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Commission Implementing Regulation (EU) No 809/2014Show full title
Commission Implementing Regulation (EU) No 809/2014 of 17 July 2014 laying down rules for the application of Regulation (EU) No 1306/2013 of the European Parliament and of the Council with regard to the integrated administration and control system, rural development measures and cross compliance
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3. Determination of THC content
3.1. Preparation of the test sample
Stems and seeds over 2 mm in size shall be removed from the dried samples.
The dried samples shall be grinded to obtain a semi-fine powder (passing through a 1 mm mesh sieve).
The powder may be stored for 10 weeks at below 25 °C in a dark, dry place.
3.2. Reagents and extraction solution
Reagents
Δ9-tetrahydrocannabinol, pure for chromatographic purposes,
Squalane, pure for chromatographic purposes, as an internal standard.
Extraction solution
35 mg of squalane per 100 ml hexane.
3.3. Extraction of Δ9-THC
100 mg of the powdered test sample shall be weighed, be placed in a centrifuge tube and 5 ml of extraction solution shall be added containing the internal standard.
The sample shall be placed in an ultrasound bath and be left for 20 minutes. It shall be centrifuged for five minutes at 3 000 r.p.m. and then the supernatant THC solution shall be removed. The solution shall be injected into the chromatograph and a quantitative analysis shall be carried out.
3.4. Gas chromatography
(a)Apparatus
gas chromatograph with a flame ionisation detector and a split/splitless injector,
column allowing good separation of cannabinoids, for example a glass capillary column 25 m long and 0,22 mm in diameter impregnated with a 5 % non-polar phenyl-methyl-siloxane phase.
(b)Calibration ranges
At least three points for procedure A and five points for procedure B, including points 0.04 and 0.50 mg/ml Δ9-THC in extraction solution.
(c)Experimental conditions
The following conditions are given as an example for the column referred to in (a):
oven temperature 260 °C
injector temperature 300 °C
detector temperature 300 °C
(d)Volume injected: 1 μl
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