Nicotine Retention Following Use of E-Cigarettes

1. Nicotine Retention Following Use of
E-Cigarettes
Dr. Nicole Tschierske, Dr. Grant O’Connell
Tobacco Campus Wiesbaden, 8 February 2016
1

2. Please note that the views and arguments presented in this paper have been designed to
encourage and stimulate debate and do not necessarily reflect Fontem Ventures' position
About Fontem Ventures B.V.
Innovative consumer goods company, developing
leading e-cigarette brands such as blu and JAI, as
well as a lifestyle energy brand Reon
Headquartered in Amsterdam, our more than 200
employees are active in the USA, UK, France and
Italy and in research labs in Hamburg, Beijing and
Silicon Valley
Fontem Ventures believes that changing consumer
lifestyles are creating vast new opportunities for
companies who can meet evolving needs. Fontem
is dedicated to developing and growing a portfolio
of consumable products that meet consumer
demands, today and tomorrow.
2

3. Please note that the views and arguments presented in this paper have been designed to
encourage and stimulate debate and do not necessarily reflect Fontem Ventures' position
Nicotine Retention and Potential Impact on
Indoor Air Quality
Why measuring nicotine
retention?
• Confirm nicotine transfer
from the aerosol into the
body
• Implications for
bystanders exposed to
2nd hand vapour
3
aerosol
compounds[inhaled]
compounds[released]
nicotine
uptake
so-called 2nd
hand vaping
Retention [%] =
[inhaled] − [exhaled]
[inhaled]
× 100 %

4. Please note that the views and arguments presented in this paper have been designed to
encourage and stimulate debate and do not necessarily reflect Fontem Ventures' position
Proton Transfer Reaction Mass Spectrometry
(PTR-MS)
4
Time-of-flight PTR-MS with custom-made interface for
A: mainstream e-cig aerosol analysis
B: exhaled breath following mouth-hold of e-cig aerosol
C: exhaled breath following inhalation of e-cig aerosol
A
B, C

5. Please note that the views and arguments presented in this paper have been designed to
encourage and stimulate debate and do not necessarily reflect Fontem Ventures' position
Application of PTR-MS in e-cig aerosol research
5
0 100 200 300 400 500
0
20
40
60
Concentration(ppbv)
Ion yield (ncps)
Nicotine
y = 0.1076 x + 0.4748
R
2
= 0.9998
Calibration plots nicotine including the
corresponding calibration functions, which have
been derived from linear fits of the data points
0 50 100 150 200 250 300 350 400
5x10
0
6x10
0
7x10
0
8x10
0
c)
b)
Time (s)
Nicotine
a)
4x10
3
6x10
3
8x10
3
Concentration(ppbv)
1x10
4
2x10
4
3x10
4
Concentrations (ppbv) of nicotine measured a) in the
mainstream aerosol, b) exhaled after aerosol mouth-hold and c)
exhaled after aerosol inhalation. Note the different orders of
magnitude for the concentration axes
Breiev et al., in press

6. Please note that the views and arguments presented in this paper have been designed to
encourage and stimulate debate and do not necessarily reflect Fontem Ventures' position
Nicotine retention studies with PuritaneTM and JaiTM
6
• Puritane Cartomiser
• Ad libitum puffs1
• Jai Cartomiser and
Clearomiser
• Defined puffs
2
• Jai Cartomiser and
Clearomiser
• Ad libitum puffs
3

7. Please note that the views and arguments presented in this paper have been designed to
encourage and stimulate debate and do not necessarily reflect Fontem Ventures' position
• Puritane Cartomiser
• Ad libitum puffs1
• Jai Cartomiser and
Clearomiser
• Defined puffs
2
• Jai Cartomiser and
Clearomiser
• Ad libitum puffs
3
Nicotine retention studies with PuritaneTM and JaiTM
7

8. Please note that the views and arguments presented in this paper have been designed to
encourage and stimulate debate and do not necessarily reflect Fontem Ventures' position
1 - Nicotine retention following use of Puritane™ e-cig
Methodology
8
O’Connell G, Colard S, Breiev K, Sulzer P, Biel SS, et al. (2015) An Experimental Method to Determine the Concentration of Nicotine
in Exhaled Breath and its Retention Rate Following Use of an Electronic Cigarette. J Environ Anal Chem 2: 161
Method: time-of-flight PTR-MS
Subjects: 3 experienced e-cig users (male, age 27-34)
Device: 8 mg/g, 16 mg/g and 20 mg/g nicotine ‘original’ flavoured
Puritane™ rechargeable e-cigarettes (cartomiser ‘closed’
system; PG:VG ≈ 70:30)
Vaping: ad libitum,
A: mouth-hold,
B: inhalation (5 reps. each), puff topography recorded via
SPA-M device
Nicotine calibration: liquid calibration in combination with smoking machine
operated e-cig aerosol trapped on to Cambridge filter pads
and subsequent GC-FID analysis of extracted filter solution (5
replicates)

9. Please note that the views and arguments presented in this paper have been designed to
encourage and stimulate debate and do not necessarily reflect Fontem Ventures' position
1 - Nicotine retention following use of Puritane™ e-cig
Results
nicotine concentration
(ppbv)
nicotine
retention
rate
(%)inhaled exhaled
8 mg/g 6317 1514 77
16 mg/g 13703 1514 89
20 mg/g 20668 1786 92
9
Mean concentration of nicotine delivered and exhaled and
nicotine retained for “mouth-hold” and “inhalation” across
different users at different nicotine levels
O’Connell G, Colard S, Breiev K, Sulzer P, Biel SS, et al. (2015) An Experimental Method to Determine the Concentration of Nicotine
in Exhaled Breath and its Retention Rate Following Use of an Electronic Cigarette. J Environ Anal Chem 2: 161
nicotine concentration
(ppbv)
nicotine
retention
rate
(%)inhaled exhaled
8 mg/g 6671 1.76 >99
16 mg/g 11468 2.31 >99
20 mg/g 18485 1.76 >99
“mouth-hold” “inhalation”

10. Please note that the views and arguments presented in this paper have been designed to
encourage and stimulate debate and do not necessarily reflect Fontem Ventures' position
• Puritane Cartomiser
• Ad libitum puffs1
• Jai Cartomiser and
Clearomiser
• Defined puffs
2
• Jai Cartomiser and
Clearomiser
• Ad libitum puffs
3
Nicotine retention studies with PuritaneTM and JaiTM
10

11. Please note that the views and arguments presented in this paper have been designed to
encourage and stimulate debate and do not necessarily reflect Fontem Ventures' position
2 - Nicotine retention following use of JAI™ e-cig
Methodology
11
Method: time-of-flight PTR-MS
Subjects: 4 experienced e-cig users (male, age 23-35)
Device: JAI™ cartomiser “closed” system and JAI™ tank “open” system
filled with 10 mg/g nicotine ‘original’ flavoured liquid (PG:VG ≈
70:30)
Vaping: A: mouth-hold,
B: inhalation
(6 reps. each; aerosol delivered in-mouth from smoking
machine operated e-cig; 3s puff duration, 55mL puff volume)
Nicotine calibration: liquid calibration in combination with device calibration via
nicotine in mainstream aerosol from smoking machine
operated e-cig (3s puff duration, 55mL puff volume, 6
replicates)

12. Please note that the views and arguments presented in this paper have been designed to
encourage and stimulate debate and do not necessarily reflect Fontem Ventures' position
2 - Nicotine retention following use of JAI™ e-cig
Results Cartomiser
12
0
200
400
600
800
1000
1200
1400
1600
JAI™ cartomiser
mouth-hold condition
0.00
0.50
1.00
1.50
2.00
2.50
3.00
3.50
4.00
4.50
JAI™ cartomiser
inhalation condition
Nicotine
concentration (in
ppbv) in exhaled
breath per subject
(smoking machine
operated e-cigs)
Error: standard deviation
of n= 6 replicates
concentration of
nicotine delivered in-
mouth of subjects
nicotine retention 98% 75% 94% 92% 100% 100% 100% 100%
4999±251 ppbv 6765±519 ppbv 4999±251 ppbv 6765±519 ppbv

13. Please note that the views and arguments presented in this paper have been designed to
encourage and stimulate debate and do not necessarily reflect Fontem Ventures' position
2 - Nicotine retention following use of JAI™ e-cig
Results Clearomiser
13
Nicotine
concentration (in
ppbv) in exhaled
breath per subject
(smoking machine
operated e-cigs)
Error: standard deviation
of n= 6 replicates
concentration of
nicotine delivered in-
mouth of subjects
nicotine retention 78% 73% 84% 71% 100% 100% 100% 100%
12172±1344 ppbv 10146±397 ppbv 12172±1344 ppbv 10146±397 ppbv
0
1000
2000
3000
4000
5000
JAI™ clearomiser
mouth-hold condition
0.00
1.00
2.00
3.00
4.00
5.00
6.00
JAI™ clearomiser
inhalation condition

14. Please note that the views and arguments presented in this paper have been designed to
encourage and stimulate debate and do not necessarily reflect Fontem Ventures' position
• Puritane Cartomiser
• Ad libitum puffs1
• Jai Cartomiser and
Clearomiser
• Defined puffs
2
• Jai Cartomiser and
Clearomiser
• Ad libitum puffs
3
Nicotine retention studies with PuritaneTM and JaiTM
14

15. Please note that the views and arguments presented in this paper have been designed to
encourage and stimulate debate and do not necessarily reflect Fontem Ventures' position
3 - Nicotine retention following use of JAI™ e-cig
Methodology
15
y = 786.05x - 620.75
R² = 0.999
1500
2000
2500
3000
3500
2.5 3.5 4.5 5.5
nicotineconcentration
inppbv
puff duration in s
Method: time-of-flight PTR-MS
Subjects: 4 experienced e-cig users (male, age 23-35)
Device: JAI™ cartomiser “closed” system and JAI™ tank “open” system
filled with 10 mg/g nicotine ‘original’ flavoured liquid (PG:VG ≈
70:30)
Vaping: ad libitum with vaping topography being recorded by a SPA-M
device
Calculation of inhaled
nicotine concentration
via device calibration
using puff duration
recorded with SPA-M device

16. Please note that the views and arguments presented in this paper have been designed to
encourage and stimulate debate and do not necessarily reflect Fontem Ventures' position
3 - Nicotine retention following use of JAI™ e-cig
Results Cartomiser
vaping condition
nicotine concentration
inhaled in ppbv
nicotine concentration
exhaled in ppbv
nicotine
retention in %
Subject 1
mouth-hold 8158 (± 1952) 996 (± 299) 87,8 (± 2,26)
inhalation 6108 (± 892) 5,01 (± 0,60) 99,9 (± 0,02)
Subject 2
mouth-hold 4630 (± 573) 312 (± 116) 93,3 (± 2,13)
inhalation 3898 (± 2358) 6,63 (± 0,99) 99,7 (± 0,29)
Subject 3
mouth-hold 1637 (± 265) 753 (± 236) 53,1 (±15,8)
inhalation 1407 (± 380) 5,06 (± 1,06) 99,6 (± 0,12)
Subject 4
mouth-hold 1952 (± 209) 843 (± 106) 56,7 (± 5,16)
inhalation 1906 (± 236) 6,98 (± 1,85) 99,6 (± 0,13)
16
Mean concentration of nicotine delivered and exhaled and nicotine retained for “mouth-hold” and
“inhalation” for different users; numbers in brackets represent standard deviation of 6 replicates

17. Please note that the views and arguments presented in this paper have been designed to
encourage and stimulate debate and do not necessarily reflect Fontem Ventures' position
3 - Nicotine retention following use of JAI™ e-cig
Results Clearomiser
17
Mean concentration of nicotine delivered and exhaled and nicotine retained for “mouth-hold” and
“inhalation” for different users; numbers in brackets represent standard deviation of 6 replicates
vaping condition
nicotine concentration
inhaled in ppbv
nicotine concentration
exhaled in ppbv
nicotine
retention in %
Subject 1
mouth-hold 4547 (± 716) 218 (± 49,5) 95,2 (± 0,89)
inhalation 5711 (± 1503) 4,26 (± 1,12) 99,9 (± 0,04)
Subject 2
mouth-hold 6794 (± 568) 601 (± 222) 91,2 (± 3,21)
inhalation 4876 (± 901) 4,94 (± 0,32) 99,9 (± 0,02)
Subject 3
mouth-hold 3276 (±168) 177 (± 24) 94,6 (± 0,45)
inhalation 2456 (± 644) 4,90 (± 0,54) 99,8 (± 0,04)
Subject 4
mouth-hold 3770 (± 349) 639 (±63,3) 83,0 (± 1,96)
inhalation 3594 (± 189) 0,84 (± 0,09) 100 (± 0,00)

18. Please note that the views and arguments presented in this paper have been designed to
encourage and stimulate debate and do not necessarily reflect Fontem Ventures' position
Summary
- Time-of-flight PTR-MS allows exact quantification of inhaled and exhaled nicotine
concentration following use of e-cigarettes.
- More than 99% of nicotine is retained when aerosol is inhaled; if held in-mouth only,
retention is highly subject dependent and varies between 50-94%.
 The negligible quantities of nicotine exhaled have no impact on indoor air quality
when compared to regulatory standards (see: O’Connell et al. 2015 IJERPH)
 This is in alignment with Public Health England report 2015: “EC use releases
negligible levels of nicotine into ambient air with no identified health risks to
bystanders”.
- Future work: retention of base liquid components (PG, VG), aroma compounds
18

19. Please note that the views and arguments presented in this paper have been designed to
encourage and stimulate debate and do not necessarily reflect Fontem Ventures' position
Acknowledgement
19
Kos Breiev
Dr. Grant O‘Connell
Dr. Philipp Sulzer
Dr. Kerstin Burseg