Simultaneous Determination of Lyso-Gb1 and Lyso-Gb3 in Plasma Using Salt-Assisted Liquid-Liquid Extraction Combined...

Area of Science:

• Biomarker Analysis
• Analytical Chemistry
• Mass Spectrometry

Background:

• Lyso-Gb1 and Lyso-Gb3 are crucial biomarkers for Gaucher type 1 and Fabry diseases.
• Conventional quantification methods are resource-intensive, involving large solvent volumes and hazardous waste.
• There is a need for efficient, environmentally friendly diagnostic methods.

Purpose of the Study:

• To develop and validate a rapid, sensitive, and green LC-MS/MS method for simultaneous Lyso-Gb1 and Lyso-Gb3 determination.
• To utilize salt-assisted liquid-liquid extraction (SALLE) for improved sample preparation.
• To assess the clinical applicability of the method in patient samples.

Main Methods:

• Optimization of SALLE parameters including solvent and salt type/concentration.
• Method validation according to ICH M10 guidelines for selectivity, linearity, accuracy, precision, and stability.
• Application of the validated method to analyze plasma samples from healthy individuals and patients with Gaucher type 1 or Fabry disease.

Main Results:

• The SALLE-LC-MS/MS method demonstrated excellent linearity, low limits of quantification, high recovery, and minimal matrix effects for both biomarkers.
• Clinical sample analysis revealed significantly elevated Lyso-Gb1 and Lyso-Gb3 levels in respective patient groups compared to healthy controls.
• Receiver operating characteristic (ROC) analysis showed perfect diagnostic discrimination (AUC = 1.000) with 100% sensitivity and specificity for both biomarkers.

Conclusions:

• The developed SALLE-LC-MS/MS workflow is robust, high-throughput, and adheres to green chemistry principles.
• The method significantly reduces organic solvent use and hazardous waste generation.
• Its analytical performance and perfect clinical discrimination support routine use for diagnosing and monitoring Gaucher type 1 and Fabry diseases.