Abstract
Microsampling devices enable the rapid collection of thousands of blood samples, making them particularly advantageous for high-throughput metabolic profiling in large-scale studies. However, their integration with a fast analytical method enhances their compatibility, improving the workflow by minimizing the sample volume, reducing analysis time, accelerating data acquisition, and decreasing the risk of technical bias compared to conventional methods.
Rapid Microbore Metabolic Profiling (RAMMP) methods were developed to provide ultra-fast liquid chromatography analysis cycles of up to five minutes while maintaining high retention time repeatability. Although RAMMP has been successfully applied to urine and plasma, its performance in blood microsampling devices or whole blood has not yet been evaluated. This study assesses the feasibility of the RAMMP method for metabolic analysis of blood microsampling devices against plasma and whole blood and compares its performance with a conventional UPLC-MS approach. Conventional untargeted reversed-phase lipid profiling was performed using a UPLC ACQUITYTM BEH C8 (2.1 × 100 mm, 1.7 μm) column with a 0.6 mL/min flow rate over 13.25 minutes and 2 μL volume injection. This method was scaled down using a UPLC ACQUITYTM BEH C8 (1.0 × 50 mm, 1.7 μm) column, with a reduced flow rate of 0.3 mL/min over 3.7 minutes and 1 μL injection volume. Blood and plasma samples from 10 healthy volunteers were provided by the HUMAN Doctoral Network. Data acquisition was conducted on a Synapt G2-S system in positive ionisation mode, with data processing performed using Metabonaut and R for mass spectrometry package. Results demonstrate that RAMMP significantly reduces analysis time while maintaining comparable chromatographic performance to conventional UPLC-MS methods. Although RAMMP exhibits lower peak capacity and detects fewer ions, it achieves similar lipid group discrimination, supporting its potential as a high-throughput alternative for large-scale microsampling-based studies.