Currently working on a USP assay method for HPLC that requires water-saturated butyl chloride as a mobile phase and water-saturated chloroform as a diluent.
What is the whole point of using water-saturated solutions like this?
Would there be alternative ways to substitute these solutions?
The mechanism of any chromatography method is the selective retention of specific molecules. We use many different kinds of solid and liquid phases to effect separations different gases and liquids depending on their structure and chemical properties.
You didn’t mention what particular assay you were doing, but the use of additives to the water is to change the relative solution equilibria between the solid phase, liquid phase, and analytes. Without them you would not get the desired separation of analyte peaks for analysis.
Alternative solution blends are certainly possible. Check the literature first. Otherwise, start with the solubility characteristics of the existing solution and look for something comparable with another solvent.
Using water-saturated butyl chloride as a mobile phase and water-saturated chloroform as a diluent in HPLC maybe to enhance sensitivity, aid in phase separation, or reduce ionization. The specific reason for using such aqueous form of solution could differ based the type of assay and the APIs or analyte being analyzed. Alternatives can be considered, but method validation (still dependent on the analyte involved) is essential to ensure the reliability of results.
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