Method 

reference for the presence of Cannabidiol (CBD) from HSA Singapore

Sample Preparation GCMS Conditions:

1.Weigh 0.45 – 0.55 g of sample into a 15-ml Column

polypropylene tube and add 4 ml of 0.2 N
Rtx® - 5Sil MS or equivalent
methanolic potassium hydroxide (by
dissolving 1.12 g of potassium hydroxide in 5% diphenyl / 95% dimethyl polysiloxane
100 ml of methanol.
30 m, 0.25 mm ID, 0.25 μm
2.Vortex the mixture to disperse, followed by
washing with 3 x 4 mL of n-hexane, with the MS Ionization mode
upper organic layer discarded to waste. Electron Impact (EI) at 70 eV
Note: If phase separation is not distinct, Solvent delay
centrifuge for about 3 mins at 1500 rpm
before discarding the upper organic layer 5 min

3.Carry out another wash with 4 mL of n- Inlet Mode

hexane and 1 mL of water. Split (20:1 ratio)
4.Discard the upper organic layer and add 1.5 Inlet Temperature
mL of 1 M hydrochloric acid. (by diluting 85 ml
of hydrochloric acid with water to 1000 ml). 250oC
Mix the solution.
Transfer line Temperature
5.Add 3 mL of n-hexane : ethyl acetate (9:1,
250oC
v/v) to the solution. Mix and centrifuge for
about 5 min at 1500 rpm. Repeat another Ion Source Temperature
round of centrifuge if phase separation is not
230oC
distinct.
MS Quad Temperature
6.Transfer upper organic layer to a glass test
tube and evaporate to dryness in solvent 150oC
concentrator with nitrogen gas.
Carrier flow rate
7.Add 5 ml of ethanol into the test tube and
1.00 mL/min
vortex well to disperse.
Injection Volume
8.Filter the mixture through 0.20 μm PTFE
membrane filter (Sample Solution). 1 µl

Scan Range
Sample Blank 50 to 400
1. Sample Blank Solution: Carry out the
procedure as given above without sample.
Method reference for the presence of Cannabidiol (CBD) from HSA Singapore

Oven temperature programming:

Name

Oven ramp Mass

Rate (oC/min) Q1

Next Temperature (oC) Cannabidiol

Hold (CBD)

(min) 314.4636

Runtime 315.1

(min) Cannabinol

Initial (CBN)

- 310.4319

200 311.2

2 ∆9 – Tetrahydrocannabinol

2.0 (THC)

Ramp 1 314.4690

10 315.2

240

8.0

Ramp 2

10

280

18.0