Isomers Recognition in HPLC-MS/MS Analysis of Human Plasma Samples by Using an Ion Trap Supported by a Linear...

Area of Science:

• Analytical Chemistry
• Biochemistry

Background:

• Isomer recognition is crucial in various scientific fields.
• Traditional methods for isomer analysis can be complex and time-consuming.
• Tandem mass spectrometry (MS/MS) offers potential for isomer differentiation.

Purpose of the Study:

• To develop and validate a rapid liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) method for isomer recognition.
• To optimize MS/MS parameters for effective isomer signal assignment.
• To apply the developed method for analyzing isomers in human plasma samples.

Main Methods:

• Utilized ion trap tandem mass spectrometry (IT-MS/MS) with helium as collision gas.
• Employed liquid chromatography (HPLC) without requiring baseline separation of isomers.
• Optimized MS/MS properties using post-processing data elaboration (LEDA) and adjusted excitation amplitude (ExA) and excitation time (ExT).
• Implemented a two-time segment acquisition strategy to reduce cycle time and improve chromatographic profiles.

Main Results:

• The developed HPLC-MS/MS method successfully differentiated between isomers without prior chromatographic resolution.
• Optimization of IT-MS/MS parameters, including ExA and ExT, facilitated isomer-specific fragmentation.
• The method demonstrated efficiency in analyzing spiked human plasma samples, confirming its applicability.
• The two-time segment approach effectively reduced acquisition time and maintained chromatographic integrity.

Conclusions:

• The proposed HPLC-MS/MS approach provides a simple, rapid, and effective solution for isomer recognition.
• The method overcomes limitations of traditional MS/MS techniques by enabling isomer differentiation without complete chromatographic separation.
• This technique holds promise for routine analysis of isomers in complex biological matrices like human plasma.