Blank Chocolate Spiked with Six Cannabinoids (0.2 mg/g) on Raptor ARC-18

	Peaks	t_R (min)	Conc. (µg/mL)*
1.	Cannabidiolic acid (CBDA)	2.18	3.33
2.	Cannabigerol (CBG)	2.44	3.33
3.	Cannabidiol (CBD)	2.58	3.33
4.	Cannabinol (CBN)	3.83	3.33
5.	Delta-9-Tetrahydrocannabinol (Δ9-THC)	4.83	3.33
6.	Tetrahydrocannabinolic acid A (THCA-A)	6.42	3.33

*Extract from a chocolate sample initially spiked at 0.2 mg/g.

Column

Raptor ARC-18 (cat.# 9314A62)

Dimensions:

150 mm x 2.1 mm ID

Particle Size:

2.7 µm

Pore Size:

90 Å

Guard Column:

Raptor ARC-18 5 mm, 2.1 mm ID, 2.7 µm (cat.# 9314A0252)

Temp.:

30 °C

Standard/Sample

Cannabinoids standard (cat.# 34014)

Cannabigerol (cat.# 34091)

d9-Tetrahydrocannabinol (cat.# 34067)

d9-Tetrahydrocannabinolic acid A (cat.# 34111)

Diluent:

75:25 Acetonitrile:water

Conc.:

Expected concentration of 3.33 ppm in final extract from chocolate initially spiked at 0.2 mg/g.

Inj. Vol.:

2 µL

Mobile Phase

Water, 5 mM ammonium formate, 0.1% formic acid

Acetonitrile, 0.1% formic acid

Time (min)	Flow (mL/min)	%A	%B
0.00	0.4	25	75
10.00	0.4	25	75

Detector	UV/Vis @ 228 nm
Instrument	UHPLC
Sample Preparation	Chocolate was pulverized using a SPEX Freezer/Mill grinder, and a 0.5 g sample was fortified with cannabinoids at 0.2 mg/g. Then, 0.5 mL of isopropyl alcohol was added to the sample. The sample was vortexed for 10 sec or until a homogenous mixture was obtained. Afterwards, 2.5 mL of acetonitrile acidified with acetic acid at 1% v/v was added to the vial. Once again, the mixture was vortexed for 30 sec, and then centrifuged for 5 min at 4300 ×g at room temperature. 100 µL of chocolate extract was mixed with 900 µL of 75:25 acetonitrile:water, and the final mix was centrifuged for 10 min at 4 ⁰C. 2 µL of final extract was injected into the HPLC-UV system.

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