Static Headspace (HS) Dynamic Headspace Spray-and-Trap (ST) SPME Membrane Inlet
Cigarette Cartridges and Liquids by Headspace Solid Phase ...
Transcript of Cigarette Cartridges and Liquids by Headspace Solid Phase ...
LABSTAT INTERNATIONAL ULC. 262 Manitou Drive
Kitchener, Ontario, Canada N2C 1L3 Phone: (519) 748-5409 Fax: (519) 748-1654 Web: www.labstat.com
Peter JOZA , David LI, Andrew MASTERS, Bill RICKERT
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Determination of Pyrazine Flavourants in Electronic
Cigarette Cartridges and Liquids by Headspace
Solid Phase Microextraction - Gas Chromatography-Mass Spectrometry (HS-SPME-GCMS)
2014 CORESTA CONGRESS October 12-16, 2014 Québec City, Canada
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Objectives
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Apply the HS-SPME-GCMS used for profiling volatile and semi-volatile compounds in tobacco products to e-liquids and e-cigarette aerosol
Quantify four targeted compounds:
2,3,5-Trimethylpyrazine (TriMPZ), 2,3,5,6-Tetramethylpyrazine (TetraMPZ), 2-Acetylpyrrole (APL) 2-Acetylpyrazine (APZ)
Provide some measure of the changes in the delivery of flavour components (targeted and non-targeted) during the “life” of an e-cigarette
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Background
67th TSRC: A rapid profiling method which can be applied to the detection of flavorings in various tobacco products
HS-SPME-GCMS Profiles Flavoring ID
Cigarettes, Cigar/Cigarillos
RYO, Pipe, Chew, Snus,
Snuff, Shisha, Dissovable Wiley, NIST/EPA/NIH, FFNSC
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Can the same process be used for the analysis of e-liquids and e-cig aerosols?
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Considerations
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The impact of the e-liquid matrix on the quantification and detection limits of flavour components using HS-SPME?
The type(s) and amount of flavour components
• Are these reflective of tobacco products?
The transfer of flavour components of an e-liquid
• Are these transferred to the e-cigarette aerosol?
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Sample Preparation for HS-SPME-GCMS Analysis
Base Method vs. Emissions / e-Liquids
ISTD/FPCS:
d6-Benzene
d5-Acetophenone
d5-Benzophenone
Add 20 μL ISTD/FPCS
HS-SPME-GCMS
Add 2 mL 3.0 M KClaq
0.5 g of tobacco filler in 10 mL HS vial
Cap the vial and keep at room temp for 2 hrs
Spike FLs std solution
5
Collection Pad
(up to 250mg)
Add 10 μL ISTD
ISTD/FPCS:
d5-2-ethylpyrazine
d7-quinoline
Chromatographic
Differences
Standard
Preparation
Only
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Instrument Parameters
Instrument Parameters
SPME Fiber 65 µm PDMS/DVB
Sample Ads Temp/Time 50 C/20 min
Sample Des Temp/Time 260 C/5 min
Fiber Condition Temp/Time 260 C/26 min
GC Column DB-WAXETR (30 m x 0.25 mm x 0.25 µm)
GC Injector Splitless mode, inlet temp 260 C.
MS Mode Full Scan (35 - 400 m/z)
GC Run Time 50 min (program 40 to 260°C)
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Instrument Parameters
Compound RT (min) Quantifier Qualifier
d5-2-ethylpyrazine 15.6 112 113
2,3,5-Trimethylpyrazine
(TriMPZ) 17.5 122 42
2,3,5,6-Tetramethylpyrazine
(TetraMPZ) 19.4 136 54
d7-quinoline 30.1 136 108
2-Acetylpyrrole (APL) 23.4 122 80
2-Acetylpyrazine (APZ) 30.9 94 109
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Instrument Parameters
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Nicotine
Propylene Glycol
Nicotyrine
Fibre Bleed Residual Solvent
~ 4ng TriMPZ
and TetraMPZ
~ 20ng APL
and APZ
Calibration Standard
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Instrument Parameters
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d5-2-ethylpyrazine
TriMPZ TetraMPZ
d7-quinoline
APL APZ
Nicotine
Propylene Glycol
Nicotyrine
Fibre Bleed Residual Solvent
~ 4ng TriMPZ
and TetraMPZ
~ 20ng APL
and APZ
Calibration Standard
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Determination of Matrix Effect
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Impact of Propylene Glycol and Glycerin
0
200000
400000
600000
800000
1000000
1200000
0 20 40 60 80 100
Res
po
nse
(A
rea)
Propylene Glycol (%)
TriMPZ
0
1000000
2000000
3000000
4000000
5000000
6000000
7000000
0 20 40 60 80 100
Res
po
nse
(A
rea)
Propylene Glycol (%)
APL
0.05
0.06
0.07
0.08
0.09
0.1
0.11
0 20 40 60 80 100
Res
po
nse
Rat
io
Propylene Glycol (%)
TetraMPZ/d5-2-EthylPZ
Calibration Standards Matrix
50:50 (v/v) Propylene Glycol to Glycerin solution
8 mg/mL Nicotine 250mg matrix at each level
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r2=0.997
Quantification of Target Analytes
LOD Determination - Root Mean Square Error (RMSE) *
Generation of a 4 or 5 point calibration curve
Instrument response (area counts) vs. standard concentration
Standards of concentration within an order of magnitude of estimated LOD (e.g. TetraMPZ - 1.10ng to 10.6ng)
* Handbook of Residue Analytical Methods for Agrochemicals, p68-70, Editor-in-Chief Dr Philip W Lee, DuPont Crop Protection,
USA; Published by John Wiley & Sons Ltd in 2003. 11
RMSE
LOD
LOQ
Compound LOQ (ng)
TriMPZ 4.27
TetraMPZ 2.75
APL 107
APZ 26.1
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Quantification of Target Analytes
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Compound Range
(ng)
TriMPZ 1 - 750
TetraMPZ 1 - 850
APL 10 - 8000
APZ 10 - 8200
d5-2-ethylpyrazine - 194ng d7-quinoline - 112ng
(ISTD for APZ)
All r2 > 0.99
TIC Response (semi-quantitative)
TriMPZ / TetraMPZ: 1 to 4µg
APL and APZ: 10 to 40µg
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Analysis of E-liquids – Experimental
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18 different formulations tested
• 13 e-liquids (1 advertized as “tobacco flavour)
• 1 e-liquid originating from a tobacco extract • 4 e-cigarette cartomizers (fresh)
• 4 variants of same product line (1 mentholated) • Centrifuge cartomizer at 1500 rpm for 5 min.
• 1 e-cigarette cartomizer (after use – first 20 puffs)
Single replicate for each formulation
• 0.25g sample size
Opportunity to identify other flavour components
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Analysis of E-liquids - Chromatography
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Sample 1
Tobacco Flavour
Sample 2
No Description
Sample 5
Americano
Sample 1
TriMPZ
Saturated Signal
Sample 2
Requires a more dilute sample for analysis
Sample 5
APZ
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Analysis of E-liquids - Results
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Sample No Matrix InfoTriMPZ
(ng/g)
TetraMPZ
(ng/g)
APL
(ng/g)
APZ
(ng/g)
1 E-Liquid Tobacco Flavour > UCL 197 - NQ
2 E-Liquid - 5.33 > UCL > UCL
3 E-Liquid Forest Fruit Mix 61.4 NQ - -
4 E-Liquid 7 Leaves 2601 > UCL - -
5 E-Liquid Americano 290 - NQ 8.75
6 E-Liquid Strawberry 5.13 NQ - -
7 Cubana 544 NQ - 195
8 E-Liquid 6.02 4.08 8767 1166
9 E-Liquid 6.21 4.94 - 31166
10 E-Liquid 45.9 7.07 NQ 402
11 E-Liquid 5.06 4.04 - NQ
12 E-Liquid 4.37 4.44 - 57.3
13 Tobacco Extract E-Liquid 52.7 NQ 129 -
14 Cartomizer Liquid 1 > UCL 3.7 NQ -
15 Cartomizer Liquid 2 > UCL NQ 106 -
16 Cartomizer Liquid 3 > UCL - - -
17 Cartomizer Liquid 4M > UCL NQ 117 -
18 Cartomizer Liquid (Used) > UCL - - -
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Analysis of E-liquids - Results
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Sample No Matrix InfoTriMPZ
(ng/g)
TetraMPZ
(ng/g)
APL
(ng/g)
APZ
(ng/g)
1 E-Liquid Tobacco Flavour > 16 µg 197 - NQ
2 E-Liquid - 5.33 68795 * 47656 *
3 E-Liquid Forest Fruit Mix 61.4 NQ - -
4 E-Liquid 7 Leaves 2601 3638 * - -
5 E-Liquid Americano 290 - NQ 8.75
6 E-Liquid Strawberry 5.13 NQ - -
7 Cubana 544 NQ - 195
8 E-Liquid 6.02 4.08 8767 1166
9 E-Liquid 6.21 4.94 - 31166
10 E-Liquid 45.9 7.07 NQ 402
11 E-Liquid 5.06 4.04 - NQ
12 E-Liquid 4.37 4.44 - 57.3
13 Tobacco Extract E-Liquid 52.7 NQ 129 -
14 Cartomizer Liquid 1 4625 * 3.7 NQ -
15 Cartomizer Liquid 2 4089 * NQ 106 -
16 Cartomizer Liquid 3 5605 * - - -
17 Cartomizer Liquid 4M 10716 * NQ 117 -
18 Cartomizer Liquid (Used) 3842 * - - -
* Estimated using TIC area count
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E – Cig Aerosol – Experimental
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1 brand (cartomizer 1 from E-liquids) • 4 replicates • 5 cartridges per replicate • Collection in 25 puff increments (5 increments) • Same cartomizers used for all 5 increments (20 total) • Freshly charged battery every 50 puffs • 44mm pad collection (½ pad SPME, ½ pad liquid extract)
Puffing Regimen • 70mL puff volume • 3.5 second puff duration (square wave profile) • 30 second puff frequency
Identify changes in delivery of flavour components (targeted and non-targeted)
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E – Cig Aerosol – Results
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Puff Segment TPM
(mg/cart)
Cartridge Wt. Loss (mg/cart)
Propylene Glycol
(mg/cart)
Nicotine (mg/cart)
Glycerin (mg/cart)
1 – 25
69.6
± 2.0
(2.9%)
67.5
± 1.8
(2.7%)
4.50
± 0.15
(3.4%)
3.02
± 0.10
(3.3%)
51.6
± 1.3
(2.5%)
26 – 50
65.1
± 2.0
(3.1%)
63.3
± 2.2
(3.5%)
4.27
± 0.10
(2.3%)
2.84
± 0.07
(2.5%)
48.5
± 1.6
(3.4%)
51 – 75
61.7
± 1.0
(1.6%)
60.2
± 1.1
(1.8%)
4.13
± 0.18
(4.4%)
2.72
± 0.11
(4.1%)
46.1
± 1.4
(3.0%)
76 – 100
58.2
± 1.2
(2.1%)
56.8
± 1.2
(2.1%)
4.06
± 0.06
(1.5%)
2.65
± 0.03
(1.2%)
44.3
± 0.7
(1.6%)
101 - 125
54.9
± 1.9
(3.4%)
54.1
± 1.9
(3.4%)
3.70
± 0.14
(3.7%)
2.42
± 0.09
(3.8%)
40.5
± 1.8
(4.4%)
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E – Cig Aerosol – Results
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Consistent loss in delivery of major aerosol components
Delivery of 5th segment is 80% that of the 1st segment (average)
75.0
80.0
85.0
90.0
95.0
100.0
2 3 4 5
Per
cen
tage
(%
) o
f Fi
rst
Pu
ff S
egm
ent
Puff Segment
Changes in Aerosol Delivery
MS TPM Cartridge Wt. Loss Propylene Glycol Nicotine Glycerol
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E – Cig Aerosol – Chromatography
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E-liquid
Collection Segment 1
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E – Cig Aerosol – Chromatography
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Only 2,3,5-Trimethylpyrazine (TriMPZ) found in the emmission
E-liquid
Collection Segment 1
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HS-SPME Results – Targeted
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Puff Segment Nicotine
(mg/cart) TriMPZ * (ng/cart)
1 – 25
3.02
± 0.10
(3.3%)
472
± 19
(4.0%)
26 – 50
2.84
± 0.07
(2.5%)
435
± 14
(3.3%)
51 – 75
2.72
± 0.11
(4.1%)
427
± 16
(3.7%)
76 – 100
2.65
± 0.03
(1.2%)
412
± 18
(4.0%)
101 - 125
2.42
± 0.09
(3.8%)
386
± 7
(1.8%)
* TriMPZ results extrapolated using TIC response
Delivery of TriMPZ in the 5th segment is 82% that of the 1st segment
Results are consistent with major components of e-cigarette aerosol
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E – Cig Aerosol – Chromatography
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No observed changes in TIC ratios amongst non-targeted compounds
Collection Segment 5
Collection Segment 4
Collection Segment 3
Collection Segment 2
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HS-SPME Aerosol – Non-Targeted
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0.0%
20.0%
40.0%
60.0%
80.0%
100.0%
120.0%
ETHYL ACETATE
ETHANOL
2-METHOXY-3-METHYLPYRAZINE
2,3,5-TRIMETHYLPYRAZINE
PROPYLENE GLYCOL
OXOPHORONE
BETA-DEMASCONEISOMER
2-HYDROXY-3-METHYL-2-CYCLOPENTEN-1-ONE
NICOTYRINE
ETHYLVANILLIN
Segment 2 Segment 5
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Summary
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HS-SPME-GCMS can be used for the determination of flavour components for both e-liquids and e-cigarette aerosol
• Optimization/Compromises dependent on levels
Matrix matching (glycerin / propylene glycol) need be considered for trace level quantitation
Screening of e-liquid is useful for identification of targeted compounds
This method can be used to assess the transfer of flavour components of an e-liquid to the E-cig aerosol.
• Targeted and non-targeted compounds
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Acknowledgements
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Mr. MingZhong Cui
Mrs. Helena Coetzer
Thank You !
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