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Rapid Quantification of Yeast Lipid using Microwave-Assisted Total Lipid Extraction and HPLC-CAD

Sakda Khoomrung (Institutionen för kemi- och bioteknik, Systembiologi) ; Pramote Chumnanpuen (Institutionen för kemi- och bioteknik, Systembiologi) ; Suwanee Jansa-Ard (Institutionen för kemi- och bioteknik, Systembiologi) ; Marcus Ståhlman ; Intawat Nookaew (Institutionen för kemi- och bioteknik, Systembiologi) ; Jan Borén ; Jens B. Nielsen (Institutionen för kemi- och bioteknik, Systembiologi)
Analytical Chemistry (0003-2700). Vol. 85 (2013), 10, p. 4912-4919.
[Artikel, refereegranskad vetenskaplig]

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).



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Denna post skapades 2013-07-31. Senast ändrad 2014-10-27.
CPL Pubid: 180372

 

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Institutioner (Chalmers)

Institutionen för kemi- och bioteknik, Systembiologi (2008-2014)
Wallenberglaboratoriet (GU)

Ämnesområden

Livsvetenskaper
Analytisk kemi

Chalmers infrastruktur