Challenges in development of robust analytical methods for the quantitation of low molecular weight compounds by LC-MS/MS

Method development can be considered one of the most challenging task in a Bioanalytical laboratory. When working with LC-MS/MS and low molecular weight drugs, low degree of ionization, poor fragmentation, higher presence of isobaric compounds or analyte loss due to their high volatility can get in the way of a successful method development.

In the work we presented during the 7th Young Scientist Symposium, we aim to provide ideas and strategies to help tackle and solve these problems, based on real issues encountered during method development.

The advice given in the poster when working with low mass compounds focuses on three main points:
• Evaluate alternative ionization sources in order to potentially reduce background noise and increase overall sensitivity.
• Carefully evaluate and investigate the evaporation step in order to reduce cross-contamination, even avoid it altogether when possible.
• Higher presence of endogenous isobaric compounds, that can have the same MRM transition, means that the chromatographic separation has to be extensively optimized. Good chromatographic separation of all isobaric compounds is paramount to avoid erroneous conclusions.

You may find the poster below for your ready reference:


02/12/2021 14:33

Challenges in the Detection of Metabolic Biomarkers Using a Multi Plex Assay

A 5 Plex method for the detection of metabolic biomarkers in human plasma was developed using the U-PLEX technology on the MSD platform.

The complexities surrounding this method were the establishment of endogenous QCs for each chosen biomarker and evaluating the potential use and need for buffer QCs vs matrix QCs where applicable. Additional challenges were faced while trying to ascertain a single minimum required dilution to enable accurate quantification of all biomarkers simultaneously, encompassing the different detection ranges and limits for each biomarker.

02/12/2021 10:27

Considerations to properly assess drug stability within biological samples

Assessing drug stability during method development and validation is of paramount importance. The concentration of the target analyte must remain unaffected throughout the lifecycle of the samples to ensure the reliability of the assay data. However, a decrease in analyte concentration in a biological fluid is not always due to a lack of stability.

04/06/2021 11:22

Qualification of a method for the measurement of a potentially therapeutic protein

A recent project completed by Anapharm involved qualifying an analytical method for the measurement of a therapeutic protein in human skin tissue. The protein of interest is one that is expressed ubiquitously in a number of tissues and cell types.