Development of a robust sulfo-phospho-vanillin assay for sample normalization in LC-MS-based lipidomics

Area of Science:

• Lipidomics
• Biochemistry
• Analytical Chemistry

Background:

• Lipidomics is essential for understanding cellular processes and disease.
• Variations in sample concentration can compromise lipid quantification accuracy.
• Normalization is critical for reliable comparative lipidome studies.

Purpose of the Study:

• To introduce a robust normalization method for lipidome analysis.
• To leverage an improved sulfo-phospho-vanillin (SPV) analysis technique.
• To enhance accuracy in comparative lipid quantification.

Main Methods:

• Development of an enhanced sulfo-phospho-vanillin (SPV) colorimetric assay.
• Optimization of detection limits and reduction of sample volume requirements.
• Application of the method to diverse sample types, including saliva and cellular samples.

Main Results:

• The enhanced SPV method reliably determines total lipid content.
• Normalization improved lipid feature detection and intensity consistency across samples.
• The method demonstrated effectiveness in samples with high concentration variability without analytical bias.

Conclusions:

• SPV-based normalization offers a practical solution for accurate lipidome analysis.
• The method is accessible to research laboratories with standard equipment.
• This approach enhances the reliability of comparative lipid profile studies.