HIGH THROUGHPUT LC/MS PLATFORM FOR LARGE SCALE SCREENING OF BIOACTIVE LIPIDS IN HUMAN PLASMA/SERUM

Significance of the topic

Accurate lipid profiling is crucial for understanding disease mechanisms and monitoring health. High throughput lipid screening enables large scale studies with improved robustness and speed.

Objectives and Study Overview

This work describes a semi quantitative platform capable of screening over two thousand bioactive lipids in human plasma and serum. It bridges untargeted and targeted lipidomics by combining comprehensive coverage with rapid routine analysis.

Methodology and Instrumentation

A hydrophilic interaction liquid chromatography separation was performed using an ACQUITY UPLC BEH Amide column coupled to a Waters Xevo TQ-XS tandem quadrupole mass spectrometer. A library of more than four thousand multiple reaction monitoring transitions was employed. Key steps include

• Protein precipitation of 25 microlitre plasma or serum aliquots
• Polarity switching acquisition for positive and negative ion modes
• Use of commercially available internal standards and calibrants

Main Results and Discussion

Chromatographic retention times were highly reproducible across 150 injections and multiple column batches. The method demonstrated linearity over more than three orders of magnitude, adequate sensitivity for endogenous lipid levels, and low carry over. Screening of approximately 450 lipids per polarity showed excellent precision with intra day biases below 15 percent. Application to a prostate cancer cohort revealed upregulation of specific lysophosphatidylethanolamine species.

Benefits and Practical Applications

The platform supports rapid hypothesis generation in lipidomics, high throughput biomarker screening, and can be adapted for precise quantification in clinical studies. Its robust performance is well suited for large population omics investigations.

Future Trends and Opportunities

Future developments may include expanded lipid libraries, integration with untargeted workflows, automated sample processing, application of artificial intelligence for data interpretation, and clinical translation for personalized diagnostics.

Conclusion

A versatile and reliable LC MS based lipid screening method has been established, offering fast analysis, broad coverage, and adaptability for targeted follow up. This approach enhances the capacity for large scale lipidomic investigations.

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