peak purity assessment.
- To highlight methods for assessing peak resolution and system performance.
Main Methods:
- System suitability data establishment.
- Peak purity assessment using the absorbance-ratio method.
- Analysis of sample components under stress conditions.
- Characterization of separation power using system resolution (SR) and system selectivity (SS).
Main Results:
- The absorbance-ratio method effectively identifies incomplete resolution of adjacent peaks.
- System resolution and system selectivity provide metrics for HPLC system performance.
- Demonstrated application to bisquaternary amino steroids.
Conclusions:
- Robust HPLC method validation requires careful attention to system suitability and peak purity.
- The absorbance-ratio method is valuable for assessing peak resolution under stress.
- Stability-indicating methods have specific validation requirements.