Beruflich Dokumente
Kultur Dokumente
Authors
Michelle Sexton 1, 2, Kyle Shelton 2, 3, Pam Haley 4, Mike West 5
Affiliations AB STR AC T
1 Center for the Study of Cannabis and Social Policy, Seattle, A recent cannabis use survey revealed that 60 % of cannabis
WA, USA users rely on smelling the flower to select their cannabis.
2 Phytalytics LLC, Kirkland, WA, USA Olfactory indicators in plants include volatile compounds,
3 Medicine Creek Analytics, Fife, WA, USA principally represented by the terpenoid fraction. Currently,
4 CO2 Garden Extracts, Redmond, WA, USA medicinal- and adult-use cannabis is marketed in the United
5 Green Lion Farms, Seattle, WA, USA States with relatively little differentiation between products
234 Sexton M et al. Evaluation of Cannabinoid … Planta Med 2018; 84: 234–241
literally, “Only the dose makes the poison” [27]. There have been
A B B R E V I AT I O N S few reports on production protocols and chemical composition of
CBD cannabidiol cannabis products being sold and used under regulated U. S. can-
CBDA cannabidiolic acid nabis markets. A report from the Netherlands compared extrac-
CBG cannabigerol tion products using naphtha, petroleum ether, ethanol, and olive
CBN cannabinol oil. This study showed differential extraction of terpenoids, yet it
SC‑CO2 supercritical carbon dioxide was unknown whether this was due to degradation, loss to evap-
THCA Δ-9 tetrahydrocannabinolic acid oration, and interaction with solvent components or whether they
THC Δ-9 tetrahydrocannabinol were simply not extracted [28]. In their report, the monoterpenes
myrcene and terpinolene and the bicyclic sesquiterpene β-caryo-
phyllene were retained across all extractions. A recent attempt
has been made to further optimize a SC‑CO2 extraction method
portable pens (electronic cigarettes or e-pens) or by a process with ethanol as a co-solvent and using focused ultrasound extrac-
known as “dabbing” [6]. Dabbing involves the flash vaporization tion to first “de-terpinate” the matrix prior to extraction of canna-
of the concentrate by rapidly heating the “dab” to 300–400 °C binoids [29]. This method used a lower temperature and allowed
(typically on a titanium rod) followed by inhalation of the vapor for retention of the monoterpenes.
with a specialized pipe. This dose has been reported to be around A recent survey of cannabis use reported that 61 % of respond-
40 mg/dab (of THC), and is reported to induce negative side ef- ers had ever “vaped,” while only 12 % report that it is their pre-
Sexton M et al. Evaluation of Cannabinoid … Planta Med 2018; 84: 234–241 235
Original Papers
Chemo- CBDA (mg/g) CBD (mg/g) THCA (mg/g) THC (mg/g) CBN (mg/g) CBG (mg/g)
var F C F C F C F C F C F C
Cherry 0.3 91.2 ND 5.3 264 693.8 1.6 77.5 ND 1.5 ND ND
Kush (± 0.06) (± 2.1) (± 0.6) (± 3.5) (± 52) (± 0.9) (± 1.9) (± 0.06)
Black- 0.5 7.0 ND 0.9 267.4 749 40.9 105.6 1.0 11 2.0 7.9
berry (± 0.02) (± 0.5) (± 0.6) (± 2.8) (± 84) (± 1.2) (± 2.3) (± 0.0) (± 0.6) (± 0.0) (± 0.6)
Kush
Pineap- 0.5 3.3 ND 1.9 235.6 650.5 24.3 174.3 ND 1 ND 1.1
ple Kush (± 0.03) (± 0.6) (± 0.5) (± 2.6) (± 45) (± 1.5) (± 3.8) (± 0.2) (± 0.2)
Purple 0.5 20.3 ND ND 24.24 674.9 19.4 9.98 0.1 15 1.3 20.5
Sour (± 0.02) (± 6.0) (± 3.1) (± 44) (± 1.0) (± 2.7) (± 0.01) (± 0.07) (± 0.0) (± 0.9)
Diesel
Ripped 0.4 3.1 ND ND 24.58 714.7 ND 9.68 ND 1.6 ND 11.4
Bubba (± 0.02) (± 0.2) (± 4.2) (± 53) (± 2.1) (± 0.7) (± 0.8)
Harle- 117.5 444.6 1.7 17.4 88 333 9.3 55.0 ND ND 0.7 ND
quin (± 2.1) (± 3.7) (± 0.6) (± 0.9) (± 1.4) (± 26) (± 2.2) (± 1.5) (± 0.03)
236 Sexton M et al. Evaluation of Cannabinoid … Planta Med 2018; 84: 234–241
Downloaded by: Kaohsiung Medical University. Copyrighted material.
▶ Fig. 2 Each flower with its predominant terpenoid chemotype is detailed. The terpenoid potency in the flower (black bar) is compared to the
concentrate (white bar). Asterisks indicate significant differences, and red asterisks indicate flower having a higher concentration than the SCCO2
concentrate. (*p < 0.05; **p < 0.005; ***p < 0.0005)
Sexton M et al. Evaluation of Cannabinoid … Planta Med 2018; 84: 234–241 237
Original Papers
Data are expressed as the mean in mg/g of starting material with standard deviation in parenthesis. Each sample was run in triplicate. Statistical significance
is indicated by an asterisk (p < 0.05). F: flower; C: concentrate.
Data are expressed as the mean in mg/g of starting material with standard deviations in parenthesis. Each sample was run in triplicate. Statistical significance is
indicated by an asterisk (p < 0.05). F: flower; C: concentrate.
procedures and validated methodologies for quantification of quire quality control testing for contaminants, such as pesticides,
compounds of interest have been lacking. Regardless, cannabi- which can be concentrated in extracts [37].
noid potency is routinely reported on certificates of analysis and Because this SC‑CO2 extraction protocol enhanced the potency
product labels to 2–3 decimal points, lending an inference of pre- of both cannabinoids and terpenoids in a differential fashion, fur-
cision that is may not be actual. ther studies are necessary to determine the health-related conse-
The market share for extracts in Washington State increased by quences of inhaling concentrated cannabis extracts [38, 39]. A
145.8 % between 2014 and 2016, accounting for one-fifth of all qualitative needs assessment of cannabis farm workers and own-
sales [35]. These types of products have been reported elsewhere ers found that many employers and employees identified terpe-
as having an 86 % contamination rate with multiple chemical noids as the causative agent of their occupational allergies. Aller-
agents: insecticides, miticides, fungicides, and growth regulators gies presented in the form of respiratory distress and contact der-
[36]. The historic/legal framework has led to substantial knowl- matitis. Currently, research does not identify the causative agent
edge gaps resulting in policies that fail to protect public health. of occupational allergies in cannabis grow operations, attributing
For instance, initially Washington State regulations failed to re- symptoms to generalized cannabis allergy [40].
238 Sexton M et al. Evaluation of Cannabinoid … Planta Med 2018; 84: 234–241
Bubba, and Harlequin (laboratory accession IDs were STR 1378,
1382, 1382, 1383, 1394). The flower trim was collected for ex-
traction when the dried flower was manicured, thus trim and
flower were treated identically prior to extraction. Flower material
and trim were collected and identified in May 2014 by Shawn
DeNae (Washington Bud Company), the licensed grower in Wash-
ington State. As a result of legal restrictions applied to cannabis by
the Controlled Substances Act (1970) to implement the Single
Convention on Narcotic Drugs (1961), we were unable obtain
voucher specimens from a major regional herbarium. We have in-
sured the authenticity of the specimens by obtaining them di-
rectly from the production source.
SC‑CO2 extraction: 100 g of trim was prepared by grinding to a
fine consistency (SharkNinja NJ600) thus increasing the surface
area for SC‑CO2 to contact and vaporize all of the compounds.
The powder was the packed into a 5-L extraction vessel (Eden Labs
2000 PSI SFE Hi-Flo). Cannabis was extracted in a closed-loop sys-
tem that continued for about 6 h. For this study, temperature for
▶ Fig. 3 This graph depicts potency changes of 14 terpenoids
Sexton M et al. Evaluation of Cannabinoid … Planta Med 2018; 84: 234–241 239
Original Papers
15 µg/mL prior to sample quantification (▶ Table 1). Response of pha = 5.000 %. Each row was analyzed individually, without assum-
the mass spectra was monitored by the use of a control standard ing a consistent standard deviation (SD).
using 10 µg/mL of the standards. There was a solvent delay of
12.5 min. Here we report only the 15 most abundant compounds Supporting information
identified by MS occurring well above the limit of detection in mg/ Additional data on the method are provided as Supporting Infor-
gram of starting material. The method was developed and deter- mation.
mined to be “fit for purpose” and sufficient for comparison in a
single laboratory between two matrices.
Acknowledgements
Terpenoid reference standards Dr. Darryl J. Bornhop for review of the manuscript and Kyle Shel-
Flower and concentrate were analyzed for 42 terpenoids (100 µg/ ton for method development and performing the laboratory ex-
mL in methanol) from a common stock solution (SPEX CAN- periments.
TERP‑MIX 1 and 2) and quantified using standard curves prepared
gravimetrically in DCM/M. To measure recovery, the reference Conflict of Interest
standards were spiked at low, medium, and high levels onto dried,
ground straw (0.25 g) and extracted in DCM/M. Calibrations stan- There are no conflicts of interest to divulge.
dards were stored at − 20 °C in the dark and tested at regular inter-
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