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