Understanding the Impact of Omega-3 Rich Diet on the Gut Microbiota. - PubMed - NCBI:
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Wednesday, April 13, 2016
Saturday, April 9, 2016
simplified procedure for semi-targeted lipidomic analysis of oxidized phosphatidylcholines induced by UVA irradiation:
Combined samples from two identically treated wells in 2 ml methanol/acetic acid (3%)/BHT (0.01%) were supplemented with internal standard (1,2-dinonanoyl-sn-glycero-3-phosphocholine [DNPC], 20 ng per sample) and transferred to a 20-ml acid-washed glass tube, which was placed into a 2-l beaker containing ice and constantly purged with argon. The top of the tube was below the edges of the argon-filled beaker, thus protecting the sample from contact with air. The samples were washed with 4 ml hexane/BHT (0.01%) that was supplied from a dispenser filled with argon. The tube was additionally purged with argon, closed with a teflon-lined screw cap, and vortexed. The hexane layer was aspirated; due to significant difference in densities of the solvents, hexane could be removed almost completely. Any potential loss of methanolic phase was corrected by the presence of internal standard. After the third wash, 4 ml chloroform/BHT (0.01%) and1.5 ml HCOOH (0.7 M) were added to the methanol phase, followed by vortexing. The lower organic phase was transferred to glass vials using a Pasteur pipette, dried under argon, and stored at −70°C until the mass spectrometry analysis.
Combined samples from two identically treated wells in 2 ml methanol/acetic acid (3%)/BHT (0.01%) were supplemented with internal standard (1,2-dinonanoyl-sn-glycero-3-phosphocholine [DNPC], 20 ng per sample) and transferred to a 20-ml acid-washed glass tube, which was placed into a 2-l beaker containing ice and constantly purged with argon. The top of the tube was below the edges of the argon-filled beaker, thus protecting the sample from contact with air. The samples were washed with 4 ml hexane/BHT (0.01%) that was supplied from a dispenser filled with argon. The tube was additionally purged with argon, closed with a teflon-lined screw cap, and vortexed. The hexane layer was aspirated; due to significant difference in densities of the solvents, hexane could be removed almost completely. Any potential loss of methanolic phase was corrected by the presence of internal standard. After the third wash, 4 ml chloroform/BHT (0.01%) and1.5 ml HCOOH (0.7 M) were added to the methanol phase, followed by vortexing. The lower organic phase was transferred to glass vials using a Pasteur pipette, dried under argon, and stored at −70°C until the mass spectrometry analysis.
Monday, March 28, 2016
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