Post on 16-Oct-2019
OBJECTIVES
Objectives
• General objective
The general aim of the present study was to confirm whether there are potential illicit drugs in a popular buccal tobacco brand marketed in Yemen or not.
Specific objectives
1- To determine the source of risk, if any. In the buccal tobacco brand.
2- To identify nicotine in the tested buccal tobacco brand.
3- To quantify the amount of nicotine in a single dose of the buccal tobacco brand.
INTRODUCTION
Tobacco
• Tobacco is the dried leaves of Nicotina species ( family Solanaceae)
• Tobacco products are administered either as inhalation of smoke or by buccal route.
Tobacco • Risks of tobacco
• WHO: tobacco causes heart diseases (heart attack, lung diseases (COPD) and cancer (lung cancer, mouth and larynx cancer)
DEATH
• WHO: tobacco caused 5.4 million deaths in
(2004) and 100 million deaths over the course of
the 20th century
Tobacco • Risks of tobacco
Poisonous compounds
• Carbon monoxide
• Cyanide
• Carcinogens: Tar , benzopyrene, formaldehyde,
cadmium, nickel, arsenic, nitrosamines, and phenols
The alkaloid nicotine is a the cause of dependence
Tobacco • Risks of tobacco
NICOTINE
• Alkaloid
• CNS stimulant and potent parasympathomimetic
• Act as a receptor agonist at nicotinic acetylcholine receptors
• At low dose (21 mg) used for smoking cessation (transdermal patch)
Tobacco • Risks of tobacco
NICOTINE
• Average amount is ≤ 19.2 mg / 1 g of tobacco (1 cigarette) .
• The usual cigarette consumption is 1 packets (20 cigarettes) provide a daily dose of 384 mg )
• Overdose : > 384 mg /day
changes from stimulant to sedative with increasing dosages, a phenomenon known as "Nesbitt's paradox" reducing neuronal activity
• lethal dose 500-1000 mg
ILLICIT NARCOTICS • Drugs that Can cause addiction.
• 2 categories
- Illegal : prohibited to produce, transport or sell globally e.g. cocaine, heroine, etc
-Legal but controlled to produce, transport , sell can be used as medications e.g. morphine, pethidine, methadone, diazepam.
ILLICIT NARCOTICS UNODC (United nations office on drug and Crimes) classification
1. Cannabis : Mariguana, Hashish
2. Opium (raw opium) and opiates e.g. morphine, Heroin
3. Opoids: e.g. methadone
4. Coca (leaves) and Cocaine
5. Amphetamines
6. CNS depressants: Benzodiazepines, barbiturates, methaqualone
7. Hallucinogens e.g. LSD, phencyclidine
RESEARCH
METHODOLOGY
MATERIALS
Materials
• A brand of buccal tobacco ( Hot Filter Khani®, ingredients : tobacco, spices)
• A popular cigarette brand ( Manchester®, JSS tobacco Ltd, UK)
• References standards : nicotine , lidocaine
codeine , ephedrine and caffeine .
• Chemicals : were gift from Yemeni-Egyptian Pharma Co.
1- Analysis of Illicit narcotics
A- TLC screening
TLC Screening was carried out according to TLC systems recommended by UNODC (United
Nations Office on Drugs and Crime).
The tank TLC : (50 cm x 20 cm) was glass and lined with adsorbent paper.
TLC plates (dimensions: 20 x 10 cm; thickness: 0.25 mm)
A- TLC screening Test No Mobile phase Illicit narcotics Reference standards
1 Methanol: conc. ammonia (100:1.5) .
opium alkaloids , heroin, amphetamines, cocaine a, methaqualone and diazepam .
ephedrine, lidocaine
2 Ethyl acetate: methanol: Conc. Ammonia (85:10:5)
opium alkaloids , heroin and amphetamines
Codeine, ephedrine
3 Cyclohexane: toluene: diethylamine (75:15:10).
amphetamines, cocaine, methaqualone and diazepam.
lidocaine, caffeine
1- Analysis of Illicit narcotics
Nicotine standard was also used as control
A- TLC screening
Test No
Mobile phase Illicit narcotics
Reference standards
4 Toluene: acetone: ethanol: conc. Ammonia(45:45:7:3).
opium alkaloids and heroin.
Codeine
5 Chloroform : Dioxane : Ethyl acetate : Ammonia (29%) (25:60:10:5).
cocaine.
Lidocaine
1- Analysis of Illicit narcotics
Nicotine standard was also used as control
B- Colour tests
1- Analysis of Illicit narcotics
• Also according to UNODC recommendations
Test Materials Illicit narcotics
Marquis test Formaldehyde + conc. H2SO4 +acetic acid
opium alkaloids, heroin and amphetamines
Scott’s Test (modified Cobalt Thiocyanate Test).
cobalt thiocyanate + acetic acid+ glycerin
cocaine.
(Fischer-Morris test) formic acid + sodium nitrite
methaqualone and diazepam
• Applied to tobacco and to spots observed in TLC
A. Identification
by HPLC
C18 column , T Mobile phase : methanol+ water + acetate buffer solution + acetonitrile; UV wavelength: 262 nm
B- quantification
By UV spectroscopy at 262 nm
2- Analysis of Nicotine
RESULTS
Fig. 1 Standard calibration curve for Rf of reference standards (codeine , ephedrine and lidocaine) in test 1 TLC.
1. Analysis of illicit narcotics; A- TLC screening
1. Analysis of illicit narcotics; A- TLC screening
Fig. Standard calibration curve for Rf of reference standards (codeine and ephedrine ) in test 3 TLC
1. Analysis of illicit narcotics; A- TLC screening
Fig. 3 Standard calibration curve for Rf of reference standards (Lidocaine and caffeine) in test 3 TLC.
1. Analysis of illicit narcotics; A- TLC screening
Table. 7 Correction factors for TLC screening using different mobile phases as mobile
phase
Test No. Mobile phase Tested
material
Experimental
standard
Rf x 100
UNODC
Rf x 100
Correction
factor
(C,f.)
4 Toluene: acetone:
ethanol: conc.
Ammonia
(45:45:7:3)
Codeine
standard
38 40 1.052
5 Chloroform:
Dioxane:
Ethyl acetate:
Ammonia (29%)
(25:60:10:5)
Lidocaine
standard
76 77 1.01
• Correction factor = UNODC Rf/ Experimental Rf
1. Analysis of illicit narcotics; A- TLC screening
1. Analysis of illicit narcotics; A- TLC screening
1. Analysis of illicit narcotics; B- Colour tests
Table 9 Results of colour tests of illicit drugs in buccal tobacco brand
Test Illicit drugs Result Interpretation
Marquis Opium alkaloid
and Heroin
(-ve) No opium alkaloid or heroin is
present
Amphetamines
(-ve)
No amphetamines are present
Mod
ifie
d C
ob
alt
Th
iocy
an
ate
Step 1 Cocaine (-ve) No cocaine , tertaciane or
lidocaineor are present
Step 2
Step 3
Fischer-Morris Methaqualone
And diazepam
(-ve) No methaqualone or diazepam is
present
2. Analysis of Nicotine ; A- Identification
Fig. 4 HPLC chromatogram of nicotine reference standard (above) and buccal tobacco ( below).
5.5 min
5.5 min
5.5 Min.
5.5 Min.
1. Analysis of Nicotine ; B- Quantification
% Nicotine : compared to standard nicotine (19.2 mg) / 1 g tobacco dose
Table 11 Content of nicotine per 1 g dose of tobacco brands
Nicotine Content Practical
concentrati
on (Cp)
µg/ml
Absorbance
(Average ±SD)
Theoreti
cal
Conc.
(Ct)
µg/ml
Test
% of daily
usual
consumption
(384 mg)
Ratio Buccal
tobacco:
cigarette
Nicotine
(mg)
%
Nicotine
50 %
135: 1
176.2 898.958 17.26 0.825 ± 0.052 1.92 (1 g)
sachets of
Buccal
tobacco
brand
0.3 % 1.27 6.615 0.127 0.009 ± 0.001 1.92 (1 g) One
cigarette
of a
popular
brand
Conclusions
& Recommendations
CONCLUSIONS
• Based on results obtained in this study, it could be
concluded that
• 1- There are no illicit drugs found in the buccal
tobacco brand investigated in this study
• 2- The buccal tobacco brand contains nicotine
• 3- The buccal tobacco brand contains high
overdose of nicotine.
• 4- The risks of the buccal tobacco brand is not
due to illicit drugs but due to high nicotine
overdose which can cause “Nesbitt's paradox
syndrome “reducing neuronal activity”
RECOMMENDATIONS • The present study recommends
• 1-Authority in Yemen to prevent importing of
the buccal tobacco brand investigated in this
study unless the producer reduce the amount of
nicotine /sachet to normal global allowance.
• 2-To screen cannabis in the buccal tobacco
brand
• 3- Yemeni health ministry to make educational
campaigns about the risks of that buccal
tobacco and directed these campaigns to young
adults, parents and whole family.
THANK YOU