3 research outputs found
Rapid Quantification of Yeast Lipid using Microwave-Assisted Total Lipid Extraction and HPLC-CAD
We here present simple and rapid
methods for fast screening of
yeast lipids in <i>Saccharomyces cerevisiae</i>. First we
introduced a microwave-assisted technique for fast lipid extraction
that allows the extraction of lipids within 10 min. The new method
enhances extraction rate by 27 times, while maintaining product yields
comparable to conventional methods (<i>n</i> = 14, <i>P</i> > 0.05). The recovery (<i>n</i> = 3) from
spiking
of synthetic standards were 92 ± 6% for cholesterol, 95 ±
4% for triacylglycerol, and 92 ± 4% for free fatty acids. Additionally,
the new extraction method combines cell disruption and extraction
in one step, and the approach, therefore, not only greatly simplifies
sample handling but also reduces analysis time and minimizes sample
loss during sample preparation. Second, we developed a chromatographic
separation that allowed separation of neutral and polar lipids from
the extracted samples within a single run. The separation was performed
based on a three gradient solvent system combined with hydrophilic
interaction liquid chromatography-HPLC followed by detection using
a charged aerosol detector. The method was shown to be highly reproducible
in terms of retention time of the analytes (intraday; 0.002–0.034%
RSD; <i>n</i> = 10, interday; 0.04–1.35% RSD; <i>n</i> = 5) and peak area (intraday; 0.63–6% RSD; <i>n</i> = 10, interday; 4–12% RSD; <i>n</i> =
5)