1. POLYCYCLIC AROMATIC
HYDROCARBONS IN THE
MAINSTREAM SMOKE OF
POPULAR U.S.
CIGARETTES
BY AN T. VU, KENNETH M. TAYLOR, MATTHEW R.
HOLMAN, YAN S. DING, BRYAN HEARN, CLIFFORD
WATSON
ACS JOURNAL OF CHEMICAL RESEARCH IN
TOXICOLOGY
JULY 30, 2015
PRESENTED BY: ROWENA FERRER
1

2. TOBACCO SMOKE
 Leading cause of PREVENTABLE death
 Six million annual mortalities
 One half trillion dollars of economic damage worldwide each
year
 Complex composition of cigarette
smoke includes many toxic
substances
2

3. OUTLINE
 What are Polycyclic Aromatic Hydrocarbons?
 Sample Preparation
 Smoke Collection
 Solid Phase Extraction
 Gas Chromatography/Mass Spectrometry
 Results
 Conclusion
3

4. POLYCYCLIC AROMATIC
HYDROCARBONS
Benzo[a]pyren
e
(BaP)
IARC
4

5. POLYCYCLIC AROMATIC
HYDROCARBONS
5
 Environmental pollutants
 Found in tar, crude oil
 PAHs do not naturally occur
in tobacco plant
 Complete Combustion:
 Hydrocarbon + O2  CO2 +
H2O
 Incomplete Combustion:
 Hydrocarbon + O2  CO +
C + H2O

6. POLYCYCLIC AROMATIC
HYDROCARBONS
 Also, found in some
unburned tobacco
products
 Pipe tobacco
 Chewing tobacco
 Snuff
 fire cured tobacco
varieties: slow oxidation to
reduce chlorophyll content
for flavor
6

7. OVERVIEW
7
ISO CI
PUFF INTERVAL 60 s 30 s
PUFF DURATION 2 s 2s
VOLUME 35 mL 55 mL
VENTILATION UNBLOCKED BLOCKED
FDA requires HPHC tobacco testing and reporting
1. Levels of 14 PAHs in the smoke of 50 commercial
U.S. cigarette products using GCMS
 International Organization of Standardization (ISO)
 Canadian Intense (CI)
2. Potential correlations between
BaP and other individual PAHs

8. MATERIALS
 35/50 cigarettes used, make up 54% of the
total U.S. cigarette market share based on
sales volume
 The 50 cigarettes consisted of 23 brands
with various sub-brands, lengths, sizes,
and package types
 Regular, menthol, or Turkish flavored
 All contained a cellulose acetate filter
 3R4F Reference cigarettes from University
of Kentucky
8

9. R² = 1
0
5
10
15
20
25
30
35
0 2 4 6 8
Signal
Concentra on
Calibra on Curve
Peak Area
Peak Area
METHODS
 Calibration Curve
 Varying amounts
of standard stock
solution containing
all 14 PAHs was
prepared in
methanol
 25 μL of 13C PAH
internal standard
solution
9

10. 3R4F REFERENCE
CIGARETTE10
 Research standard
designed to represent
tobacco composition of US
consumer tobacco
products
 Do not include any
additional flavorings
 Smoked as QC samples
during each smoking run

11. SAMPLE PREPARATION
11
 Cigarettes smoked to 23 mm to butt length in
ASM500 16-port smoking machine
 ISO
 CI
 Each CFP was placed in 10 mL of methanol in 13
mL vial and then decanted.
 Zymark LV Evaporator to reduce volume to 2 mL.
 Any remaining particulate matter was removed
with 0.45 μL nylon syringe filter.
 3.5 mL HPLC grade water

12. 12
SOLID PHASE EXTRACTION
GILSON 215
SPE

13. 13
SELECTED ION MONITORING
GC/MS
The cyclohexane extracts were
evaporated to approximately 0.5 mL, and
a 1 μL aliquot was used for GC/MS
analysis
AGILENT
6890/5973

14. QUADRUPOLE MASS
ANALYZER14
SELECTED
ION
MONITORIN
G
Current

15. ADVANTAGES & DISADVANTAGES
OF GC/MS
 Advantage
 Good at separating
complex mixtures into
components
 Disadvantage
 Possibility of co-elution
 Advantage
 Most powerful detector
for chromatography
 Disadvantage
 Possibility of similar
patterns of ionized
fragments
15
GAS CHROMATOGRAPHY
MASS SPECTROMETRY

16. PAH LEVELS IN 3R4F AND 2R4F REFERENCE
CIGARETTES
16
*Y axis depicts PAH yield on a log-2 scale
Determination of 14 PAHs in Mainstream Smoke from Domestic Cigarettes,

17. Coefficients of Variance
17

18. TOTAL PAH YIELDS
18
Abbreviated: American Spirit Blue

19. AMERICAN SPIRIT BLUE
19
 Tobacco Mass 881 mg/cig
 216 mg more than average
tobacco mass of other 49 cig (665
mg/cig)
 Flue Cured Tobacco
 Produces higher molecular

20. AVERAGE YIELDS OF INDIVIDUAL PAHs
FOR ALL 50 COMMERICAL CIGARETTES
20
log2scale

21. SPIDER CHART SHOWING BaP
COORELATION WITH INDIVIDUAL AND
TOTAL PAHs21

22. LINEAR RELATIONSHIPS BETWEEN BaP
YIELDS WITH OTHER PAHs IN ISO
REGIMEN22

23. LINEAR RELATIONSHIPS BETWEEN BaP
YIELDS & OTHER HIGH MOLECULAR WT. PAHs
USING CI REGIMEN23

24. TO CONCLUDE…
24
 Survey of PAH mainstream smoke yields for
popular U.S. cigarettes having a wide range of
smoke deliveries
 All 14 PAHs found in all 50 test cigarettes
 American Spirit Blue, Winston Red, Winston Red
100s
 Carlton White 100s & Marlboro Silver
 PAH yield vs molecular weight
 Very strong correlations between BaP and those
with similar molecular weight and chemical
structures

25. QUESTIONS…
25

26. Cigarette Model
26

27. STATISTICS
 P-Value: measure of statistical significance of
correlation (>0.5)
 Coefficients of determination (r2): measure of
the goodness of fit
 Between smoke deliveries of BaP and and other
PAHs on on the regression lines
27

28. PEARSON CORRELATION
COEFFICIANT (r)28

29. 29
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