Seeking method for fatty acid content determination in plant seedsand seedlings
Not being a chemist, I am seeking a detailed protocol for
extraction and GC analysis of "free" fatty acids and fatty acids in
triglycerides (TGAs)in Arabidopsis seeds and seedlings (time-course
experiments). This is tolook quantitatively and qualitatively at the
amount and composition of TGAs in seeds of a mutant vs. wildtype and
their changes during germination and seedling establishment.
of the methods I have seen in the literature is detailed enough for
someone inexperienced to follow without assistance of an expert (none
available where I work). I am also concerned about the safety aspects
with all the dangerous organic chemicals involved. I would be very
grateful to any expert that may provide me with a detailed protocol
that can safely be followed.
I would also appreciate that
details are included for special glassware &equipment that is
required for the procedures. Please note that I do have access to a
Agilent 6890N GC machine with an automatic injector and a 30 m HP-5 30
m column, suitable for FAs.
Seeking method for fatty acid content determination in plantseeds and seedlings
Some commercial lit. can contain some useful info particularly the
right type of GC column to use, although I would search the
sicientific lit. eg
[Only registered users see links. ]
SGE recommend their BP21- Polyethylene Glycol (TPA treated) or a
BPX70-70% Cyanopropyl polysilphenylene-siloxane for the separation of
fatty acid methyl esters. There is also a cross-reference guide to
other manufacturer columns. Fisher UK who I work for also supply
Thermo Hypersil's Trace TR-WAX GC column for FA separation. Make sure
you get the right column for the job is the message, its a competative
field so you should be able to haggle a good price.