How to partition free amino acids from proteins in plant extract
trl1 at york.ac.uk
Tue Jul 31 10:44:25 EST 2001
I am trying to quantitatively recover free amino acids away from
total soluble proteins in a plant leaf extract. The extract is made of
5-100 mg frozen leaf material homogenized in 1 mL of 80% ethanol and
then centrifuged to remove particulates. The amino acids in the
extracts will ultimately be quantified by either HPLC of PITC
derivatives, or GCMS of MTBSTFA derivatives.
I have heard of a method where such extracts are passed through a
C18 SPE column to adsorb the proteins and recover the free amino
acids in the eluate. If this works, it would be an attractive
alternative to using expensive and time-consuming low-molecular-weight
cut-off centrifugation or protein precipitation methods. However, I
can't find a protocol for such a method using C18 SPE columns (i.e.
rinse, load, elution conditions). Can anyone help?
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