🚀 go-pugleaf

longlee@geocities.com wrote:
> [snipped]
> 10^9/ml. First we mix the platelet with arachidonic acid, then
> incubated for a while, the extract the products with ethyl acetate,
> evaporate the ethyl acetate, and dissolve them in methanol, and then
> apply them to the ODS-column. But we can't obtain anything. I seemed the
> reaction has not occured. I don't know why. Can someone help me how to
> control the reaction and how to isolate the products?

From what I remember when doing such work (by now 10 yr ago...) we methyl-
esterified the metabolites with diazo-methane before injection on C18 HPLC.
Also, for cleanup we used C18 cartridges. Worked perfectly and fast.

I might still be able to dig up some references if you want.

Regards,

Remco.

N.B. diazomethane is _not_ for beginners, not even in dilute solution:
carcinogen (methylates DNA just fine) and explosive when dry
(at the lab at that time, someone was synthesising the stuff, purifying it 
by distillation. They use special glassware for this: small scale, no ground
joints, use exclusively for this purpose to avoid scratches. Even though
the person doing the work was experienced, it exploded. No more front in the
fume cupboard. No personal accidents, because the guy stayed beside the cupboard
to avoid shrapnel, just in case....)