AOAC 2018 Methods

experiment, a targeted analyte list was used (see Table 2018.11C ). Internal mass calibration was performed throughout the analytical run using a commercial calibration mixture delivered to secondary probe. LC eluent was diverted into waste from 0-0.5 and 12-16 minutes to protect the ion source from contamination.

1. Ion source:

dual Agilent Jet Stream positive electrospray

2. Ionization mode: 3. Gas temperature: 4. Drying gas flow: 5. Nebulizer pressure: 6. Sheath gas temperature: 7. Sheath gas flow: 8. Capillary voltage: 9. Nozzle voltage: 10. Fragmentor: 11. Octapole 1 RF: 12. Acquisition mode: 13. Mass range (full MS): 14. Acquisition rate (full MS): 15. Mass range (MS/MS): 16. Acquisition rate (MS/MS):

240°C

12 L/min 25 psig 12 L/min 4000 V 400°C

400 V 400 V 750 V

MS and targeted MS/MS (2GHz EDR)

m/z 100-1000 5 spectra/s m/z 50-500 5 spectra/s

17. Collision gas:

nitrogen enabled

18. Reference mass correction:

19. Reference mass 1: 20. Reference mass 2:

m/z 149.023320

m/z 922.009798 Note: Ion mobility option was switched off in all measurements.

Table 2018.11C. Targeted MS/MS list

Delta Retention

Collision Energy (V)

Precursor ion ( m/z )

Retention Time (min)

Analyte

Ion Type

Isolation Width

Time (min)

CBDVA

331.1904 287.2006 359.2217 343.2268 317.2475 315.2319 287.2006 311.2006 315.2319 315.2319 359.2217 315.2319

[ M +H] + [ M +H] + [ M +H] + [ M +H] + [ M +H] + [ M +H] + [ M +H] + [ M +H] + [ M +H] + [ M +H] + [ M +H] +

3.15 3.62 4.59 4.90 5.25 5.40 5.62 7.20 8.10 8.30 8.63 9.10

0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5

Narrow (1.3 Da) Narrow (1.3 Da) Narrow (1.3 Da) Narrow (1.3 Da) Narrow (1.3 Da) Narrow (1.3 Da) Narrow (1.3 Da) Narrow (1.3 Da) Narrow (1.3 Da) Narrow (1.3 Da) Narrow (1.3 Da) Narrow (1.3 Da)

20 20 20 20 20 20 20 20 20 20 20 20

CBDV CBDA CBGA

[ M –H 2 O+H] +

CBG CBD

THCV

CBN

Δ 9 -THC Δ 8 -THC

THCA

CBC

(d) System Suitability. —Run a full set of calibration standards at the beginning and the end of each sample set. Chromatographic resolution (R) between CBG and CBD and Δ 9 -THC and Δ 8 -THC in standards has to be ≥ 1.0. The calibration curve based on five concentration levels should have r 2 value of ≥ 0.995. The calculated concentrations of