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Forensic chemical profiling of flavouring additives in seized mu’assel (shisha) by gas chromatography-mass spectrometry (GC-MS)

Authors:
  • Directorate of Forensic Science Services, Ministry of Home Affairs, Government of India
  • CFSL Chandigarh

Abstract and Figures

Abstract Background Water-pipe tobacco smoking obsession has again spread worldwide. Especially, the younger generation is besotted with mu’assel (shisha) and avidly use these for smoking. Numbers of additives are being added in commercial brands of mu’assel (shisha) to impart diverse taste with amusing aroma. The name of only one of the masking flavours like strawberry, chocolate, vanilla etc. and not the chemical ingredients are printed on the packets of mu’assel. The manufactures remain silent about mentioning the chemical ingredients used for flavourings as they may not have scientific data about chemical compounds attributing to multiple flavours. There is also a shortfall in quality control owing to non-availability of technical procedure(s) to identify masking multiple flavouring additives. Many of these flavoured additives are either carcinogenic or potentially hazardous for human health. Ignoring health hazards, the avaricious manufacturers are intentionally adding multiple additives to make their products more addictive in order to increase their sales. The need of the hour is to unequivocally establish a technique for chemical profiling of the flavouring additives in mu’assel. In this paper, seven popular commercial brands of mu’assel were extracted, sonicated and analysed by GC-MS technique for detection of flavouring additives. Results Twenty-eight flavouring additives, i.e. camphor, linalool, benzyl ethanol, β-citronellol, menthol, vanillin, ethyl vanillin, eugenol, eucalptol, patchouli alcohol, nerol, rheosmin, musk ambrette, musk ketone, phenyl ethyl methyl ether, anethole, estragole, limonene, benzaldehyde, terpineol, phenyl ethyl butyrate, phenethyl isobutyrate, piperonal, methyl isobutyrate, methyl dihydro jasmonate, anisyl alcohol, trans-geraniol and sabinene along with nicotine were detected in varied proportions by GC-MS technique in seven seized popular commercial brands of mu’assel. Conclusion A study on chemical profiling of flavouring additives in commercial mu’assel has yet not been reported. Henceforth, this forensic attempt was aimed to secure public health by chemically profiling the flavouring additives of mu’assel. Many of the detected additives may cause severe health problems. Moreover, the smoker may suffer from neuroticism and psychoticism that may lead to a number of cases pertaining to physical assaults and sexual harassment.
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O R I G I N A L A R T I C L E Open Access
Forensic chemical profiling of flavouring
additives in seized muassel (shisha) by
gas chromatography-mass spectrometry
(GC-MS)
Deepak Middha and Archna Negi
*
Abstract
Background: Water-pipe tobacco smoking obsession has again spread worldwide. Especially, the younger generation is
besotted with muassel (shisha) and avidly use these for smoking. Numbers of additives are being added in commercial
brands of muassel (shisha) to impart diverse taste with amusing aroma. The name of only one of the masking flavours
like strawberry, chocolate, vanilla etc. and not the chemical ingredients are printed on the packets of muassel. The
manufactures remain silent about mentioning the chemical ingredients used for flavourings as they may not have
scientific data about chemical compounds attributing to multiple flavours. There is also a shortfall in quality control owing
to non-availability of technical procedure(s) to identify masking multiple flavouring additives. Many of these flavoured
additives are either carcinogenic or potentially hazardous for human health. Ignoring health hazards, the avaricious
manufacturers are intentionally adding multiple additives to make their products more addictive in order to increase their
sales. The need of the hour is to unequivocally establish a technique for chemical profiling of the flavouring additives in
muassel. In this paper, seven popular commercial brands of muassel were extracted, sonicated and analysed by GC-MS
technique for detection of flavouring additives.
Results: Twenty-eight flavouring additives, i.e. camphor, linalool, benzyl ethanol, β-citronellol, menthol, vanillin, ethyl
vanillin, eugenol, eucalptol, patchouli alcohol, nerol, rheosmin, musk ambrette, musk ketone, phenyl ethyl methyl ether,
anethole, estragole, limonene, benzaldehyde, terpineol, phenyl ethyl butyrate, phenethyl isobutyrate, piperonal, methyl
isobutyrate, methyl dihydro jasmonate, anisyl alcohol, trans-geraniol and sabinene along with nicotine were detected in
varied proportions by GC-MS technique in seven seized popular commercial brands of muassel.
Conclusion: A study on chemical profiling of flavouring additives in commercial muassel has yet not been reported.
Henceforth, this forensic attempt was aimed to secure public health by chemically profiling the flavouring additives of
muassel. Many of the detected additives may cause severe health problems. Moreover, the smoker may suffer from
neuroticism and psychoticism that may lead to a number of cases pertaining to physical assaults and sexual harassment.
Keywords: Forensic chemistry, Carcinogenic, Flavour, GC-MS, Nicotine, Smoking, Muassel, Hookah, Psychoticism,
Public health
© The Author(s). 2019 Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and
reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to
the Creative Commons license, and indicate if changes were made.
* Correspondence: archnaforensic@yahoo.com
Central Forensic Science Laboratory, Directorate of Forensic Science Services,
Ministry of Home Affairs, Govt. of India, Plot No. 2, Dakshin Marg, Sector 36
A, Chandigarh 160036, India
Egyptian Journal o
f
Forensic Science
s
Middha and Negi Egyptian Journal of Forensic Sciences (2019) 9:39
https://doi.org/10.1186/s41935-019-0146-2
Background
Tobacco is one of the most widely abused substances
in the world and is found to be highly addictive for its
major alkaloid stimulantnicotine (Ralapati and Lu
1998). The tobacco plant is a part of the genus Nicoti-
ana and of the Solanaceae (nightshade) family. While
more than 70 species of tobacco are known, the chief
commercial crop is Nicotiana Tabacum (Fig. 1).
Dried cured tobacco leaves are consumed by various
ways such as in cigarettes,cigars,muassel, flavoured
tobacco,snuff,chewing tobacco,dipping tobacco and
snus. Among these, the most popular way to consume
tobacco is muassel through a hookah or water pipe
(Fig. 2) in which the vapour or smoke of muassel is
passed through a water basin before inhalation (Wiki-
pedia 2015). Muassel or shisha is becoming increas-
ingly popular due to its flavoured tobacco smoke. It is
a syrupy tobacco mix with molasses and vegetable gly-
cerol as moisturiser. Some specific flavours are also
added to it. Typical flavours of muassel include straw-
berry, chocolate, vanilla, apple, grape, guava, lemon
and mint, as well as many other fruit-based mixtures.
In this form, the user gets nicotine along with multi-
flavour smoke.
Additives are used primarily for flavours and smooth-
ing the smokers experience (Rabinoff et al. 2007).
Ninety-five percent of chemicals used in fragrances are
synthetic compounds derived from petroleum. Smoking
them can cause cancer, birth defects, CNS disorders and
allergic reactions, e.g. limonenecarcinogenic, benzyl al-
coholCNS depressants, camphorCNS stimulant and
linaloolCNS depressants (EPA 2015).
Some of these chemicals are also found addictive,
e.g. menthol increases nicotine receptor density (Cal-
lier V 2014). Henceforth, the Tobacco Products Dir-
ective (2001/37/EC) was provided for EU member
states to regulate tobacco products so as to protect
public health. This directive included the prohibition
of ingredients which increase the addictiveness of to-
bacco products (Europa 2015). Owing to the short falls
in quality control norms and non-availability of a reli-
able facility to identify flavouring additives, avaricious
manufacturers are intentionally adding additives with-
out considering the adverse effect of these chemicals.
Moreover, these manufacturers are only printing the
name of the masking flavour and not the details of the
other additives on the cover of the commercial muas-
sel. Due to these unfair practices, smokers remain un-
aware about the health hazard of flavouring additives
present in muassel. Some methods were developed for
the detection of nicotine from tobacco leaves by GC-
MS (Hossain AM et al. 2013)and for detection of
furanic compounds in the smoke of the water pipe
(Schubertetal.2011).
Study on chemical profiling of flavouring additives
in muassel has not yet been reported. Henceforth, a
Fig. 1 Nicotiana Tabacum (Wikipedia 2015)
Middha and Negi Egyptian Journal of Forensic Sciences (2019) 9:39 Page 2 of 13
forensicattemptismadetosecurepublichealthby
chemically profiling flavouring additives in muassel.
Forensic study was mainly targeted to give complete
forensic chemical profiling of muasselsoastomake
smokers aware of the hidden health hazards of these
products. In this study, representative samples from a
large number of commercial varieties of muassel were
subjected to forensic chemical analysis. These com-
mercial varieties of muassel were seized during raids
conducted in different hookah bars in Chandigarh by
Chandigarh Police under COPTA Act, 2003, and were
submitted at the Central Forensic Science Laboratory,
Chandigarh, for the detection of nicotine. The samples
of commercial varieties of muassel were ultrasoni-
cated (Verma and Middha 2010) and analysed by using
gas chromatography-mass spectrometry (GC-MS)
technique. Twenty-eight flavouring additives apart
from nicotine were identified by the mass spectral
chemical profiling.
Materials and method
Representative varieties and reagents
The representative samples were taken from seven
commercial varieties of muassel seized by Chandigarh
police and were marked as R.V-1 to R.V-7.
Variety Brand Representative samples
1 Aladdin Lady DragonR.V-1
2 Aladdin Sweet 16R.V-2
3 Aladdin Brain FreezerR.V-3
4 Aladdin X on BeachR.V-4
5 Qehwa Pan SalsaR.V-5
6 MyaBombay Blue Pan R.V-6
7 Mya Pan RasnaR.V-7
The packets of all the seized varieties of muassel were
printed with only details of nicotine and tar, whereas the
details of other multiple flavouring additives were not
mentioned on any of the packets of muassel.
The solvent used for extraction was of LC grade
(Merck, German).
Equipment
Ultrasonicator of PCi, Mumbai, India; Balance-AB104S
of Mettler, Toledo, Switzerland; Thermo Finnigan Trace
GC Ultra coupled with a Thermo DSQ Quadrupole MS
and Thermo autosampler 3000 were used (directive
2001/37/EC).
Fig. 2 Hookah (Wikipedia 2015)
Middha and Negi Egyptian Journal of Forensic Sciences (2019) 9:39 Page 3 of 13
Ultrasonic extractions of muassel for flavouring additives
Ten millilitres of chloroform was added to a 2-g repre-
sentative sample of each variety in a round bottom
flask and kept for 8 h in a refrigerator. These were
then ultrasonicated for 1 followed by filtration
through Whatman No.1 filter paper (GE Healthcare,
UK). The filtrated samples were concentrated to 2 ml
andstoredat4°C.
Instrumentation conditions
A Thermo Finnigan Trace GC Ultra coupled with a
Thermo DSQ Quadrupole MS and Thermo autosampler
3000 was used. The column was a 30-m BP-5 with 0.3-
mm I.D. and 0.5-μm film thickness. Helium was used as
a carrier gas at a constant flow of 1.2 ml/min. Splitless
injection was used with a splitless time of 60 s. The in-
jector and interface line temperature were held at 250 °C
and 330 °C respectively. Oven temperature was held at
90 °C for 1 min and increased to 310 °C at the rate of
20 °CC/min and held at this temperature for 10 min.
The MSD conditions: ionisation energy 70 eV, ion
source temperature 200 °C, mass range 41410 amu,
electron multiplier voltage (Auto tune + 200 V).
Sample injection volume: 1 μl
Compound identification
Xcaliber 1.4 software was used for data acquisition and
processing and results were screened using the library
of National Institute of Standard and Technology.
Results
Various chemicals apart from nicotine were detected in
R.V-1 to R.V-7 by GC-MS technique. Out of these, 28
flavouring additives/ chemicals were found responsible
for flavour in these representative varieties. These 28
flavouring chemicals are tabulated against their respect-
ive detected retention time (R
t)
in Tables 1,2,3,4,5,6
and 7and their resulting total ion chromatogram are
depicted in Figs. 3,4,5,6,7,8and 9respectively. Com-
bined chemical profiling of these 28 flavouring additives
Table 1 Detected flavouring additives in R.V-1
R
t
Chemical
compound
Molecular
weight
(g/mol)
Important fragments
5.54 Limonene 136.24 68, 67, 93, 53, 94, 136
6.30 Linalool 154.25 71, 93, 55 ,121 ,136 ,139
6.51 Benzene ethanol 108.14 91, 92, 122, 65, 39, 123
6.91 camphor 152.24 95, 81, 69, 108, 152, 153
7.16 Menthol 156.27 71, 81, 95, 55, 123, 138
7.65 β-Citronellol 156.27 41, 69 ,81 ,82 ,123 ,138
7.93 Nerol 154.25 41, 69, 93, 123, 136, 154
8.31 Anethole 148.21 148, 147, 133, 77, 51, 149
8.96 Nicotine 162.2 84, 45, 133, 162, 55, 119
8.98 Eugenol 164.20 164, 149, 77, 55, 103, 165
9.40 Vanillin 152.15 151, 152, 81, 123, 53, 153
9.89 Ethyl Vanillin 166.18 137, 166, 109, 81, 29, 167
10.66 Rheosmin 164.20 107, 164, 43, 121, 94, 165
11.75 Patchouli alcohol 222.36 83, 138, 98, 222, 81, 55
Table 2 Detected flavouring additives in R.V-2
R
t
Chemical
compound
Molecular
weight (g/mol)
Important fragments
5.58 Eucalptol 154.249 81, 71, 108, 69, 111, 154
6.16 Phenyl ethyl methyl
ether (Kewda ether)
136.19 45, 91, 136, 104, 65, 137
6.53 Benzeneethanol 108.14 91, 92, 65, 122, 39, 123
6.93 Camphor 152.24 95, 81, 69, 108, 41, 152
7.16 Menthol 156.27 71, 81, 95, 41, 123, 138
7.65 β-Citronellol 156.27 41, 69, 81, 82, 123, 138
7.98 Estragole 148.20 148, 147, 77, 121, 91, 51
8.33 Anethole 148.21 148, 147, 117, 77, 51, 149
8.84 Piperonal 150.13 149, 150, 63, 121, 65, 62
8.95 Nicotine 162.2 84, 133,162,42,119,65
8.98 Eugenol 164.20 164, 149, 77, 103, 55, 165
9.42 Vanillin 152.15 151, 152, 81, 123, 153, 154
9.89 Ethyl vanillin 166.18 137, 166, 109, 81, 29, 167
11.75 Patchouli alcohol 222.36 83, 138, 98, 222, 81, 55
12.91 Musk ambrette 268.26 253, 43, 91, 77, 268, 115
Table 3 Detected flavoured additives in R.V-3
R
t
Chemical
compound
Molecular weight
(g/mol)
Important fragments
5.54 Limonene 136.24 68, 67, 93, 94, 136, 137
5.86 Sabinene 136.23 93, 91, 71, 41, 136, 137
6.30 Linalool 154.25 71, 93, 43, 121, 136, 139
6.51 Benzene ethanol 108.14 91, 92, 65, 122, 39, 123
6.91 Camphor 152.24 95, 81, 108, 41, 152, 55
7.16 Menthol 156.27 71, 81, 95, 55, 123, 138
7.35 Terpineol 154.25 59, 93, 121, 136, 81, 139
7.65 β-Citronellol 156.27 41, 69, 81, 82, 123, 138
7.91 Trans-gerniol 154.25 69, 41, 68, 93, 123, 139
8.95 Nicotine 162.2 84, 133, 162, 42, 119, 65
8.98 Eugenol 164.20 164, 149, 77, 103, 55, 39
9.40 Vanillin 152.15 151, 152, 81, 123, 53, 153
9.72 Phenyl ethyl
butyrate
192.25 104, 105, 71, 91, 106, 161
9.91 Ethyl vanillin 166.18 137, 138, 166, 109, 81, 167
10.66 Rheosmin 164.20 107, 43, 164, 77, 121, 165
12.91 Musk ambrette 268.26 253, 43, 91, 77, 268, 115
13.93 Musk ketone 294.30 43, 279, 128, 294, 115, 129
Middha and Negi Egyptian Journal of Forensic Sciences (2019) 9:39 Page 4 of 13
detected in R.V-1 to R.V-7 are summarised in Table 8
and the flavour of each detected chemical along with its
chemical structure is summarised in Table 9.
Discussion
Twenty-eight flavouring additives were detected in
seven representative varieties of muassel by GC-MS
technique summarised in Tables 1,2,3,4,5,6,,7and
8 and TICs of R.V-1 to R.V-7 are depicted in Figs. 3,4,
5,6,7,8and 9respectively. Some of these detected
chemicals are derived from plants origin, some are
synthesised and some are both: These are categorised
as under:
Table 5 Detected flavouring additives in R.V-5
R
t
Chemical
compound
Molecular
weight
(g/mol)
Important fragments
5.60 Eucalptol 154.249 81, 71, 108, 111, 154, 69
6.16 Phenyl ethyl methyl
ether (Kewda ether)
136.19 91, 136, 45, 65, 104, 137
6.53 Benzene ethanol 108.14 91, 92, 65, 122, 64, 123
7.16 Menthol 156.27 l 71, 81, 95, 123, 138, 67
7.65 β-Citronellol 156.27 41, 69, 81, 82, 123, 138
7.93 Trans-gerniol 154.25 69, 41, 93, 123, 139, 154
8.96 Nicotine 162.2 84, 133, 162, 42, 119, 163
9.40 Vanillin 152.15 151, 152, 153, 81, 123, 53
11.75 Patchouli alcohol 222.36 83, 138, 98, 222, 81, 55
12.91 Musk ambrette 268.2 253, 43, 91, 268, 77, 115
Table 6 Detected flavouring additives in R.V-6
R
t
Chemical
compound
Molecular
weight
(g/mol)
Important fragments
4.75 Benzaldehyde 106.12 105, 106, 77, 78, 107
5.60 Eucalptol 154.249 43, 81, 71, 108, 111, 154
6.17 Phenyl ethyl methyl ether 136.19 91, 136, 45, 65, 104, 103
6.30 Linalool 154.25 71, 93, 55, 69, 121, 136
6.53 Benzene ethanol 108.14 91, 92, 122, 65, 39, 93
6.93 Camphor 152.24 95, 81, 69, 108, 41, 152
7.16 Menthol 156.27 71, 81, 95, 82, 123, 138
7.65 β-Citronellol 156.27 41, 69, 81, 82, 123, 138
7.93 Trans-gernaniol 154.25 69, 41, 68, 93, 123, 139
8.28 Anisyl alcohol 138.17 138, 137, 109, 77, 94, 39
8.96 Nicotine 162.2 84, 133, 162, 42, 119, 163
8.98 Eugenol 164.20 164, 149, 77, 103, 131, 55
9.40 Vanillin 152.15 151, 152, 81, 109, 53, 50
9.89 Ethyl vanillin 166.18 137, 166, 138, 109, 81, 167
10.66 Rheosmin 164.20 107, 164, 43, 77, 121, 65
11.75 Patchouli alcohol 222.36 83, 138, 98, 222, 81, 55
Table 7 Detected flavouring additives in R.V-7
R
t
Chemical
compound
Molecular
weight
(g/mol)
Important fragments
5.60 Eucalptol 154.249 43, 81, 108, 71, 111, 154
6.16 Phenylethylmethylether
(Kewda ether)
136.19 45, 91, 136, 104, 65, 137
6.30 Linalool 154.25 71, 93, 55, 68, 121, 136
6.53 Benzene ethanol 108.14 91, 92, 122, 65, 39, 123
6.93 Camphor 152.24 95, 81, 69, 108, 41, 152
7.17 Menthol 156.27 71, 81, 95, 55, 123, 138
7.65 β-Citronellol 156.27 41, 69, 81, 82, 123, 138
7.91 Trans-geraniol 154.25 69, 41, 93, 123, 139, 154
8.31 Estragole 148.20 148, 147, 77, 121, 91, 51
8.95 Nicotine 162.2 84, 133, 162, 82, 92, 163
8.98 Eugenol 164.20 164, 149, 77, 103, 55, 165
9.40 Vanillin 152.15 151, 152, 81, 123, 53, 153
11.75 Patchouli alcohol 222.36 41, 43, 83, 98, 138, 222
12.91 Musk ambrette 268.26 253, 268, 43, 91, 77, 115
13.93 Musk Ketone 294.30 43, 279, 128, 294, 115, 129
Table 4 Detected flavouring additives in R.V-4
R
t
Chemical
compound
Molecular
weight (g/mol)
Important fragments
5.60 Eucalptol 154.249 43, 81, 71, 108, 111, 154
5.81 Methyl isobutyrate 102.13 43, 40, 71, 87, 102, 103
6.16 Phenyl ethyl
methyl ether
136.19 91, 136, 104, 65, 137, 138
6.30 Linalool 154.25 71, 43, 93, 55, 121, 136
6.51 Benzene ethanol 108.14 91, 92, 65, 122, 39, 123
6.93 Camphor 152.24 95, 81, 41, 108, 109, 152
7.16 Menthol 156.27 71, 81, 95, 55, 67, 123
7.65 β-Citronellol 156.27 41, 69, 81, 82, 123, 138
7.93 Nerol 154.25 41, 69, 68, 93, 123, 136
8.31 Anethole 148.21 148, 147, 117, 77, 105, 51
8.96 Nicotine 162.2 84, 133, 162, 92, 82, 163
8.98 Eugenol 164.20 164, 149, 77, 103, 55, 165
9.40 Vanillin 152.15 151, 152, 81, 123, 53, 153
9.74 Phenethyl
isobutyrate
192.254 104, 43, 71, 105, 106, 42
10.66 Rheosmin 164.20 107, 43, 164, 77, 121, 65
11.45 Methyl dihydro
jasmonate
226.32 83, 156, 153, 82, 55, 96
11.73 Patchouli alcohol 222.36 41, 43, 83, 98, 138, 222
12.89 Musk ambrette 268.2 253, 268, 91, 77, 115, 145
13.93 Musk ketone 294.30 43, 279, 128, 294, 115, 129
Middha and Negi Egyptian Journal of Forensic Sciences (2019) 9:39 Page 5 of 13
Fig. 3 Total ion chromatogram of R.V-1
Fig. 4 Total ion chromatogram of R.V-2
Middha and Negi Egyptian Journal of Forensic Sciences (2019) 9:39 Page 6 of 13
1. Plant origin: camphor, linalool, benzyl alcohol, β-
citronellol, menthol, vanillin, eugenol, patchouli
alcohol, nerol, eucalptol, rheosmin, anethole,
estragole, limonene, benzaldehyde, terpineol,
trans-geraniol and sabinene
2. Synthetic: musk ambrette, musk ketone, ethyl
vanillin, methyl isobutyrate, phenyl ethyl butyrate,
phenethyl isobutyrate, phenyl ethyl methyl ether
and methyl dihydro jasmonate
3. Both (plant origin as well as synthetic): piperonal
and anisyl alcohol
These chemicals are added for imparting flavour into
the muassel by the manufacturers. These detected
chemicals owe specific properties for impartinga
characteristic flavour. The flavour of each detected
chemical is summarised in Table 9.Someofthese
chemicals impart similar flavours. The manufacturers
inadvertently added on multiple chemicals imparting
thesameflavourinmuassel, i.e for vanilla flavour
vanillin and ethyl vanillin in R.V-1, R.V-2, R.V-3 and
R.V-6; for spicy flavoureugenol and eucalptol in R.V-
2, R.V-4, R.V-6 and R.V-7 and for musk aromamusk
ambrette and musk ketone in R.V-3, R.V-4 and R.V-7
and for aroma of mixture of citrus and floralβ-
citronellol and methyl dihydro jasmonate in R.V-4.
TICs of seven representative varieties are depicted in
Figs. 3,4,5,6,7,8and 9. Menthol was computed to
have high proportion in R.V-1, R.V-3, R.V-5, R.V-6 and
R.V-7, whereas anethole was in high proportion in R.V-
2 and R.V-4. Patchouli alcohol, benzene ethanol and
camphor were detected in R.V-1, R.V-4, R.V-6 and R.V-
7; eucalptol in R.V-2 and R.V-4 to R.V-7; linalool in
R.V-1, R.V-3, R.V-4, R.V-6 and R.V-7 and musk
Fig. 5 Total ion chromatogram of R.V-3
Middha and Negi Egyptian Journal of Forensic Sciences (2019) 9:39 Page 7 of 13
Fig. 6 Total ion chromatogram of R.V-4
Fig. 7 Total ion chromatogram of R.V-5
Middha and Negi Egyptian Journal of Forensic Sciences (2019) 9:39 Page 8 of 13
Fig. 8 Total ion chromatogram of R.V-6
Fig. 9 Total ion chromatogram of R.V-7
Middha and Negi Egyptian Journal of Forensic Sciences (2019) 9:39 Page 9 of 13
Table 8 Combined chemical profiling of detected flavouring additives
S.No. R.V-1 R.V-2 R.V-3 R.V-4 R.V-5 R.V-6 R.V-7
1 Camphor Camphor Camphor Camphor Camphor Camphor
2 Linalool Linalool Linalool Linalool Linalool
3 Benzene
ethanol
Benzene ethanol Benzene
ethanol
Benzene ethanol Benzene ethanol Benzene ethanol Benzene ethanol
4β-Citronellol β-Citronellol β-Citronellol β-Citronellol β-Citronellol β-Citronellol β-Citronellol
5 Menthol Menthol Menthol Menthol Menthol Menthol Menthol
6 Vanillin Vanillin Vanillin Vanillin Vanillin Vanillin Vanillin
7 Ethyl vanillin Ethyl vanillin Ethyl vanillin ––Ethyl vanillin
8 Eugenol Eugenol Eugenol Eugenol Eugenol Eugenol
9Eucalptol Eucalptol Eucalptol Eucalptol Eucalptol
10 Patchouli
alcohol
Patchouli alcohol Patchouli alcohol Patchouli alcohol Patchouli alcohol Patchouli alcohol
11 Nerol ––Nerol –––
12 Rheosmin Rheosmin Rheosmin Rheosmin
13 Musk ambrette Musk ambrette Musk ambrette Musk ambrette Musk ambrette
14 –– Musk ketone Musk ketone ––Musk ketone
15 Phenyl ethyl methyl ether
(Kewda ether)
Phenyl ethyl methyl ether
(Kewda ether)
Phenyl ethyl methyl ether
(Kewda ether)
Phenyl ethyl methyl ether
(Kewda ether)
Phenyl ethyl methyl ether
(Kewda ether)
16 Anethole Anethole Anethole –––
17 Estragole –––Estragole
18 Limonene Limonene ––––
19 –– – – Benzaldehyde
20 –– Terpineol ––––
21 –– Phenyl ethyl
butyrate
––––
22 –– Phenethyl isobutyrate –––
23 Piperonal ––––
24 –– Methyl iso Butyrate ––
25 –– Methyl dihydro jasmonate –––
26 –– – – Anisyl alcohol
27 –– Trans-geraniol Trans-geraniol Trans-geraniol Trans-geraniol
28 –– Sabinene ––––
Middha and Negi Egyptian Journal of Forensic Sciences (2019) 9:39 Page 10 of 13
Table 9 Chemicals detected in R.V-1 to R.V-7 imparting the flavour
Middha and Negi Egyptian Journal of Forensic Sciences (2019) 9:39 Page 11 of 13
ambrette in R.V-2 to R.V-5 and R.V-7. β-citronellol,
menthol, vanillin and benzene ethanol were detected in
all seven representative varieties of muassel.
The collective flavour of these chemicals in seven
representative varieties of muassel are summarised
in Table 10.
For imparting higher flavour notes, the avaricious
manufacturers add multiple chemicals un-proportionally
to muassel without considering their hazardous conse-
quences. Moreover, to increase the sales of their prod-
ucts, manufacturers have been adding multiple chemical
compounds having alike flavour. Owing to these, the fu-
sion of flavours of these multiple chemical compounds
in muassel does not have any specific pleasant flavour
due to interference of flavours of various chemicals.
Conclusion
Twenty-eight chemicals responsible for flavour were
detected in seven representative varieties of muassel.
The presence of these flavoured additives varies in
each of the seven representative varieties. A difference
in the flavoured additives causes a difference in the
flavour of these samples. These flavoured additives
not only add flavour to these muassel products but
alsomakethepersonmoreaddictiveforitstaste.
Moreover, buyers are not aware of these chemicals
andtheirharmfuleffectsasthedetailsofthese
flavoured chemicals are not printed on covers of any
of these representative varieties. Therefore, the
manufacturer should print the name of all the added
chemicals on covers of muassel packets.
The users while using hookah inhale nicotine along
with these chemicals. Some of the chemicals along with
nicotine are accumulated inthe human body while some
are converted to new compounds during the heating
process and also get accumulated in the human body.
These chemicals and their new converted chemicals are
harmful to the body. Some of these chemicals have been
approved as additives to food but these are not tested by
burning. The aforementioned health hazards need
utmost attention by the public authorities; on our part,
we have indeed carried out an in-depth analysis on the
burnt flavouring additives of Muassel vis-à-vis health is-
sues. The findings are being critically reviewed before
making in public.
Forensic significance
Production and consumption of flavoured muassel
are based on various economic, social, cultural and
government policies. Due to loopholes in the
government policy, they are commercially available in
plenty in the market/online purchase and especially in
hookah bars. The consumption of hookah along with
flavoured additives is a major issue related to health
as these are so hazardous that their exposure to the
human body can cause inflammation to monocytes
and can cause severe health problems. Some of these
are neurotoxic and some are even carcinogenic.
Another aspect of the flavoured muassel is the
increasing mortalities due to the disease caused by
smoking hookah and by consumption of flavoured
additives.
The hookah smokers tend to be more extroverted,
tensed, impulsive, depressive, anxious and suffered
from traits of neuroticism and psychoticism which
results in a number of cases pertaining to physical
assault and sexual harassment. To make the situation
worse, there is no internationally accepted standard
Table 10 Collective flavour (taste and aroma) by flavouring additives in muassel varieties
Representative
sample
Collective taste Collective aroma
R.V-1 Cool mint, vanilla, anise, fennel, citrus-rosy with
spiciness
Strong penetrating odour of orange, anise, fennel, sweet rose, raspberry and
patchouli
R.V-2 Cool mint, citrus-rosy, vanilla, licorice with
spiciness
Strong penetrating odour of musk, green floral jasmine metallic fresh rose note,
anise and fennel anisic type odour.
R.V-3 Cool mint, citrus-rosy, vanilla, woody with
spiciness of black pepper
Strong penetrating odour of musk, vanilla, raspberry, orange, lilac and rose
R.V-4 Cool mint, citrus-rosy, vanilla with spiciness Strong penetrating odour of patchouli, rose, spicy, raspberry, musk, green floral
jasmine metallic fresh rose note, anise, fennel, fruity odour of apple or pineapple,
sweet fruity-rose honey floral odour, mixture of floral and citrus
R.V-5 Cool mint, vanilla with spiciness Citrus-rosy, patchouli, spicy, musk, green floral jasmine metallic fresh rose note
R.V-6 Cool mint, vanilla, spicy, rose-like floral Strong penetrating odour of citrus-rosy, patchouli, spicy, vanilla, fruity odour of
raspberry, green floral jasmine metallic fresh rose note and almond odour-like
hawthorn
R.V-7 Floral, cool mint, vanilla, spicy, musk, licorice
type and rose-like floral
Strong penetrating, floral with touch of spiciness, citrus-rosy, vanilla, spicy,
patchouli, musk, green floral jasmine metallic fresh rose note and anisic-type odour
Middha and Negi Egyptian Journal of Forensic Sciences (2019) 9:39 Page 12 of 13
method for analysing flavoured muassel in forensic
cases along with any type of database.
This research publication does have a societical
outreach as it makes the public at large aware about
the hazardous effects of flavouring additives in a
muassel and will aid the public authority to make a
smoke-free world.
Abbreviations
CNS: Central nervous system; COPTA: Cigarette and Other Tobacco Product
Act; EC: European community; EU: European Union; GC-MS: Gas
chromatography-mass spectrometry; LC: Liquid chromatography; MSD: Mass
selective detector; R.V: Representative variety; TIC: Total ion chromatogram;
WHO: World Health Organization
Acknowledgements
The authors express their gratitude to the researchers engaged in restoring
public health and safety and to the public authorities striving to make a
smoke-free world.
The authors are thankful to the Chief Forensic Scientist, DFSS, MHA, Govt. of
India, and Director, CFSL, DFSS, MHA, Govt. of India, Chandigarh, for constant
scientific support and encouragements.
Authorscontributions
Dr. DM has planned, reviewed, guided the research experimentations,
interpretation and publication frame work. Mrs. AN has participated in
methodology development, experimentations, interpretation and wrote the
manuscript. Both authors have read and approved the final manuscript.
Authorsinformation
Dr. Deepak Middha M.Sc., Ph.D. (F.S), LL.B. (Professional)
Working as Deputy Director & Scientist-D in Chemistry division of Central Fo-
rensic Science Laboratory, Directorate of Forensic Science Services, Ministry
of Home Affairs, Govt. of India, Chandigarh, India. He has brilliantly served for
more than 29 years by making finest real-time applications of multi-scientific
disciplines for the purpose of Criminal Justice. He has about three decades
of professional experience and expertise in forensic chemical examinations
and reporting on evidential clues pertaining to multi-type crimes involving
bombing investigations, explosives, narcotic drugs, psychotropic substances,
liquor, wildlife, bribery, homicide, arson, theft, burglary etc. He has success-
fully deposed hundreds of expert evidences in different Honble Court of
Law. He had the specialisation on the different mass-spectrometric tech-
niques. He has contributed his services in CWs-MS interpretation (United Na-
tions) multiple assignments. He has also made international contributions in
the research and development sector.
Mrs. Archna Negi M.Sc(F.S), PGDCA
Serving in the same organisation as Senior Scientific Assistant in Chemistry
Division. She is persuasive and contributing researcher and having more
than eight years of experience in forensic chemical examinations on
evidential clues pertaining to multi-type crimes involving, narcotic drugs,
psychotropic substances, bribery, liquor, arson etc. She has also experience in
examination of questioned documents pertaining to various problems in
handwriting, typewriting, decipherment of altered and obliterated docu-
ments, printed matters including security documents such as currency notes
and passport.
Funding
The authors received no specific funding for this work
Availability of data and materials
Related literature is available to authors
Ethics approval and consent to participate
Not applicable.
Consent for publication
Not applicable.
Competing interests
The authors declare that they have no competing interests.
Received: 10 April 2019 Accepted: 26 June 2019
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Middha and Negi Egyptian Journal of Forensic Sciences (2019) 9:39 Page 13 of 13
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Menthol increases nicotine addiction by tweaking brain
  • V Callier
Callier V (2014) Menthol increases nicotine addiction by tweaking brain. https:// www.newscientist.com/article/dn26668-menthol-increase-nicotine-addictionby-tweaking-brain. Accessed 10 Dec 2015
  • R S Verma
  • D Middha
Verma R.S, Middha D (2010) Chroma 71:117. https://doi.org/10.1365/s1033700913 98
  • Nicotiana
Nicotiana. https://en.wikipedia.org/wiki/Nicotiana#/media/File:Tabak_9290019.JPG. Accessed 13 Dec 2015