Analysis of Residual Solvents in Hemp Extract

by Headspace SPME-GC/MS
A straightforward, fast, sensitive, and accurate alternative to headspace analyzers

and that there was no carryover between standards

and samples. A Carboxen® (CAR)/PDMS SPME fiber was
chosen for the extraction step due to its ability to retain
low molecular weight compounds with differing polarities,
such as hexane and methanol. The column for the GC/
MS analysis, Supel-Q™ PLOT, was chosen for its ability
to retain very low molecular weight compounds, and for
its selectivity toward a variety of compounds, including
alcohols, aliphatics, and aromatics. Supel-Q™ PLOT also
has the ability to retain lighter aliphatics, such as butane
and pentane, and thus would be a suitable choice if these
compounds are to be analyzed.

Figure 1. Hemp Extract Spiked at 10 µg/g with Solvents,

Analyzed by SPME-GC/MS
sample/matrix: 5 g hemp oil in 10 mL vial
SPME fiber: SPME fiber assembly Carboxen/Polydimethylsiloxane (CAR/PDMS)
(57319)
extraction: 5 min, headspace, 40 °C
desorption 3 min, 320 °C; split 10:1
process:
fiber post bake: 2 min, 320 °C
Marijuana oil, produced by extraction of the flower buds column: Supel-Q™ PLOT,
30 m × 0.32 mm I.D. (24242)
of the cannabis sativa plant, has reputed therapeutic oven: 50 °C (5 min), 10 °C/min to
benefits.1,2 The extraction process typically involves 230 °C (5 min)
inj. temp.: 320 °C
solvents such as hexane or some alcohols that may detector: MSD
remain in the final product.3 The residual solvents are MSD interface: 260 °C
typically measured using a headspace analyzer interfaced scan range: m/z 29-200
carrier gas: helium, 2 mL/min constant flow
directly to a GC. Solid phase microextraction (SPME) is injection: split, 10:1
an alternative approach that is often faster and easier to liner: SPME, 0.75 mm I.D.
instrument: Agilent® 6890/5973N GC/MS
use. The method outlined here uses SPME with a GC/MS
finish to measure solvents in hemp extract. Hemp and 1. Methanol 8. Benzene
2. Ethanol 9. Cyclohexane
marijuana are both varieties of cannabis sativa, however 3. Acetonitrile 10. Heptane
hemp is a variety that is very low in THC content. 4. Acetone 11. Toluene
5. Isopropanol 12. m & p-Xylenes
6. Tetrahydrofuran (THF) 13. o-Xylene
Experimental 7. Hexane
12
Analytical conditions appear in Figure 1. The sample 3
comprised 99% pure hemp extract in a base of hempseed 11
oil, unspiked and spiked at 10 µg/g with the solvents listed 4 7,8 9
in Table 1. Quantitation was by external standard against 2 5 6
10 13
1
a 6-point curve from 6 to 100 μg/g prepared in soybean
oil. Blank soybean oil was analyzed along with SPME fiber 4 6 8 10 12 14 16 18 20 22
blanks to verify cleanliness of the system prior to testing Min

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Results and Discussion Featured and Related Products
A representative chromatogram of a spiked hemp extract Description Cat. No.
sample is shown in Figure 1. The coelution of benzene/ Reference Materials
cyclohexane was resolved using unique m/z values for Acetonitrile PHR1320
quantitation of each compound. A summary of accuracy
Benzene PHR1318
and precision for the analyses of n=3 replicates of hemp
extract, spiked at 10 µg/g, is presented in Table 1. Methanol PHR1170
Hexane was present in the unspiked hemp extract at Toluene PHR1317
six times the spiking level, and thus, is not reported for Chloroform PHR1316
the spiked samples. The hexane level reported for the
Methylene Chloride PHR1171
unspiked extract is the average of duplicate analyses, with
the percent reproducibility of the measurement (13%) Trichloroethylene PHR1319
shown in parentheses. For the remaining solvents in the Reference Materials - Residual Solvent Mixes
analyte list, accuracy was >90% for all compounds except Residual Solvents Mixture Class I PHR1063
ethanol, which was 84%. Precision, determined as % RSD, Benzene 1,1-Dichloroethane
was good with values of <10% for all spiked compounds. Carbon tetrachloride 1,1,1-Trichloroethane
1,2-Dichloroethane
The use of SPME allowed for the straightforward,
fast, sensitive, and accurate GC/MS determination of Residual Solvents Mixture Class IIA PHR1064

the residual solvent content of spiked and unspiked Acetonitrile Methylcyclohexane

samples of hemp extract. The CAR/PDMS SPME fiber Chlorobenzene Tetrahydrofuran
in combination with the Supel-Q™ PLOT GC column trans-1,2-Dichloroethylene Toluene/acetonitrile
provided the selectivity for the solvents analyzed, and cis-1,2-Dichloroethene Ethylbenzene
could be used for lists including lighter compounds, Dichloromethane p-Xylene
such as butane and propane. 1,4-Dioxane solution m-Xylene
Methanol o-Xylene
Residual Solvents Mixture - Class IIB PHR1065
Table 1. Accuracy and Reproducibility of the
Chloroform Nitromethane
SPME-GC/MS Analysis of Hemp Extract, Unspiked
1,2-Dimethoxyethane Pyridine
and Spiked at 10 µg/g
Hexane Tetralin 100
Class Amt. Measured in Avg. % 3-Methyl-2-pentanone Trichloroethylene
(per Unspiked Hemp Accuracy %
Solvent ICH) Extract (μg/g) (n=3) RSD Residual Solvents Mixture - Class IIC PHR1065
Methanol II 0 90% 3% N,N-Dimethylacetamide Formamide
Ethanol III 0 84% 9% N,N-Dimethylformamide 2-Methoxyethanol
Acetonitrile II 0 101% 6% 2-Ethoxyethanol N-Methylpyrrolidone
Acetone III 0 101% 5% Ethylene glycol Sulfolane
Isopropanol III 0 92% 8% SPME Fibers and Accessories
Tetrahydrofuran II 0 95% 7% SPME fiber assembly, Carboxen/Polydimethylsiloxane 57319
(CAR/PDMS), for use with autosampler
Hexane II 58.5 (13) NA —
SPME fiber assembly, Carboxen/Polydimethylsiloxane 57318
Cyclohexane II 0 101% 9% (CAR/PDMS), for use with manual sampler
Benzene I 0 105% 6% SPME fiber holder for CTC autosampler 57347-U
Heptane III 0 115% 7% SPME fiber holder for manual sampling 57330-U
Toluene II 0 103% 7% Capillary GC Column
m- & p-Xylenes* II 0 100% 6% Supel-Q™ PLOT, 30 m × 0.32 mm I.D., average 24242
o-Xylene II 0 119% 8% thickness 15 μm
*Measured together Accessories
NA: not analyzed due to high level detected in unspiked sample
Inlet Liner, Direct (SPME) Type, Straight Design, 0.75 2637501
Solvents in classes I, II and III by the International Conference on Harmonization
(ICH) guidelines.4 mm I.D. for Agilent GC
Molded Thermogreen® LB-2 Septa, with injection hole, 28336-U
11 mm (pk/50)
References
Headspace vial, screw top, rounded bottom, 10 mL, SU860099
1. Romano, L.L.; Hazekamp, A., Cannabis Oil: chemical evaluation of an clear glass (pk/100)
upcoming cannabis-based medicine. Cannabinoids, 2013, 1(1), 1–11.
Magnetic Screw Cap for Headspace Vials, PTFE/silicone SU860103
2. Sides, H., High Science. National Geographic, June 2015, 31–57. septum (pk/100)
3. Kemsley, J., Cannabis Safety. Chem. Eng. News, 2015, 93(16), 27–28.
4. International Conference on Harmonization (ICH) of Technical
Requirements for Registration of Pharmaceuticals for Human Use. Q3C Visit our cannabis testing resources at
(R5): Impurities: Guideline for Residual Solvents, Step 4, Feb 2011. SigmaAldrich.com/cannabis

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