Development of the first universal mixture for use in system suitability tests for High-Performance Thin Layer...

Area of Science:

• Analytical Chemistry
• Chromatography

Background:

• System Suitability Tests (SST) are crucial for routine chromatographic analysis.
• Current SST methods in HPTLC may be sample-dependent.
• A need exists for a universal, sample-independent SST approach in HPTLC.

Purpose of the Study:

• To introduce and validate a novel Universal HPTLC Mix (UHM) for HPTLC System Suitability Tests.
• To establish a sample-independent method for qualifying the entire HPTLC plate's chromatographic performance.
• To assess the UHM's suitability for detecting variations in chromatographic conditions.

Main Methods:

• Development of the Universal HPTLC Mix (UHM) comprising eight reference substances with diverse polarities.
• Evaluation of UHM chromatographic behavior across 20 different mobile phases on Silica gel 60 F254.
• Collaborative trials involving four laboratories to assess reproducibility of Rf values.
• Assessment of UHM response to variations in chromatographic parameters like humidity and mobile phase preparation.

Main Results:

• The UHM demonstrated baseline separation of at least three constituents across various mobile phases.
• Reproducibility of Rf values in collaborative trials was within a confidence interval of 0.040 Rf units.
• The UHM showed significant responses to changes in chromatographic conditions, indicating sensitivity to method variations.

Conclusions:

• The Universal HPTLC Mix (UHM) provides a reliable and sample-independent approach for HPTLC System Suitability Tests.
• The UHM effectively qualifies the entire HPTLC plate, enhancing analytical method robustness.
• This novel UHM is suitable for routine SST in HPTLC, improving data quality and reliability.