DETECTION AND QUANTIFICATION OF NICOTINE IN ELECTRONIC CIGARETTES BY HPLC

Electronic cigarettes are a new and widely popular, but fairly

untested and unregulated technology. This project aims to
put electronic cigarette liquid manufacturers to the test by
determining the amount of nicotine in their products. Using
reverse-phased high performance liquid chromatography,
standard solutions of nicotine at concentrations of 50 ppm,
100 ppm, 200 ppm, 400 ppm, and 600 ppm were run in order
to determine a standard calibration curve. By comparing
these standards to five different electronic cigarette liquids,
nicotine was detected and quantified.
ABSTRACT
INTRODUCTION
Materials
Preparation of Nicotine Standards
Standards were prepared using a serial dilution method, with 50:50 MeOH H2O acting as
the solvent. Astock solution of 600 ppm nicotine was made from a Sigma-Aldrich 1mg/
ml nicotine solution. From this stock, samples of 400 ppm, 200 ppm, 100 ppm, and 50
ppm were created. Solutions were made in eppendorf tubes and filtered through 0.45m
filters using syringes upon transfer to HPLC vials.
Preparation of Electronic Cigarette Liquids:
Electronic cigarette liquids of two different flavors and brands were purchased. From
Savor Vapor, menthol refill liquid bottles of 0mg, 6mg, and 12mg were purchased, and
from iFill Vape, Bahama Mama liquids of 0mg and 6mg were purchased. Approximately
1ml of each concentration and flavor was filtered through a 0.45m syringe filter and
into HPLC vials.
Preparation of Mobile Phase:
An isocratic method was used, initially with a 25:75 MeOH H2O mobile phase (with
0.05% formic acid to adjust pH). Later runs, however, were done using a 20:80 MeOH
H2O mobile phase which was filtered prior to running.
MATERIALS AND METHODS
Given these results, we can conclude that there is, in fact, more nicotine per bottle of refill
liquid than these companies are claiming. Every liquid tested showed more nicotine than
advertised, including those labeled as nicotine free. We can therefore further conclude that
our hypothesis was correct.
DATA
CONCLUSION
REFERENCES
Setting for HPLC Reverse-phase
Column Zorbax Eclipse XDB-C8 (4.6 x 150mm) 5-micron
Mobile Phase 20:80 MeOH H2O (with 0.05% formic acid to adjust pH)
Injection Volume 5 l
Retention time 2 minutes
Wavelength 254 nm
Flow Rate 0.7 ml/min
OBJECTIVE/HYPOTHESIS
Objective:
To determine amount of nicotine in electronic cigarette refill
liquids.
Hypothesis:
We hypothesize that the electronic cigarette refill liquids are
improperly labeled and contain more nicotine than advertised.
ACKNOWLEDGEMENTS
RESULTS/DISCUSSION
Wed like to thank Dr. Malhotra for her constant counsel and support, Dr. Tannaci for
providing us with the chemicals we needed, Karen Kearsley of Agilent Technologies for
generously donating a column, and Dr. Cauchon for his HPLC expertise. Without the
guidance of these great mentors and professionals, our project would not have been the
great learning experience it was.
y = 9.5448x - 9.7338
R! = 0.9981
-1000
0
1000
2000
3000
4000
5000
6000
7000
0 100 200 300 400 500 600 700
P
e
a
k
A
r
e
a

Concentration (ppm)
Nicotine Standard Calibration Curve
Concentration
(ppm)
Nicotine (g) per
5 l Injection
Nicotine (mg) per
15 ml Bottle
Menthol 0mg 44.54 0.2227 0.681
Menthol 6mg 1330.172 6.651 19.953
Menthol 12mg 3987.691 19.938 59.814
BM 0mg 90.96 .453 1.359
BM 6mg 1469.906 7.35 22.05
0
10
20
30
40
50
60
70
Bahama
Mama
0mg
Bahama
Mama
6mg
Menthol
12mg
Menthol
6mg
Menthol
0mg
N
ic
o
tin
e
(m
g
) p
e
r
B
o
ttle

Electronic Cigarette
Comparison of Nicotine Levels
Labeled Amount
Amount We Detected
y = 0.005x
0
0.5
1
1.5
2
2.5
3
3.5
0 100 200 300 400 500 600 700
M
a
s
s
(u
g
) p
e
r
I
n
je
c
tio
n

Concentration (ppm)
Nicotine Standard Injection Curve
FURTHER WORK