GNPS - Brittonodoxa subpinnata non-polar extract

For GC-MS analysis, plant material was extracted as following: 50 mg of frozen material was crushed and extracted with 700 uL of pre-cooled methanol (-20oC). Were added 60 uL of adonitol (0.2 mg mL-1, internal standard), with the mixture being centrifuged (10 min at 11000 g) and the supernatant transferred to glasses tubes to add chloroform (375 uL at -20oC) and ultrapure water (750 uL at 4oC), with the polar and non-polar phases being collected separately. The non-polar phase was derivatized with 50 uL of pyridine and 50 uL of BSTFA (N,O-bis-(trimethylsilyl)-trifluoroacetamide) for 1 h at 70oC; 5 uL of tridecanoic acid (2 mg mL-1) was used as internal standard.
The non-polar phase was analyzed by GC-MS equipped with an HP5-MS capillary column (30 m, 0.25 mm, 0.25 um). An initial column temperature was adjusted to 100oC for 5 min, and increasing at a rate of 5oC min-1 to a final temperature of 320oC, with a total run time of 49 min. 1 uL of injection volume with helium as a carrier gas. In both analyses a blank was injected in each 15 samples.