Cross-Signal Contribution as a Challenge in LC-MS/MS Bioanalysis.

Area of Science:

• Analytical Chemistry
• Biochemistry
• Pharmacology

Background:

• Liquid chromatography-tandem mass spectrometry (LC-MS/MS) is a key technique for quantitative bioanalysis of small molecules.
• Despite its sensitivity and reliability, LC-MS/MS assays can be affected by interferences, particularly cross-signal contribution.
• Cross-signal contribution is an often-overlooked issue that can compromise the accuracy of bioanalytical results.

Purpose of the Study:

• To highlight the causes and impact of cross-signal contribution in LC-MS/MS bioanalysis.
• To provide practical guidance for identifying and resolving interferences in LC-MS/MS assays.
• To offer solutions for mitigating the negative effects of cross-signal contribution on quantitative results.

Main Methods:

• Analysis of seven real-world case studies involving LC-MS/MS interferences.
• Identification of various origins of cross-signal contribution.
• Development of a flowchart to systematically diagnose unexpected peaks in LC-MS/MS assays.

Main Results:

• Demonstrated diverse sources of cross-signal contribution in LC-MS/MS.
• Presented successful strategies for identifying and resolving interference-related problems.
• Illustrated the significant impact of cross-signal contribution on quantitative bioanalysis.

Conclusions:

• Cross-signal contribution is a critical challenge in LC-MS/MS bioanalysis that requires careful attention.
• Systematic troubleshooting using the provided flowchart can effectively identify and address interferences.
• Implementing appropriate measures is essential for ensuring the reliability and accuracy of quantitative bioanalytical data.