Standards Analytical purity (> or = 97.5%) --> i.e.

, no purification needed after purchase

Stored @ 4 degrees Celsius in dark --> no thermal or photoreactions are possible
... fancy way of saying it ain't goin' nowhere.
All: 20+-1 mg in 100 mL MeOH, each
Standard solutions prepared through dilutions in MeOH
Stored as above, wrapped with parafilm and aluminum foil, until used.
Oil Samples Different cultivars, i.e. types, peanut/soybean/camellia seed/sesame/flaxseed/gr
ape seed/rapeseed/sunflower (China)
Stored @ 25 degrees Celsius in dark (details of growth in separate table of Arti
cle)
Cleaned (water?), triturated (pulverized), squashed by grinding mill
Oil presser, collected in centrifuge tube
Centrifuged 4500 rpm for 5 min --> supernatant collected (last three steps to re
move shells/exostuff and get only oils)
Also got peanut and soybean oils (not seeds) from local stores to validate metho
d (GMO versions)
ALL samples were stored in darkness @ 25 degrees Celsius before use.
To use: mechanical mortar to stir each sample (15 min at 25 degrees Celsius)
Analyzed immediatly (within 3 months) by technique in triplicate
Mixed-mode SPE - (Not for standards, of course)
Oasis WCX 60 mg (3 mL) cartridges from Waters (Milford, USA) AND *for comparison
* similar cartridges from Weltech (Wuhan, China; Hicapt SAX)
Conditioning: 3 mL MeOH, then 3 mL n-hexane
1 g oil sample weighted and diluted to 10 mL with n-hexane, then percolated with
vacuum pump @ 1.0 mL/min
Washed with 3 mL 60% isopropanol/n-hexane (remove unwanted stuff)
Elution with 2 x 1.5 mL MeOH (competes with cartridge against analyte)
Desorption solution evaporated to dryness under Nitrogen
Reconstituted in 200 uL 0.01% formic acid in 10% aqueous acetonitrile
Final extracts filtered by 0.45 um organic filter
Repeat above to ALL oil samples
Accela HPLC - TSQ Quantum Ultra EMR in selected reaction monitoring mode (SRM),
using ESI in negative mode (4kV spray, -3kV scan)
Temp @ 35 degrees Celsius on Thermo Scientific C-18 column (Hypersil Gold, 100 m
m x 2.1 mm, 3.0 um)
Mobile Phase A: 0.01% aqueous formic acid
Mobile Phase B: 100% acetonitrile
Linear gradient of mobile phase's --> 90% A for 2.0 min, A was then decreased to
20% over 9.0 min, held for 3 min, then returned to 90% for 3 min to re-equilibr
ate
Flow rate was 250 uL/min
Aliquot of 5 uL sample
MS spectra obtained by above (TSQ Quantum Ultra EMR)
Capillary temp 350 C, sheath gas pressure (N2) 30 units, auxilary (N2) 5 units,
collision gas (Ar) 1.5 mTorr, scan time 0.2 s
Peak ID SRM mode and above parameters made peaks unambiguous (baseline res or greater)
Used retention time, and parent/product ion peaks compared to authentic standard
s
(Note: above parameters were modified to optimized Sensitivity and Resolution)
Xcalibur software used for data acquisition and processing
LOD and LOQ determined as min concentrations analytes in spiked blank oil sample
s (rapeseed oil) --> S/N of 3 and 10, respectively

Statistical Note: EVERYTHING done in triplicate. Even writing their paper. And t
he amount of times they sneezed. Doesn't matter. All triplicate.
Then they used a fancy @Risk 5.5.1 software to test statistical significance of
results, which were of course good enough to be published.