Consumables and benchtop equipment
homogenizer tubes, 2 mL (BioSpec 10832)
zirconia beads, 1.0 mm (BioSpec 11079110ZX)
crimp cap vials, 12 mm x 32 mm (Thermo Scientific 200 000)
crimp caps, 11 mm red rubber (Wheaton W224211-01SP)
screw cap vials, 12 mm x 32 mm (Thermo Scientific C5000-1W)
screw caps, 11 mm red rubber (Fisherbrand 501 347)
glass vial inserts, 200 uL (ThermoFisher 500 304)
scintillation vials, 20 mL (VWR 66022-128)
P200 pipettor and pipette tips
P1000 pipettor and pipette tips
aluminum blocks (capable of fitting 12 mm crimp cap vials)
crimper and decrimper for 12 mm vials
cell disruptor homogenizer (Mini BeadBeater 96, BioSpec)
digital hot plate (HP 30A digital hot plate, Torrey Pines Scientific)
vacuum centrifuge (Savant SpeedVac SPD 2010, ThermoScientific)
EI GCMS with chromatography analysis software
Chemicals (use at least reagent grade except where noted)
cis-10-heptadecenoic acid (ISTD)
concentrated hydrochloric acid
hexane, GCMS or HPLC grade
methanol, GCMS or HPLC grade
2:1 (v/v) chloroform: methanol (Folch solution)
0.25% KCl solution in DI water
methanolic-HCl reagent (made as described in steps)
Synthetic external standard (ESTD) fatty acids will be needed for each fatty acid of interest.
A synthetic internal standard (ISTD) fatty acid will be needed to quantify compound amounts by providing estimates of the fraction of FAME products recovered and present in the GCMS injection. Choose an internal standard that does not significantly interfere with pollen fatty acid compound peaks and is not obscured by contaminants.