This method seeks to determine the Δ9-tetrahydrocannabinol (hereinafter referred to as THC) content of varieties of hemp (Cannabis sativa L.) As appropriate, the method involves applying procedure A or B herein described.U.K.
The method is based on the quantitative determination of Δ9-THC by gas chromatography (GC) after extraction with a suitable solvent.
Procedure A shall be used for checks on production as provided for in Article 39 of Regulation (EC) No 73/2009 and Article 30(2)(a) of this Regulation.
Procedure B shall be used in cases as referred to in Article 39(1) of Regulation (EC) No 73/2009 and Article 40(3) of this Regulation.
Procedure A: in a standing crop of a given variety of hemp, a 30 cm part containing at least one female inflorescence of each plant selected shall be taken. Sampling shall be carried out during the period running from 20 days after the start of flowering to 10 days after the end of flowering, during the day, following a systematic pattern to ensure that the sample is representative of the field but excluding the edges of the crop.
Member States may authorise sampling to be carried out during the period from the start of flowering to 20 days after the start of flowering provided that, for each variety grown, other representative samples are taken in accordance with the first subparagraph during the period from 20 days after the start of flowering to 10 days after the end of flowering.
Procedure B: in a standing crop of a given variety of hemp, the upper third of each plant selected shall be taken. Sampling shall be carried out during the 10 days following the end of flowering, during the day, following a systematic pattern to ensure that the sample is representative of the field but excluding the edges of the crop. In the case of dioecious varieties, only female plants shall be taken.
Procedure A: the sample shall comprise parts of 50 plants per field.
Procedure B: the sample shall comprise parts of 200 plants per field.
Each sample shall be placed in a fabric or paper bag, without crushing it, and be sent to the laboratory for analysis.
The Member State may provide for a second sample to be collected for counteranalysis, if required, to be kept either by the producer or by the body responsible for the analysis.
Drying of the samples shall begin as soon as possible and, in any case, within 48 hours using any method below 70 °C.
Samples shall be dried to a constant weight and to a moisture content of between 8 % and 13 %.
After drying, the samples shall be stored without crushing them at below 25 °C in a dark place.
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.
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.
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.
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.
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.
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
Volume injected: 1 μl
The findings shall be expressed to two decimal places in grams of Δ9-THC per 100 grams of analytical sample dried to constant weight. A tolerance of 0,03 g per 100 g shall apply.
Procedure A: one determination per test sample.
However, where the result obtained is above the limit laid down in Article 39(1) of Regulation (EC) No 73/2009, a second determination shall be carried out per analysis sample and the mean value of the two determinations shall be taken as the result.
Procedure B: the result shall correspond to the mean value of two determinations per test sample.
| Regulation (EC) No 796/2004 | This Regulation | Regulation (EC) No 1120/2009 |
|---|---|---|
| Article 1 | Article 1 | |
| Article 2(1) | — | Article 2(a) |
| Article 2(1a) | Article 2(1) | |
| Article 2(1b) | — | |
| Article 2(2) | — | Article 2(c) |
| Article 2(2a) | — | Article 2(d) |
| Article 2(3) | Article 2(3) | |
| Article 2(4) | Article 2(4) | |
| Article 2(5) | Article 2(5) | |
| Article 2(6) | Article 2(6) | |
| Article 2(7) | Article 2(7) | |
| Article 2(8) | Article 2(8) | |
| Article 2(9) | Article 2(9) | |
| Article 2(10) | Article 2(10) | |
| Article 2(11) | Article 2(11) | |
| Article 2(12) | Article 2(12) | |
| Article 2(13) | Article 2(14) | |
| Article 2(14) | — | |
| Article 2(15) | Article 2(15) | |
| Article 2(16) | Article 2(16) | |
| Article 2(17) | Article 2(17) | |
| Article 2(18) | Article 2(18) | |
| Article 2(19) | Article 2(19) | |
| Article 2(20) to 2(36) | Article 2(21) to 2(37) respectively | |
| Article 2(37) | — | |
| Article 2, second last subparagraph | Article 2(38 | |
| Article 2, last subparagraph | — | |
| Article 3(1) to 3(7) | Article 3(1) to 3(7) | |
| Article 4 | Article 4 | |
| Article 5 | Article 5 | |
| Article 6 | Article 6 | |
| Article 7 | Article 7 | |
| Article 8(1) | Article 34(4) | |
| Article 8(2) | Article 34(5) | |
| Article 9, first paragraph | Article 8(1) | |
| Article 9, second paragraph | Article 8(2) | |
| Article 10 | Article 9 | |
| Article 11(1) | Article 11(1) | |
| Article 11(2), first subparagraph | Article 11(2), first subparagraph | |
| Article 11(2), second subparagraph | Article 11(2), third subparagraph | |
| Article 11(2), third subparagraph | Article 11(2), second subparagraph | |
| Article 11(3) | Article 10(2) | |
| Article 12(1) a), b), c) and d) | Article 12(1) a), b), c) and d) respectively | |
| Article 12(1) e) | — | |
| Article 12(1) f) | Article 12(1) e) | |
| Article 12(2), (3) and (4) | Article 12(2), (3) and (4) | |
| Article 13(1), 1st and 2nd subparagraphs | Article 13(1) | |
| Article 13(1) third subparagraph | — | |
| Article 13(2), (3) and (4) | — | |
| Article 13(5) | Article 13(2) | |
| Article 13(6) | — | |
| Article 13(7) | Article 13(3) | |
| Article 13(8) | Article 13(4) | |
| Article 13(9) | — | |
| Article 13(10) | Article 13(5) | |
| Article 13(11) and (12) | — | |
| Article 13(13a) | Article 13(6) | |
| Article 13(14) | Article 20(3) | |
| Article 14(1), first subparagraph | Article 13(8), first subparagraph | |
| Article 14(1), second subparagraph | — | |
| Article 14(1), third subparagraph | Article 13(8), second subparagraph | |
| Article 14(1), fourth subparagraph | Article 13(8), third subparagraph | |
| Article 14(1a) | Article 55(1) and (2) | |
| Article 14(2) | Article 12(5) | |
| Article 14(3) | Article 10(1) | |
| Article 14(4) | Article 13(9) | |
| Article 15 | Article 14 | |
| Article 15a | — | |
| Article 16(1), (2) and (3) | Article 16(1), (2) and (3) respectively | |
| Article 16(3), third subparagraph | Article 65(3), third subparagraph | |
| Article 16(4) | Article 16(4) | |
| Article 17 | — | |
| Article 17a | Article 17 | |
| Article 18 | Article 20 | |
| Article 19 | Article 21 | |
| Article 20 | Article 22 | |
| Article 21 | Article 23 | |
| Article 21a(1) and (2) | Article 24 | |
| Article 21a,(3) 2nd subparagraph | Article 15(1) | |
| Article 22 | Article 25 | |
| Article 23 | Article 26 | |
| Article 23a(1), first and second subparagraphs | Article 27(1) | |
| Article 23a(2) | Article 27(2) | |
| Article 24(1)(a), (b), (c), (d), (e), (g), (i), (j) and (k) | Article 28(1)(a), (b), (c), (d), e), (f), (g), (h) and (i) respectively | |
| Article 24(1)(f) and (h) | — | |
| Article 24(2) 1st subparagraph | Article 28(2) | |
| Article 24(2) 2nd subparagraph | Article 28(3) | |
| Article 26(1), (3) and (4) | Article 30(1), (3) and (4) respectively | |
| Article 26(2)(a), (b), (c), (f) and (h) | 30(2)(a), (b), (c), (g) and (h) respectively | |
| Article 26(2)(d), (e) and (g) | — | |
| Article 27(1), first subparagraph, first sentence | Article 31(1), first subparagraph | |
| Article 27(1), first subparagraph, second sentence, points (a), (b) and (c) | Article 31(2) | |
| Article 27(1), second and third subparagraphs | Article 31(1), second and third subparagraphs | |
| Article 27(3) and (4) | Article 31(3) and (4) | |
| Article 28 | Article 32 | |
| Article 29 | Article 33 | |
| Article 30(1) first and second subparagraphs, (2), (3) and (4) | Article 34(1), (2), (3) and (6) respectively | |
| Article 30(1), third subparagraph | — | |
| Article 31 | Article 37 | |
| Article 31a | Article 38 | |
| Article 31b | Article 39 | |
| Article 32 | Article 35 | |
| Article 33(1) | — | |
| Article 33(2), (3), (4) and (5) | Article 40(1), (2), (3) and (4) respectively | |
| Article 33a | — | |
| Article 33b | — | |
| Article 33c | — | |
| Article 34(1), 1st subparagraph | Article 41(1), 1st subparagraph | |
| Article 34(1), 2nd subparagraph | — | |
| Article 34(2) | Article 41(2) | |
| Article 35(1) | Article 42(1), 1st paragraph | |
| Article 35(2)(a) | Article 42(1), 2nd paragraph | |
| Article 35(2)(b), 1st paragraph, 1st to 4th indent | Article 42(2), 1st paragraph, (a) to (d) | |
| Article 35(2)(b), 2nd paragraph | Article 42(2), 2nd paragraph | |
| Article 35(2)(c), 1st and 2nd indent | Article 42(3), (a) to (b) respectively | |
| Article 36 | Article 43 | |
| Article 37 | Article 44 | |
| Article 38 | — | |
| Article 39 | Article 45 | |
| Article 40 | — | |
| Article 41, (a), (b), (c) and (d) | Article 47(1), (2), (3) and (4) | |
| Article 42 | Article 48 | |
| Article 43 | Article 49 | |
| Article 44(1), (1a) and (2) | Article 50(1), (2) and (3) respectively | |
| Article 45(1), (1a), (1b), (2), (3) and (4) | Article 51(1), (2), (3), (4), (5) and (6) respectively | |
| Article 46 | Article 52 | |
| Article 47(1), (1a), (2), (3), (4) and (5) | Article 53(1), (2), (3), (4), (5) and (6) respectively | |
| Article 48 | Article 54 | |
| Article 49(1) | Article 56(1) | |
| Article 49(2) | Article 19(1), 3rd paragraph | |
| Article 49(3) | Article 56(2) | |
| Article 50(1), (2) and (3) | Article 57(1), (2) and (3) respectively | |
| Article 50(5) | — | |
| Article 50(7) | Article 75(1) | |
| Article 51(1) | Article 58 | |
| Article 51(2a) | Article 57(2) | |
| Article 51(3) | — | |
| Article 52 | Article 59 | |
| Article 53, 1st and 2nd subparagraphs | Article 60 | |
| Article 53, 3rd and 4th subparagraphs | Article 57(2) | |
| Article 54 | Article 61 | |
| Article 54a | — | |
| Article 54b | Article 62 | |
| Article 57(1) | Article 63(1) | |
| Article 57(2) | Article 63(2) | |
| Article 57(3), first subparagraph | Article 63(3) | |
| Article 57(3), 2nd subparagraph | Article 75(1) | |
| Article 57(4) | Article 63(4) | |
| Article 58 | Article 64 | |
| Article 59 | Article 65 | |
| Article 60 | Article 66 | |
| Article 61 | Article 67 | |
| Article 62 | Article 68 | |
| Article 63 | — | |
| Article 64 | — | |
| Article 65(1), (2a), (3), (4) and (5) | Article 70(1), (2), (3), (4) and (5) respectively | |
| Article 66(1) | Article 70(8) and 71(1) | |
| Article 66(2) | Article 70(6) | |
| Article 66(2a) and (2b) | Article 71(2) and (3) respectively | |
| Article 66(3), 1st and 3rd subparagraphs | Article 71(4) | |
| Article 66(3), 2nd subparagraph | Article 70(7), first sentence | |
| Article 66(4) and (5) | Article 71(5) and (6) respectively | |
| Article 67(1) | Article 70(8) and 72(1) | |
| Article 67(2) | Article 72(2) | |
| Article 68 | Article 73 | |
| Article 69 | Article 74 | |
| Article 70 | Article 76 | |
| Article 71 | Article 77 | |
| Article 71a | Article 78 | |
| Article 71b | Article 79 | |
| Article 72 | Article 75(2) | |
| Article 73(1), (3) and (4) | Article 80(1), (2), and (3) respectively | |
| Article 73(5), (6) and (7) | — | |
| Article 73a(1), (2), (2a), (2b), (3) and (4) | Article 81(1), (2), (3), (4), (5) and (6) respectively | |
| Article 74 | Article 82 | |
| Article 75 | Article 83 | |
| Article 76 | Article 84 | |
| Article 77 | — | |
| Article 78 | Article 85 | |
| Article 80 | — | |
| Article 81 | — | |
| Annex I | Annex II |