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Methods


Aminopropyl Columns






Description:

After evaporating the solvent under streaming nitrogen, the sample is resuspended in 3 ml 2-propanol/chloroform solution and applied to the aminopropyl column. The column is washed with 6 ml 2-propanol/chloroform solution. Trihydroxy fatty acids and 2-hydroxy fatty acids are eluted with 9 ml diethylether/acetic acid solution. Other oxylipins are subsequently eluted with 9 ml methanol/acetic acid solution. Both fractions are dried separately under streaming nitrogen. The remaining oxylipins are re-dissolved in 0.2 ml methanol and stored under argon atmosphere at -20°C until use.

aminopropyl column:
Supelclean LC-NH 2 SPE Tubes (3 ml, SUPELCO)
2-propanol/chloroform solution:
2-propanol/chloroform (1:1, v/v)
diethylether/acetic acid solution:
diethylether/acetic acid (98:2, v/v)
methanol/acetic acid solution:
methanol/acetic acid (98:2, v/v)



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