LABSTAT INTERNATIONAL ULC.262 Manitou Drive

Kitchener, Ontario, Canada N2C 1L3Phone: (519) 748-5409 Fax: (519) 748-1654 Web: www.labstat.com

Xinyu LIU, Peter JOZA, Bill RICKERT

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Determination of Allyl Alcohol in Electronic Cigarette (E-cig) Aerosol and Liquid Using Gas

Chromatography-Mass Spectrometry Method

69th TOBACCO SCIENCE RESEARCH CONFERENCESeptember 20-23, 2015

Naples, Florida USA

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Objectives

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➢ To develop a method for the analysis of allyl alcohol in various matrices.

➢ To demonstrate that smoke methodologies are not directly applicable to e-cigarette aerosols.

➢ To determine the amount of allyl alcohol in cigarette smoke, e-cigarette aerosols and liquid.

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Ally alcohol in e-liquid or

aerosol

Residue of allyl alcohol used for synthesizing glycerol

Glycerol breaks down into allyl alcohol catalyzed by trace metals

or acids

Allyl Alcohol Possible Sources

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Approaches for Allyl Alcohol Analysis

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Authors & Journal Target Samples Method

EPA method 1624 Environmental samples Purge &Trap-GC/MS

EPA method 8240 Environmental samples Purge &Trap or direct injection-GC/MS

OSHA PV2140 Environmental samples TD-GC-FID

NIOSH Method 1402 Environmental samples TD-GC-FID

Stuart et al.Coresta 2013 E-cigarette aerosol screen TD-GC-TOFMS

Chris et al.TSRC 2014 E-cigarette emission screenTD-GC-TOFMS/FID

+ Heart Cutting

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Experimental Steps

Accurately weigh 0.1g solution

Filter through 0.22 µm syringe filter

Add 5 mL Methanol

Add methanol to 10 mL mark

Vortex and hand-shake 1 minute

Filtrate for GC-MS analysis

Add 0.1 mL ISTD

E-liquid Solution Samples

Transfer Pad into Erlenmeyer flask

Filtrate for GC-MS analysis

Transfer impinger solution into flask

Extract via Shaker 30 minutes

Filter through 0.22 µm syringe filter

Add accurately 0.2 mL ISTD

Aerosol Samples

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Analytical Conditions

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Instrument ParametersColumn ZB-WAX 60 m X 0.25 mm X 0.25 µm

Injector temperature 250°C

Oven temperature 70 °C for 4 minutes; 10 °C per minute to 220 °C; hold 3 minutes; 25 °C per minute to 250 °C; hold 2 minutes

Column flow 1mL/min

Injection method Split mode with split ratio 10:1

Injection volume 1 µL

Transfer line temperature 250°C

Source temperature 230°C

MS quad 150°C

Acquisition mode SIM

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MS Quantitation Parameters

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Component RT Quantifier ion Dwell time

Qualifierion

(min) (m/z) (ms) (m/z)

Allyl Alcohol 7.315 57 100 58

D5-Allyl Alcohol (ISTD) 7.219 61 100 62

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Allyl Alcohol Identification-Full Scan

Column: HP-PLOT/U 30mX0.320mmX10.0umOven program: 45°C hold 1.5 min, 3°C/min to 105°C; 25°C/min to 135°C hold 5 min; 30°C/min to 190°C hold 15 min.

3R4F Cig Smoke

Match factor: 772R. Match factor : 869Prob.(%): 69.1%

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Sample Type Description Response Area (m/z=58)

Response Area (m/z=57)

Response Ratio (57/58)

10% Range*

Standard Solution Std1 (1869ng/ml) 17694 100805 5.70

Std2 (748 ng/ml) 6722 38695 5.76

Std3 (374 ng/ml) 3147 19011 6.04

Std4 (187 ng/ml) 1655 9939 6.01

Std5 (74.8ng/ml) 579 3503 6.06

Std6 (29.9ng/ml) 177 1170 6.61

Average 6.03 5.43~6.63

Allyl Alcohol Identification-SIM

*Guidance document on analytical quality control and validation procedures for pesticide residues analysis in food and feed. SANCO/12495/2011 Implemented by 01/01/2014

E-cig aerosol Replicate-1 544 3401 6.25

Replicate-2 797 5051 6.34

Average 6.24

3R4F smoke Replicate-1 7540 47099 6.25

Replicate-2 7713 47970 6.22

Replicate-3 8386 51003 6.08

Average 6.23

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Distribution of Allyl Alcohol Between Pad and Impinger Solutions

E-cigarette

CF Pad (A)

Impinger Containing Methanol(B)

To Second Impinger

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Distribution of Allyl Alcohol Between Pad and Impingers Solution

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CambridgeFilter Pad Impingers Pad+Impingers

(Total)Pad/

(Pad+Impingers)[ng] [ng] [ng] [%]

E-cig Aerosol (example)

Observation1 15.2 647 662 2.29

Observation 2 47.6 723 771 6.18

Observation 3 70.8 689 760 9.32Observation 4 170 444 614 27.7

Average 702 (CV10.8%) 11.4 (CV 98.9%)

3R4F SmokeObservation1 7815 25770 33585 23.3

Observation2 8923 29040 37963 23.5Observation3 9149 25870 35019 26.1

Average 35522 (CV 6.3%) 24.3 (CV 6.4%)

* 55/3/30 “ square wave” profile puffing regimen (volume/duration/frequency) with a 100 puff collection

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Composition Comparison for E-liquid, E-cig Aerosol and Cigarette Smoke

Data source: R. Tayyarah, G.A. Long, Regulatory Toxicology and Pharmacology, 70(2014)704-710

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Changes in E-cig Aerosol CollectionGuidelines

➢The composition of the delivered E-cig aerosol is significantly different from conventional cigarette smoke and might affect the distributions of target analytes between pad and impingers.

➢Developing new sample trapping mode (e.g. without pad) or device for e-cig studies might be necessary to simplify sample collection procedures.

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LOD/LOQ Various Determination Guidelines➢Signal to Noise

➢Linear Regression

➢Standard Deviations of Lowest Calibration Concentration

➢Blank Determination

➢Precision-based Approaches

➢Limit of Blank• • • • • • •

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Allyl Alcohol Estimated LOD/LOQ Comparisons

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Method Replicates LOD LOQ

[n] [ng/cig] [ng/cig]Signal-to-noise

10 3.62 12.1

Linear regression

3 18.2 60.6

Lowest Concentration

10 3.14 10.5

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Method Characteristics

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E-cig Liquid Solution

Recovery (Accuracy)

Laboratory Reagent Blank (LRB) Not Determined

Laboratory Fortified Blank (LFB) (%) 102±3.3

Laboratory Fortified Matrix (LFM) (%) 103±4.4

Detection Limit

Limited of Detection (LOD) 314 ng/g

Limited of Quantification (LOQ) 1050 ng/g

E-cig Aerosol

Not Determined

101±3.6

107±3.6

62.8 ng/collection

210 ng/collection

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4.0 5.0 6.0 7.0 8.0 9.0 10.0 11.0 12.0 13.0 Min

4.0 5.0 6.0 7.0 8.0 9.0 10.0 11.0 12.0 13.0 Min

Abundance

Abundance

2000

1000

2000

1000

Allyl Alcohol

D5-Allyl Alcohol

E-cigarette Solution Analysis

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2000

Abundance

Abundance

3.0 4.0 5.0 6.0 7.0 8.0 9.0 10.0 11.0 12.0 13.0 14.0 15.0 Min

3.0 4.0 5.0 6.0 7.0 8.0 9.0 10.0 11.0 12.0 13.0 14.0 15.0 Min

1000

1000

2000

Allyl Alcohol

D5-Allyl Alcohol

E-cigarette Aerosol Analysis

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Mainstream Smoke Analysis (3R4F)

3.0 4.0 5.0 6.0 7.0 8.0 9.0 10.0 11.0 Min

3.0 4.0 5.0 6.0 7.0 8.0 9.0 10.0 11.0 Min

Abundance

Abundance

3000

1000

1000

3000

Allyl Alcohol

D5-Allyl Alcohol

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Allyl Alcohol in Aerosol, Liquid & Smoke

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SampleType

Sample Collection

Regime

Replicates[n] Mean Std. Dev. RSD

[%]

E-cig liquid NA 5 3733 ng/g 497 ng/g 13.3

* 80/3/30 “ square wave” profile puffing regimen (volume/duration/frequency) with a 100 puff collection

E-cig aerosol 80/3/30* 5 1510 ng/collection 163 ng/collection 10.8

3R4F smoke HCI 5 12018 ng/cig 574 ng/cig 4.8

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Summary

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➢The developed method can be applied to identify and quantitatively analyze allyl alcohol in e-cigarette aerosol, e-liquid solution and cigarette smoke.

➢The results indicated that the estimated LOD/LOQ were not always equivalent. It is therefore essential to have a clearly described procedure for estimating the LOD/LOQ during method validation to allow interlaboratory comparisons.

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Acknowledgements

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➢ Labstat International ULC Scientists, Analysts and Technicians

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