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Updated: Jul 16, 2026

Preparation of Human Tissues Embedded in Optimal Cutting Temperature Compound for Mass Spectrometry Analysis
Published on: April 27, 2021
Using strong injection solvents with 100% aqueous mobile phase in RP-LC
1Chemical and Analytical Development, Novartis Pharmaceuticals, East Hanover, NJ 07936, USA. eric.loeser@novartis.com
Abstract:
When samples are dissolved for HPLC analysis, organic solvents are often used to enhance the solubility of the sample components. However, when the diluent becomes significantly stronger than the mobile phase, peak shape distortions may develop during injection. This is a serious problem for weakly retained analytes under 100% aqueous mobile phase conditions. Under these extreme conditions, even small amounts of solvent in the diluent can cause serious broadening or distortion effects. However, if the organic solvent used in the diluent elutes significantly after the analyte, the peak shape of the analyte will be relatively unaffected, even when a relatively strong solvent such as THF is used. This phenomenon is counterintuitive considering the usual practice of keeping the sample diluent as weak as possible. This report describes the potential analytical utility and limitations of this approach for the analysis of weakly retained analytes.
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