The 64th American Society for Mass Spectrometry annual conference (ASMS) took place last week in San Antonio, TX. This meeting is always an important one for us. This year was no exception as our collaboration with SCIEX featured significant advances for lipidomics through our joint development of the Lipidyzer™ platform. Metabolon scientists also presented new data on the development and validation of the analytical method for our Quantose™ IGT diagnostic for impaired glucose tolerance.
The Lipidyzer Platform: A Multi-Lab Validation Project
Metabolon’s Dr. Anne Evans gave this presentation at a breakfast seminar sponsored by SCIEX. The Lipidyzer is a start-to-finish lipidomics solution that analyzes more than 1,100 lipid species across 13 lipid classes. We set out to validate the precision, sensitivity and accuracy of the platform in a cross-site project.
The goal was to have similar RSD profile to benchmark platform, specifically an average of <10% RSD for lipid species at or above 1µM at all sites.
The goal was to consistently detect hundreds of lipid species across all lipid classes at all sites.
The goal was to be accurate within +/- 20% µM of quantitative data from the benchmark platform in biological sample sets.
We were able to demonstrate that the Lipidyzer platform is reproducible and accurate across sites:
• Over 700 individual lipid species were detected with <20% RSD over multiple days and sites
• The average total assay RSD was <10% for lipid species at or above 1µM concentration in plasma
• The platform was accurate with a strong agreement (R2>0.8) of determined lipid species concentrations in multiple biological data sets and accurate to within +/- 20% of a benchmark method
High-Throughput Lipidomics Workflow for Identification and Quantification of >1,000 Lipids in 13 Different Lipid Classes
Lipidomics is a rapidly emerging field within the world of ‘omics. For human studies, plasma and serum are very attractive matrices due to ease of collection and their role in lipid transport. High individual variability in lipid levels, however, can make lipidomics experiments complicated to analyze.
While genomics and metabolomics have tackled this problem through well-established high-throughput workflows, integrated high-throughput lipidomics workflows remain unusual. Most lipidomic studies still require labor-intensive, time-consuming sample preparation, which also generates large volumes of halogenated waste.
In this poster, we present an integrated approach including extraction, analysis, identification and quantification of more than 1,000 lipids in 13 separate classes in plasma and serum.
• Automated robotic-based sample preparation for complex lipid analysis followed by analysis on the Lipidyzer platform is very comparable to the more traditional, manual Bligh-Dyer extraction method.
• Robotic extraction increases platform throughput and reduces the time needed to extract a sample set by 75%.
• This method uses five times less solvent per sample and entirely eliminates the need for halogenated solvents, which are known to be environmental toxins.
Software: Nexus Point of a Targeted Lipid Analyzer
Software is the primary interaction for a user with a mass spectrometer for both data acquisition and results generation. The challenge of any sophisticated technology is the reduction of analytical complexity so as to maintain focus on the biological problem. The intricacy of quantitative lipidomics with isobaric interference and complicated sample preparation adds another layer for consideration.
There is a need for a software-driven solution that frees the user to focus on the biological problem at hand, rather than the generation and processing of data. Metabolon and SCIEX have developed lipidomics workflow software to manage sample receiving, sample preparation protocols, mass spectrometry methods, and data processing of hundreds of MRM’s for facile, quantitative lipid analysis.
Workflow. The primary interaction of the user is to enter the study sample data.
LC-MS/MS Method for Quantitation of Seven Biomarkers in Human Plasma for the Assessment of Impaired Glucose Tolerance
Early detection of prediabetes is crucial for delaying and preventing its progress to type 2 diabetes. Impaired glucose tolerance (IGT), one of three indicators for prediabetes, is typically determined by the inconvenient and increasingly unpopular oral glucose tolerance test.
Metabolon has launched a metabolite-based test for easy assessment of IGT by successfully developing and validating a robust LC-MS/MS method for the quantitation of key seven biomarkers. This test, commercially known as Quantose IGT, has successfully assessed over 3,000 patient samples.
If you'd like more information about these presentations or any of Metabolon's products and services, please contact us.