Rapid quantitative bioanalysis using LC-HRIM-QTOFMS with flexible sample delivery and variable traveling wave...

Area of Science:

• Analytical Chemistry
• Biochemistry
• Mass Spectrometry

Background:

• High-resolution ion mobility (HRIM) enhances bioanalytical LC-MS sensitivity and throughput.
• Conventional HRIM methods face limitations in data points per peak and working range, hindering quantification.
• This research addresses these limitations for improved small molecule and metabolite analysis.

Purpose of the Study:

• To develop rapid, quantitative bioanalytical methods using HRIM.
• To overcome the trade-offs between HRIM separation speed and data quality.
• To establish high-throughput workflows for complex biological samples.

Main Methods:

• Developed new HRIM-QTOFMS workflows for rapid quantification.
• Utilized variable traveling wave profiles to increase LC peak sampling (>15 points/peak).
• Optimized LC sample introduction with feed injection and short LC columns (<5 min/sample).

Main Results:

• Achieved rapid quantification of riboflavin and flavin nucleotides in yeast samples (<5 min/sample).
• Successfully identified and quantified phytosiderophores in plant samples (<3 min/sample).
• Demonstrated agreement with validated LC-MS/MS methods for phytosiderophore quantification.

Conclusions:

• Presents a novel combination of rapid LC, HRIM, and feed injection for quantitative bioanalysis.
• HRIM traveling wave profiles enhance LC peak sampling for high-throughput analysis.
• Enables routine analysis of small molecules in biological samples, including isomer quantification.