title: Rapid quantification of yeast lipid using microwave-assisted total lipid extraction and HPLC-CAD.
abstract: We here present simple and rapid methods for fast screening of yeast lipids in Saccharomyces cerevisiae. 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 (n = 14, P > 0.05). The recovery (n = 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; n = 10, interday; 0.04-1.35% RSD; n = 5) and peak area (intraday; 0.63-6% RSD; n = 10, interday; 4-12% RSD; n = 5).
authors: Systems and Synthetic Biology, Department of Chemical and Biological Engineering, Chalmers University of Technology, Göthenborg, Sweden.
authors: Sakda
authors: S
authors: Khoomrung
authors: Pramote
authors: P
authors: Chumnanpuen
authors: Suwanee
authors: S
authors: Jansa-Ard
authors: Marcus
authors: M
authors: Ståhlman
authors: Intawat
authors: I
authors: Nookaew
authors: Jan
authors: J
authors: Borén
authors: Jens
authors: J
authors: Nielsen
mesh_terms: D000336:Aerosols; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D008055:Lipids / Q000032:analysis* / Q000302:isolation & purification*; D008872:Microwaves; D015203:Reproducibility of Results; D012441:Saccharomyces cerevisiae / Q000737:chemistry*; D013997:Time Factors
chemical_list: D000336:Aerosols; D008055:Lipids