www.lancasterlabs.com Title: Presentation title Document name :Powerpoint template.ppt EDR: Document owner: Author Last modified on: 28/05/2009
QUANTITATIVE ANALYSIS OF 15 SELECTED VOLATILE ORGANIC
COMPOUNDS IN MAINSTREAM CIGARETTE SMOKE
BY GCMS.
Norman Fraley
September 17, 2013
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The Objective
1) To extend our existing method to include additional FDA HPHC list compounds.
2) To apply this method to analyze smoke/vapor from: Burn-down cigarettes ISO/HCA Heat-not-burned cigarettes ISO/HCA e-cigarettes Modified HCA
3) Validate the method.
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The Challenge – New FDA Analytes
Vinyl chloride 2-Nitropropane 1,3 Butadiene Pyridine Ethylene oxide Ethylbenzene Isoprene Styrene Furan (not in smoke) Benzo[b]furan Propylene oxide Urethane Vinyl Acetate Nitrobenzene Acrylonitrile Quinoline Nitromethane Acetamide Benzene Acrylamide Toluene
Nine new volatile compounds added to the list
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Analyte Vapor Pressure (mm Hg at 25 ºC)
Vinyl chloride 2550 2-Nitropropane 18 1,3 Butadiene 1823 Pyridine 14 Ethylene oxide 1050 Ethylbenzene 9 Isoprene 550 Styrene 5 Furan* 493 Benzo[b]furan 0.4 Propylene oxide 433 Urethane 0.3 Vinyl Acetate 89 Nitrobenzene 0.2 Acrylonitrile 83 Quinoline 0.1 Nitromethane 82 Acetamide 0.04 Benzene 28 Acrylamide 0.01 Toluene 22
3 gases, 16 liquids and 2 solids at room temperature
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Analyte Vapor Pressure (mm Hg at 25 ºC)
Vinyl chloride 2550 2-Nitropropane 18 1,3 Butadiene 1823 Pyridine 14 Ethylene oxide 1050 Ethylbenzene 9 Isoprene 550 Styrene 5 Furan* 493 Benzo[b]furan 0.4 Propylene oxide 433 Urethane 0.3 Vinyl Acetate 89 Nitrobenzene 0.2 Acrylonitrile 83 Quinoline 0.1 Nitromethane 82 Acetamide 0.04 Benzene 28 Acrylamide 0.01 Toluene 22
3 gases, 16 liquids and 2 solids at room temperature
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Variables for Method Optimization
Smoking Machine: Linear or Rotary CFP size: 44mm or 92mm Trap Type: Impinger or XAD-4 or Carbon Solvent: MeOH EtoAc MTBE Impinger # 1 2 3 Trap temp -70oC 0oC Room Temp GC Column: Wax 624 DB other Extraction: shake stir sonicate GC Detectors: MSD NPD ECD GC Inlet Variables: Type Split Ratio Temperature
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Trap Types Examined
Cambridge Filter Pad
XAD-4 Sorbent
Carbon Sorbent
Tedlar Gas Trapping Bag
Impinger
Numerous historical methods use impingers, a proven methodology, how can we optimize?
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Example: Inlet Temperature
180
200
220
240
260
230/245
240
250
260
280
Tem
pera
ture
ºC
1.1500-1.2500
1.0500-1.1500
0.9500-1.0500
0.8500-0.9500
0.7500-0.8500
Relative Response
Relative Response Profile by Analyte 2013
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Method Synopsis
Smoke 5-20 cigs through CFP into MeOH Solution containing deuterated ISTD of EACH analyte.
Combine fluids with CFP, sonicate, chill, aliquot, inject.
Chilled <-70 C Room Temp
To Piston To Cigarette
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GCMS Method Conditions
15 deuterium labeled ISTD one for each analyte
Restek© VMS column
60m x 0.25mm x 1.4µm 36 cm/sec He constant flow SIM mode with qualifier ion
15.5 minute runtime
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Sample Chromatogram – 3R4F HCA
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Linearity – Instrument Sensitivity
1
10
100
1000
10000
100000
1000000
0.1 1 10 100 1000 10000 100000
Res
pons
e
Concentration (ng/mL)
Serial 10:1 Dilution
Vinyl Acetate
Acrylonitrile
Nitromethane
Benzene
Styrene
Benzofuran
The analytes span 4 orders of magnitude in our samples, so must our method.
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Reference Range
1
10
100
1000
10000
100000
Benzene
Pyridine
Ethylbenzene
Nitrobenzene
Quinoline
ng
3R4F HCA
1R5F ISO
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Validation
• Prepared a blend of analytes in MeOH/ISTD such that the low/high
level spike resulted in + 25% and + 50% of the nominal value of each analyte in each matrix/regime combination.
• Triplicate sample analysis, over multiple days and analysts. • Monitored sample stability both on-instrument and in storage • Parameters examined:
Specificity, Precision, Linearity, LOD/LOQ, Intermediate precision, Accuracy (Spiked Matrix to establish Recovery), Stability (sample and standards)
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Method Performance
HNB (35/60/2) 15 puffs
E-cig (55/30/3)
1st 20 puffs
HNB (55-30-2) 18 Puffs
1R5F (35/60/2)
3R4F (55-30-2)
RECOVERY RANGE Low 103% 101% 83% 85% 109% High 119% 120% 124% 121% 120%
PRECISION RANGE (n=3) Low 1% 0% 3% 0% 1% High 10% 19% 20% 10% 15%
Stability 32 Hours on instrument except for acrylamide 19 hours
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Conclusion
• By focusing on the lower vapour pressure volatiles we avoid sample preparation challenges that come from handling gaseous volatiles.
• Added 7 of the new FDA analytes.
• We can quantitatively determine 15 volatile organic compounds across 4 matrices and 5 smoking regimens with good resolution, accuracy and precision for each analyte.
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Acknowledgements
For myriad examples of support, patience and assistance: • Rachel Cone, Darius Grissom and Bhasha Desai Eurofins Lancaster Laboratories, Inc., Winston Salem, NC USA And
RJRT
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