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Use of high-pH (basic/alkaline) mobile phases for LC-MS or LC-MS/MS bioanalysis
1Nucro-Technics, Scarborough, ON, Canada.
Biomedical Chromatography : BMC
|October 14, 2018
Summary
High-pH mobile phases are increasingly used in liquid chromatography-mass spectrometry (LC-MS) bioanalysis, overcoming previous concerns about column stability and sensitivity. This shift enhances chromatographic performance and detection for various compounds.
Area of Science:
- Analytical Chemistry
- Bioanalysis
- Chromatography
Background:
- Traditionally, high-pH mobile phases were avoided in LC-MS/MS bioanalysis due to concerns over column instability and reduced sensitivity for basic analytes.
- These concerns stemmed from charge neutralization of basic compounds at high pH, leading to poor chromatographic behavior.
Purpose of the Study:
- To review the benefits and practical considerations of employing high-pH mobile phases in LC-MS and LC-MS/MS bioanalysis.
- To highlight improvements in chromatographic peak shape, retention, selectivity, resolution, and detection sensitivity.
- To discuss applications, cautions, and future trends for high-pH mobile phase usage in quantification.
Main Methods:
- Review of scientific literature and advancements in LC column technology.
- Analysis of the 'wrong-way-round' phenomena and its implications.
- Focus on practical aspects and examples related to quantification in bioanalysis.
Main Results:
- Advancements in LC column technology have mitigated concerns regarding column instability at high pH.
- High-pH mobile phases demonstrably improve chromatographic performance and detection sensitivity for a wide range of compounds, not just basic ones.
- The 'wrong-way-round' phenomena are now widely recognized, supporting the utility of high-pH methods.
Conclusions:
- High-pH mobile phases are a valuable and increasingly adopted tool in LC-MS/MS bioanalysis.
- Their use offers significant advantages in chromatographic separation and analyte detection, particularly for quantification.
- Further exploration of high-pH mobile phases is warranted, with ongoing advancements expected.
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