Methylation of Fatty Acids (Kropinski Method) fatty acid methylation
-
Last Update: 2020-10-31
-
Source: Internet
-
Author: User
Search more information of high quality chemicals, good prices and reliable suppliers, visit
www.echemi.com
Hancock Laboratory Methods,Department of Microbiology and Immunology,
University of British Columbia, British Columbia, Canada
OBJECTIVE:
To methylate fatty acids in whole cells or lipopolysaccharide.
REAGENTS :Methanol-Hydrochloride Reagent Kit
>10mg of whole cells or 1 mg of lipopolysaccharide
400 nmoles of fatty acid standard (pentadecanoic acids C15)
METHODS:
Prepare 1M MeOH-HCl reagent according to the instruction
given with the kit.
Add internal standard (C15 fatty acid) to give a final concentration of 400 nmoles/100ml (usually 10mg of C15 in 100ml of reagent.
Weigh 10mg of whole cells or 1 mg of LPS into a clean screw-cap tube.
Add 1ml of MeOH-HCl reagent/internal standard into each tube and vortex.
Heat at 100oC for 20 minutes. (Note: each tube should be very well sealed with teflon tape and grease to prevent evaporation.)
Sonicate and heat at 100oC overnight.
Neutralize acidity with 0.5N NaOH. Test pH with pH paper.
Centrifuge in clinical centrifuge for 5 minutes. Save supernatent in clean glass vial.
Do gas chomatography with programmed REWH method.
This article is an English version of an article which is originally in the Chinese language on echemi.com and is provided for information purposes only.
This website makes no representation or warranty of any kind, either expressed or implied, as to the accuracy, completeness ownership or reliability of
the article or any translations thereof. If you have any concerns or complaints relating to the article, please send an email, providing a detailed
description of the concern or complaint, to
service@echemi.com. A staff member will contact you within 5 working days. Once verified, infringing content
will be removed immediately.