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Analytical Chemistry Letters
ISSN: 2229-7928 (Print) 2230-7532 (Online) Journal homepage: http://www.tandfonline.com/loi/tacl20
Nicotine Estimations in Electronic Cigarette E-
Liquids Among Malaysian Marketed Samples
Azizur Rahman, Mohamad Haniki Nik Mohamed & Syed Mahmood
To cite this article: Azizur Rahman, Mohamad Haniki Nik Mohamed & Syed Mahmood (2018)
Nicotine Estimations in Electronic Cigarette E-Liquids Among Malaysian Marketed Samples,
Analytical Chemistry Letters, 8:1, 54-62
To link to this article: https://doi.org/10.1080/22297928.2017.1400920
Published online: 15 Mar 2018.
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Nicotine Estimations in Electronic Cigarette
E-Liquids Among Malaysian Marketed Samples
Azizur Rahman 1*, Mohamad Haniki Nik Mohamed 1, Syed Mahmood 2*
1 International Islamic University Malaysia - Pharmacy, Sultan Ahmed
Shah bandar Indera Mahkota Kuantan Pahang 25200, Malaysia
2 University Malaysia Pahang - Department of Pharmaceutical Engineering,
Faculty of Engineering and Technology, Gambang, Malaysia
Abstract: E-cigarette (e-cig) is becoming a popular substitute for tobacco cigarette all over the world.
The chief constituents of e-cig e-liquids are nicotine, propylene glycol, glycerine, and a couple of other flavouring
constituents. The current study objective was to find out the discrepancy in the labelled vs actual nicotine
concentration amid the Malaysian marketed e-liquids brands for consumer safety purpose. The study examined
the 69 e-liquids samples that were selected from Malaysia market. The nicotine estimation was measured by
Agilent 6890N Gas chromatography-mass spectrometry with 5973 mass selective detector (MSD). Out of 69
samples analysed for actual nicotine level, in 64 samples nicotine concentration was found to be as low from -
1 to -67.13 % and in remaining 5 samples it contained higher from +2.8 to +24.70. A total of 60 (86.9 %) samples
were found to have significant discrepancies in labelled vs. actual nicotine concentration. The study concludes
that high discrepancy in the labelled vs actual nicotine concentration was found among the Malaysian marketed
e-liquids brands. Based on analysis results, there is an urgent need for regulation for these marketed nicotine e-
liquids products in Malaysia. The study suggests that corresponding Malaysian tobacco regulatory authorities
should instruct suppliers of e-liquids for flawless packaging for consumer safety reason.
Keywords: Electronic cigarette, tobacco cigarette, e-liquids, nicotine, concentration.
Introduction
E-cigarette (e-cig) is becoming a popular sub-
stitute for tobacco cigarette all over the world and
Malaysia is not an exception to this 1-3. E-cig is
comparatively novel and its consumption is
spreading as electronic nicotine delivery system
(ENDS) worldwide. It is a battery-controlled in-
strument available in the various sizes and shapes
like conventional cigarettes, pens, and boxes 4.
Herbert A Gilbert primarily invented the notion of
e-cig in 1965. However, the current design of e-
cig endorsed to Mr Hon Lik, a Chinese chemist 5.
The gadget first announced in the Chinese mar-
ketplace in May 2004 as an aid for the tobacco
cigarette (TCG) abstinence then carries across
to the different nations. The aim to developed e-
cig was to simulate the act of smoking by way of
nicotine supply but without the noxious effects of
tobacco smoke which are supposed to be practi-
cally accountable for all TCG destructive effects
6, 7. The vapours, which produce from the e-cig,
resembles smokes that exchanges most of the
social and behavioural features connected with
smoking. The e-cig does not release smoke but
vaporises the e-liquids solution that vapers inhale.
The chief constituents of e-cig liquids are nico-
ISSN Print: 2229-7928
ISSN Online: 2230-7532
Received 02 June 2017; accepted in revised form 31 October 2017
*Corresponding authors (Azizur Rahman and Syed Mahmood )
E-mail: < aziznoor51@gmail.com; syedmahmood@ump.edu.my > © 2018, Har Krishan Bhalla & Sons
TACL 8 (1) 2018 pp 54 - 62 54
tine, propylene glycol, glycerine, and a couple of
other flavouring constituents. The quantity of nico-
tine in the e-liquids solutions is generally offered
from 6 to 24 mg in the marketed available e-liq-
uids samples 7, 8. However, certain preceding stud-
ies exposed that the marketed e-liquids samples
found to have variations in the quality along with
discrepancies in the labelled vs actual nicotine
concentration 8-10. Therefore, there is no assur-
ance that the consumer’s daily used e-liquids
samples contain accurate nicotine concentration
with the unadulterated grade. Consequently, be-
cause of the wrong nicotine level in e-liquids con-
sumers assume that the e-cig is less pleasing and
unproductive substitute for the tobacco cigarette.
The reasons for inconsistency of nicotine con-
centration in e-liquids may alarm adsorption of
nicotine with packaging material or it could be in-
teraction with other ingredients of e-liquids or else
it may imply to improper handling and storage.
The regulation of e-cig in many countries is still
unsettled due to the paucity of sufficient knowl-
edge about the effectiveness and safety of e-liq-
uids 6,8. This is chiefly concerned when the ma-
jority of the e-liquids manufacturers do not reveal
the information relating to their products manu-
facturing process.
Presently, in Malaysia, under the poisons act
1952 and control of the drugs and cosmetics regu-
lations, nicotine is classified as a class C poison.
Devices without nicotine are classified as electri-
cal appliances. The sale, distribution and importa-
tion of unlicensed nicotine containing e-cigs are
prohibited. The act also forbids the sale and sup-
ply of this poison to adolescents under the age of
18 years old. Nicotine-containing e-cig can only
be sold by the licensed pharmacies by the pre-
scription of registered medical practitioners 11, 12.
The unexpected popularity of e-cig emerging as
a challenge to the current tobacco regulatory
framework, which design based on the conven-
tional tobacco cigarette. The entry of e-cig in the
open market may suggest the re-evaluation of the
nicotine-containing products for consumer’s well-
being.
Presently, in Malaysia, the chief apprehension
related to this accessible nicotine-containing prod-
uct is the lack of manufacturing guidelines for
national and as well as for the international avail-
able e-liquids brands 13. In Malaysia, the majority
of e-liquids samples which currently available are
mainly imported from China. Yet many Malay-
sian made refilled e-liquids samples are too avail-
able in the market. So far, no major studies have
carried out to assess the actual nicotine concen-
tration amongst Malaysian marketed e-liquids
brands. Therefore, the current study objective was
to find out the discrepancy in the labelled vs ac-
tual nicotine concentration in Malaysian marketed
e-liquids samples for customer’s safety purpose.
Materials and methods
Sample selection
The study examined the 69 e-liquids samples
that were selected from the Malaysia market. The
selection of the e-liquids for analysis was based
on discussion and suggestion from the prominent
e-liquid suppliers on regional levels. After receiv-
ing e-liquids from the suppliers, each sample was
given a unique code number and stored in original
containers. All samples were refrigerated between
4-8°C prior to analysis to restrain nicotine degra-
dation.
Preparation of standard nicotine and inter-
nal standard quinoline
Control and seven nicotine standard solutions
(Acros Organics, 99 %) were prepared in ranged
from 25 μg to 2 mg/ml. The base of standard and
control solutions was composed of 49 % glycer-
ine, 49 % Propylene Glycol and 2 % water. The
Internal Standard (IS) quinoline (across organics,
99 %) was prepared in the concentration of 50
mg/ml by adding 457 μl to 10 ml methanol. IS of
10 μl was added to all nicotine standard solu-
tions and run in triplicate. To ensure accurate
results for all samples, the calibration curve had
the linear coefficients of0 R2 > 0.99 or above
(Figure 1).
Sample preparation
All samples were prepared as per the method
described in Goniewicz et al. 8 with a small modi-
fication. Samples solutions were prepared by add-
ing directly 15μl of e-liquid into GC amber coloured
vial then added the 10 μl internal standard (IS)
and finally made up the volume upto 1000 μl by
HPLC grade methanol. All vials were sonicated
Azizur Rahman et al., / TACL 8 (1) 2018 54 - 62 55
for 20 minutes and vortexed for 10 seconds at
3000 rpm to ensure proper mixing of all added
ingredients. All samples were analysed in tripli-
cate. All e-liquid samples were diluted with metha-
nol in 1:100 ratios. This dilution was carried out
due to the below mention reasons.
The majority of the commercially purchased e-
liquids were very viscous in nature, so without
dilution, this viscous solution can form air bubbles
in GC-MS autosampler syringe and may induce
choking. A 1:100 dilution eliminates any viscosity
issues. The selected refilled e-liquids had nicotine
concentration of 3-18 mg/ml, which was outside
the level of the calibration curve (25 mcg - 2 mg).
Therefore, a 1:100 dilution resulted in nicotine con-
centration within calibration range.
Nicotine estimation by GC-MS
The nicotine estimation was measured by
Agilent 6890N Gas chromatography with 5973
mass selective detector (MSD). A capillary col-
umn of 30 m × 0.32 × 0.25 μm (Agilent) with
helium flow rate of 2.4/min was used. Injector
and detector temperature was kept 300°C and
column temperature increased from 60 to 200°C
(20°C/min) and finally held constant for 5 min-
utes. The sample injection volume was 1 μl. The
retention times for quinoline and nicotine were
4.234 and 4.944 respectively.
Statistical analysis
A mean average of nicotine concentration in
each sample, standard deviation and their relative
variances were calculated. The percentage diffe-
rences for every sample were calculated by com-
paring the labelled amount of nicotine with the
actual estimated amount. The p-value determined
by the one sample t-test which < 0.05 were con-
sidered significant. The analysis was performed
by using statistical package for social sciences
(IBM®, SPPS® Inc., Chicago, IL) version 21.
Results
The actual nicotine concentration in 69 samples
of the selected refilled e-liquids from Malaysian
market is presented in Table 1. Out of 69 samples
analysed for the actual nicotine concentration, in
64 samples the nicotine concentration was found
to be as low from -1 to -67.13 % and in remaining
5 samples it contained higher from +2.8 to +24.70
%. A total of 60 (86.9 %) samples were found to
have significant discrepancies in the labelled vs.
actual nicotine concentration that was calculated
by one sample t-test. The estimation of nicotine
analysis revealed the necessity for good manu-
facturing and quality standards of these nicotine-
containing products in Malaysia. The graphical
presentation of the current study is described in
Figure. 2.
Discussion
The study was first of its kind in Malaysia, which
determines the actual nicotine concentration in the
Malaysian marketed e-cig e-liquids brands that
Figure 1. Calibration curve
Azizur Rahman et al., / TACL 8 (1) 2018 54 - 62 56
Table 1. Estimation of labelled vs actual nicotine
concentration among malaysian marketed e-liquids samples
Sample E-Liquids Labelled nicotine Average actual SD (±) % Df P Value
No. samples conc (Mg/Ml) nicotine conc
(Mg /Ml)
1 Sample 01 6.00 3.36 0.4394 -44.05* .014
2 Sample 02 6.00 2.18 0.1032 -63.68* <.001
3 Sample 03 6.00 3.33 0.3967 -44.44* .004
4 Sample 04 6.00 3.85 1.1589 -35.83* .048
5 Sample 05 6.00 3.24 0.0509 -45.93* .008
6 Sample 06 6.00 4.76 0.8485 -20.68 .117
7 Sample 07 6.00 2.78 0.1492 -53.74* .001
8 Sample 08 6.00 2.79 0.3550 -53.55* .003
9 Sample 09 6.00 2.45 0.2715 -59.20* .010
10 Sample 10 6.00 2.41 0.3571 -59.88* .002
11 Sample 11 6.00 7.27 0.0566 +21.17* .028
12 Sample 12 6.00 4.51 0.3111 -24.83 .142
13 Sample 13 6.00 5.94 0.8910 -1.00 .974
14 Sample 14 6.00 2.16 0.1216 -63.97* <.001
15 Sample 15 6.00 2.12 0.0841 -64.61* <.001
16 Sample 16 6.00 2.13 0.0488 -64.58* <.001
17 Sample 17 6.00 2.29 0.4490 -61.88* .002
18 Sample 18 6.00 2.43 0.0849 -59.50* <.001
19 Sample19 6.00 2.28 0.2192 -61.98* <.001
20 Sample 20 6.00 2.07 0.1655 -65.50* <.001
21 Sample 21 6.00 2.00 0.0778 -66.75* <.001
22 Sample 22 6.00 2.03 0.0594 -66.17* .002
23 Sample 23 6.00 2.26 0.3663 -62.37* <.001
24 Sample 24 6.00 2.02 0.0954 -66.27* <.001
25 Sample 25 6.00 2.22 0.1061 -63.02* <.001
26 Sample 26 6.00 2.52 0.1711 -57.95* .001
27 Sample 27 6.00 2.25 0.1400 -62.48* <.001
28 Sample 28 6.00 2.23 0.3041 -62.92* <.001
29 Sample 29 6.00 2.06 0.1464 -65.73* <.001
30 Sample 30 6.00 2.13 0.1902 -64.58* .002
31 Sample 31 6.00 2.97 0.3504 50.46 .002
32 Sample 32 6.00 1.97 0.0537 -67.13* <.001
33 Sample 33 6.00 7.40 1.1370 +23.32 .186
34 Sample 34 18.00 14.25 0.2758 -20.86* 0.003
35 Sample 35 12.00 7.61 0.1054 -36.55* 0.002
36 Sample 36 6.00 4.25 0.0141 -29.17* .009
37 Sample 37 3.00 3.34 0.3550 +11.30 .128
38 Sample 38 3.00 2.79 0.7326 -7.03 .398
39 Sample 39 3.00 2.19 0.2821 -26.98 .356
40 Sample 40 6.00 4.33 0.4285 -27.88* .048
41 Sample 41 6.00 5.03 0.8485 -16.17 .201
Azizur Rahman et al., / TACL 8 (1) 2018 54 - 62 57
table 1. (continued).
Sample E-Liquids Labelled nicotine Average actual SD (±) % Df P Value
No. samples conc (Mg/Ml) nicotine conc
(Mg /Ml)
42 Sample 42 3.00 2.13 0.2348 -28.87 .053
43 Sample 43 3.00 3.08 0.1075 +2.80 .169
44 Sample 44 6.00 4.49 0.0141 -25.17* .001
45 Sample 45 6.00 3.42 1.2869 -43.00* .038
46 Sample 46 6.00 3.22 0.0410 -46.32* .002
47 Sample 47 6.00 4.02 0.2192 -33.08* .003
48 Sample 48 6.00 2.82 0.0778 -53.08* .002
49 Sample 49 6.00 3.97 0.2475 -33.92* .004
50 Sample 50 6.00 4.92 0.0990 -18.00* .011
51 Sample 51 6.00 4.04 0.2333 -32.75* .003
52 Sample 52 6.00 4.55 0.3041 -24.25* .009
53 Sample 53 6.00 2.96 0.0898 -50.61* .004
54 Sample 54 6.00 4.36 0.0495 -27.42* .001
55 Sample 55 6.00 4.35 0.3578 -27.45* .015
56 Sample 56 6.00 4.45 0.2055 -25.89* .006
57 Sample 57 6.00 3.92 0.8485 -34.67* .015
58 Sample 58 6.00 4.12 0.2899 -31.42* .006
59 Sample 59 6.00 4.75 0.0424 -20.83* .038
60 Sample 60 6.00 2.34 0.0849 -61.00* .005
61 Sample 61 6.00 3.61 0.6866 -39.86* .024
62 Sample 62 6.00 2.56 0.6406 -57.28* .012
63 Sample 63 6.00 4.70 0.0849 -21.67* .004
64 Sample 64 6.00 3.49 0.8443 -41.88* .018
65 Sample 65 6.00 3.81 0.2333 -36.58* .008
66 Sample 66 6.00 4.40 0.2828 -26.67* .006
67 Sample 67 3.00 3.20 0.1676 +6.52 .099
68 Sample 68 6.00 4.52 0.0028 -24.70* .003
69 Sample 69 6.00 3.69 0.1322 -38.44* .002
* Indicate significant discrepancies in the labelled vs actual nicotine concentration calculated by one
sample t test; SD = Standard Deviation
% DF = Percentage Difference; CONC = Concentration
were never reported earlier. The chemical analy-
sis found inconsistencies in the labelled vs. actual
nicotine concentration among the Malaysian mar-
keted e-liquids products. The study discovered
high discrepancies in the nicotine concentration
and rose up safety issues related to marketed e-
liquids brands for consumer safety purpose in
Malaysia. The study results revealed a high in-
consistency in the nicotine concentration among
more than 85 % of the marketed e-liquids samples.
Similar to our findings some of the previous stud-
ies from other countries like America, Korea,
Poland, China 8,9,10 were too discovered inconsis-
tencies in the nicotine concentration among their
marketed e-liquids products. The main safety is-
sues related to the theses nicotine-containing prod-
ucts in Malaysia is due to the lack of manufactur-
ing guidelines. Presently, the absence of manda-
Azizur Rahman et al., / TACL 8 (1) 2018 54 - 62 58
Electronic cigarette 69 E-liquid sample from Malaysian market
Percentage and significance difference between
labelled vs actual nicotine concentration
Mass spectrometer
(ion source, quadrupole
mass filter, ion detector)
Nicotine estimation
Sample injection
GC-MS
Figure 2. Graphical presentation of the current study
tory manufacturing guidelines from the tobacco-
controlled board, these e-liquids are not formulat-
ing as per their standards. The study indicated
that the urgent formulations strategies should be
established for both local and international brands
which are available in Malaysia by the agencies
that control the nicotine-containing products.
The current study exposed that the majority of
Azizur Rahman et al., / TACL 8 (1) 2018 54 - 62 59
the e-liquids showed a lower level of nicotine than
labelled claimed by the manufacturer. However,
the low nicotine concentration in the e-liquids may
misinform the consumers and overestimate the
nicotine experience. Additionally, due to lower
concentration, the vapers consider e-cig is less
satisfactory and ineffective substitute for the to-
bacco cigarette. Our study finding is different as
compared to the analysis performed by the Etter
et al. The Etter et al. tested 20 commercial e-
liquids brands from China, US, France, UK, Bel-
gium and found nicotine level in the e-liquids bottles
were nearly similar to the labelled values as
claimed by the manufacturer 14
. However, our
study findings are comparable to the study results
which reported by Davis et al. and Goniewicz et
al. 8-10. The directly above result discussion sug-
gests that the quality of these e-liquids vary from
each country where the products are formulated
and packed.
The existing study identified nicotine concen-
tration of as low as -67.13 % and the highest con-
centration of +24.7 % as compared to the label
claimed by the makers which ranged from 3 mg
to 18 mg/ml. However, the current study did not
recognise any serious threatening level of high
nicotine concentration amount like between 100-
200 mg/ml in Malaysian marketed products which
were identified by Gronkiewicz et al. 8. The high
nicotine concentrated products in e-liquids can in-
duce nicotine overdosing and may upsurge the
threat of accidental poisoning. Therefore, the Eu-
ropean Union directive fixed the limit of maxi-
mum nicotine concentration in ready to use re-
filled e-liquids as 20 mg/ml in a bottle of not more
than 10 ml for the consumer safety reasons 15.
Tobacco experts agreed that e-cig is not totally
hazarded free but less harmful than tobacco smok-
ing 6, 7. The tobacco professionals believed that
the threat of e-cig is from its components, which
include nicotine, propylene glycol or glycerol and
various other flavouring agents. Tobacco analysts
have high concern over the use of nicotine due to
its poisoning and addictive nature. Besides, the
nicotine use is associated with some health risk in
the smokers, teenagers and pregnant women’s.
In addition, there is strong evidence that the nico-
tine use has a role in neurodegeneration and im-
paired brain develop-ment among the consumers
primarily in the youths 16 . Moreover, some acci-
dental and intentional poisoning cases of nicotine
have been reported among the children and adults
in the preceding e-cig related studies 17, 18. Fur-
thermore, WHO does not recommend any nico-
tine-containing products usage due to its extremely
addictive nature 19.
Currently, e-cig is a new phenomenon and ex-
perts considered that the e-cig is a tobacco harm
reduction (THR) device rather than the effec-
tive smoking cessation aid because it consists of
nicotine but without smoke 7, 20, 21. Some advi-
sory groups described the THR strategy can use
in health care and in social policy to reduce dam-
age to the individual or large peoples from the
harmful effects of tobacco use that cannot en-
tirely avoid or prohibited. The aim of the THR
policy is to provide the nicotine to the tobacco
users to which they are addicted but deprived of
smoke 23.
Nevertheless, the significant differences in the
nicotine concentrations indicated the missing stan-
dard and manufacturing guidelines for commer-
cially available nicotine e-liquids amongst the
Malaysian marketed samples. The current study
showed variances in the nicotine concentration in
the e-liquids as per the labelled value. The study
alarmed that the dissimilarity in the nicotine level
may suggest the interaction of nicotine with pack-
aged material or with other e-liquids contents like
propylene glycol, vegetable glycerine or other
flavouring agents. This study also suggests that
the difference in the nicotine concentration may
imply due to improper handling and storage of e-
liquids at the retail site. The current study also
suggests that the e-liquids market in Malaysia has
generally established outside a suitable governing
framework. This inappropriate regulatory struc-
ture permitting the e-liquids makers and whole-
salers to develop a product without knowledge of
safety. Therefore, outcomes from the current
study suggest that the regulation of nicotine-con-
taining e-liquids with accurate labelling and pack-
aging are required immediately for consumer
safety purpose. In Malaysia, a voluntary
organisation, Malaysian e–vaporisers and tobacco
alternative Association (MEVTA) have estab-
Azizur Rahman et al., / TACL 8 (1) 2018 54 - 62 60
lished some manufacturing standards for the e-
liquids. However, no manufacturing guidelines
from the federal government are available for
quality improvements for these nicotine-contain-
ing e-liquids. The current study is not without limi-
tation. The investigator selected the samples based
on a suggestion from prominent e-liquid suppliers
on a regional level. Therefore, due to lack of na-
tionwide marketing data of these e-liquids the in-
vestigator had not capable of confirming whether
the selected brands are popular at the national
level and sold by the retails shops in other states
of Malaysia.
The researcher did not estimate the nicotine con-
centration in across the batches of same brands
of e-liquids. Further, the investigator did not esti-
mate the nicotine concentration in the products
that labelled as nicotine-free due to its non-popu-
larity 24. Finally, the investigator analysed many e-
liquids that labelled claimed as manufactured in
Malaysia but the researcher believed that China
remains the major supplier of e-liquids to the Ma-
laysia and there is a possibility that some of the
tested Malaysian products may import from the
China.
Conclusion
As per the results, the study concludes that there
is a high discrepancy in the labelled vs actual nico-
tine concentration among the Malaysian marketed
e-liquids brands. Based on analysis results, there
is an urgent need for regulation for these mar-
keted available nicotine e-liquids in Malaysia. The
study reveals that the tobacco administrative au-
thority in Malaysia should instruct all e-liquids sup-
pliers and manufacturers nationwide for flawless
labelling and packaging to ensure superior quality
of these nicotine-containing products. Furthermore,
the e-liquids regulation guidelines should enforce
the manufacturers to supply these products in
childproof packaging to minimise the risk of acci-
dental poisoning to the children.
Conflict of interest
All authors declared that they have no conflict
of interest.
Source of funding
The study received fund from the International
Islamic University of Malaysia under Research
Initiative Grant Scheme RIGS NO-15-095-0094.
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