posted on 2017-06-04, 00:00authored byFilomena Petruzziello, Alexandre Grand-Guillaume Perrenoud, Anita Thorimbert, Michael Fogwill, Serge Rezzi
Analytical solutions
enabling the quantification of circulating
levels of liposoluble micronutrients such as vitamins and carotenoids
are currently limited to either single or a reduced panel of analytes.
The requirement to use multiple approaches hampers the investigation
of the biological variability on a large number of samples in a time
and cost efficient manner. With the goal to develop high-throughput
and robust quantitative methods for the profiling of micronutrients
in human plasma, we introduce a novel, validated workflow for the
determination of 14 fat-soluble vitamins and carotenoids in a single
run. Automated supported liquid extraction was optimized and implemented
to simultaneously parallelize 48 samples in 1 h, and the analytes
were measured using ultrahigh-performance supercritical fluid chromatography
coupled to tandem mass spectrometry in less than 8 min. An improved
mass spectrometry interface hardware was built up to minimize the
post-decompression volume and to allow better control of the chromatographic
effluent density on its route toward and into the ion source. In addition,
a specific make-up solvent condition was developed to ensure both
analytes and matrix constituents solubility after mobile phase decompression.
The optimized interface resulted in improved spray plume stability
and conserved matrix compounds solubility leading to enhanced hyphenation
robustness while ensuring both suitable analytical repeatability and
improved the detection sensitivity. The overall developed methodology
gives recoveries within 85–115%, as well as within and between-day
coefficient of variation of 2 and 14%, respectively.