Relationship of Tobacco Weight in Commercial Cigarettes to

Nicotine Levels

Mariany Razali

, Muhammad Taufik

, Evi Dayatul Atiqah

, Rid Wanto

, Anny Sartika Daulay

Desi Ardilla

, Endang Susilawati

and Afniwati

Department of Pharmacy, Universitas Tjut Nyak Dhien, Medan, Indonesia

Department of Chemistry, Universitas Sumatera Utara, Medan, Indonesia

Department of Pharmacy, Universitas Muslim Nusantara Al Washliyah, Medan, Indonesia

Department of Agricultural Technology, Universitas Muhammadiyah Sumatera Utara, Medan, Indonesia

Department of Nursing, Poltekkes Kemenkes, Medan, Indonesia

Keywords: Nicotine, Cigarettes, Electrosynthesis, Paper Chromatography, UV Spectrophotometer.

Abstract: Nicotine is a substance contained in tobacco leaves which is used for the manufacture of cigarette raw

materials. Nicotine will come out of tobacco in the process of smoking or inhaling second hand smoke. In

cigarettes the nicotine content should not exceed 1.5 mg. Nicotine is very dangerous for health and is

addictive. This study aims to determine the effect of tobacco weight on nicotine levels and to analyse the

nicotine contained in commercial cigarettes by applying the paper chromatography method and UV-Vis

spectrophotometry. In this study, an experimental method was used, the experimental method was

developed on electro synthetic coupling maceration with variations in sample weights of 0.3, 0.6, 0.9, 0.12,

and 0.15 grams. The qualitative test results obtained in the Paper Chromatography method were nicotine

positive because the Rf value of the sample was close to the Rf value of the standard solution and was

reddish yellow, using electro synthetic coupling maceration with qualitative tests with Cyanogen bromide,

the best results were marked by abundant greenish yellow colour . Using the UV-Vis spectrophotometry

method, it was obtained that in each increase of 0.3 to 1.5 grams of tobacco, the concentration would

increase by 0.4 ppm. Paper Chromatography Methods and UV-Vis Spectrophotometry can be used to

analyse nicotine and produce the best results.

1 INTRODUCTION

Tobacco leaves contain the alkaloid nicotine, a type

of neurotoxin (Baranska, Kaczor, and Chruszcz-

lipska 2012). This substance is often considered evil,

which makes tobacco products banned. Nicotine will

come out of tobacco in the process of smoking or

inhaling secondhand smoke or chewing (Rahmat

Nur Hidayat, Adam M. Ramadhan 2016). In pristine

leaves, nicotine is bound to organic acids and

remains bound to acids when the leaves are slowly

dried. Tobacco is deadly when consumed 60 mg at a

time. It is the same as when the body receives water

consumption that exceeds the capacity that is usually

tolerated. In the case of cigarettes, the nicotine

absorbed in the body is 10% of the nicotine content

in a cigarette. If the weight of tobacco in a cigarette

is 0.76 g, with 2% nicotine content in tobacco

leaves, then the cigarette contains 15 mg of nicotine

and 10% is absorbed by the body or the equivalent

of 1.5 mg (Solarino et al. 2009).

Nicotine is the main pharmacologically active

component of tobacco, and is also found in large

quantities in other species in the Solanaceae family

(Wiencek et al. 2019). At low concentrations

nicotine is a stimulant, namely nicotine increases

activity, alertness, and memory. This is one of the

factors that contribute to the dependence on tobacco

smoking. Nicotine can increase heart rate, blood

pressure, and reduce appetite. At high doses,

nicotine acts as a depressant or suppressant (Rahmat

Nur Hidayat, Adam M. Ramadhan 2016).

Cigarettes are one of the tobacco products that

are intended to be burned, smoked or inhaled,

including clove cigarettes, white cigarettes, cigars or

other forms produced from the nicotiana tabacum,

nicotiana rustica, and other species or synthetics

whose smoke contains nicotine and tar. , with or

Razali, M., Tauﬁk, M., Atiqah, E., Wanto, R., Daulay, A., Ardilla, D., Susilawati, E. and Afniwati, .

Relationship of Tobacco Weight in Commercial Cigarettes to Nicotine Levels.

DOI: 10.5220/0010614800002775

In Proceedings of the 1st International MIPAnet Conference on Science and Mathematics (IMC-SciMath 2019), pages 621-625

ISBN: 978-989-758-556-2

621

without additives (Fidrianny 2004). Cigarettes are

usually cylindrical of paper with a length of between

70 and 120 mm (varies by country) with a diameter

of about 10 mm which contains chopped tobacco

leaves (Hamdan 2018).

Extraction is the process of separating

materials from the mixture using a suitable solvent

(Gupta and Kothari 2014). The extraction process is

stopped when an equilibrium is reached between the

concentration of the compound in the solvent and

the concentration in plant cells. After the extraction

process, the solvent is separated from the sample by

filtration (Rusevska and Zdravkovski 2011). The

initial extract is difficult to separate through a single

separation technique to isolate a single compound.

Therefore, the initial extract needs to be separated

into fractions that have the same polarity and

molecular size (Jahed, Hamidi, and Galehassadi

2020).

Nicotine extraction from cigarettes can be done

in several ways, namely maceration extraction,

reflux, and distillation. Extraction by maceration

provides the advantage of undamaged samples and

more yields (Rusevska and Zdravkovski 2011).

Maceration is a simple method of extraction.

Maceration is done by soaking the simplicia powder

in a liquid filter. The fluid will penetrate the cell

wall and enter the cell cavity containing the active

substance, the active substance will dissolve because

of the difference in concentration between the active

substance solution in the cell and the one outside the

cell, the concentrated solution is pushed out. These

events are repeated so that there is a balance of

concentration between the solution outside the cell

and inside the cell (M Taufik et al. 2018).

The extraction of nicotine from cigarettes can

be done by means of electrosynthetic maceration

because it is a way to synthesize or produce a

substance which is based on electrochemical

techniques (Muhammad Taufik et al. 2021). In this

method, there is a change in the element or chemical

compound into the desired compound (Taufik,

2016).

Hidayat (2016) has analyzed nicotine using the

ultraviolet spectrophotometric method, where the

extraction of cigarettes is carried out with methanol

which is then stirred for 30 minutes. Then add 2M of

NaOH and aquadest then stir again for 5-10 minutes

on the hotplate with a temperature of 700C. Add

zinc acetate and potassium ferro cyanide before

being centrifuged for 10 minutes at a speed of 3000

rpm. The supernatant obtained is added with

petroleum ether and separated using a separating

funnel and the fraction of petroleum ether is

extracted. The determination of the nicotine content

of cigarette samples was carried out by ultraviolet

spectrophotometry at a wavelength of 262 nm with

different concentrations of 20,40,60,80,100, 120 and

the absorbance of 0, 0.5, 1, 1.5, 2, 2.5 in order to

obtain the equation regression y = 0.019 + 0.067 at a

price of R2 = 0.983 (Rahmat Nur Hidayat, Adam

M. Ramadhan 2016).

Hidayat (2015) has reported that accurate

information is obtained regarding the nicotine

content of herbal cigarettes, a research on the

identification of secondary metabolites and analysis

of nicotine levels in herbal cigarettes was carried out

using the UV-Vis spectrophotometer method at a

wavelength of 262 nm. The materials used in this

study included samples of herbal and conventional

cigarettes, methanol, aquadest, petroleum ether, 2 M

NaOH, zinc acetate, and potassium. Cigarette

extraction was carried out with methanol plus 2 M

NaOH and aquadest on a hotplate with a temperature

of 70°C. Zinc acetate and potassium ferrocyanide

were added before being centrifuged for 10 minutes

at 3000 rpm. The supernatant obtained was added

with petroleum ether and separated using a

separating funnel and the petroleum ether fraction

was taken. The results obtained by herbal cigarettes

have a higher nicotine content than conventional

cigarettes. Even though the nicotine levels listed on

the herbal cigarette packaging are very low, even

close to 0 (Hidayat, Siradj, and Selatan 2015)

Commercial cigarettes are cigarettes that are

sold in the market. This cigarette product is a

cigarette product that is in great demand by the

public. Due to advertising promotions that do not

explicitly invite or persuade someone to smoke.

Thus increasing the target consumer to have

smoking behavior. The tendency of society to

understand that commercial cigarettes are no more

dangerous. Actually, commercial cigarettes are the

same as other cigarettes, only differentiating the way

of promoting these cigarettes (Muhammad Taufik et

al. 2017).

The effect of nicotine levels in tobacco found in

cigarettes greatly affects health, so it must be known

the levels contained in each gram of tobacco weight

in cigarettes. This study aims to determine the effect

of tobacco weight on commercial cigarettes on the

levels of nicotine produced, to analyze the

qualitative nicotine in commercial cigarettes using

the Paper Chromatography method, and to analyze

the nicotine contained in cigarettes using the UV-Vis

Spectrophotometric method.

IMC-SciMath 2019 - The International MIPAnet Conference on Science and Mathematics (IMC-SciMath)

622

2 METHODS

Analysis of nicotine in cigarettes using experimental

methods. The experimental method was carried out

using electro synthesis with a time of 15 minutes and

using variations in sample weight successively 0.3 g,

0.6 g, 0.9 g, 0.12 g and 0.15 g. Samples were

analyzed qualitatively using Cyanogen bromide until

a greenish yellow color was obtained and tested

using the paper chromatography method. The

quantitative test was carried out using the UV

spectrophotometer method.

2.1 Materials

The materials used in this study were filter

cigarettes, reagent pH, methanol, chloroform,

aquadest, dragendroff reagent, aluminum foil, filter

paper, and paper chromatography. The tools used in

this research are glass tools in the laboratory such as

measuring cups, beaker glass, test tubes, petri dishes,

Erlenmeyer, a set of electro synthetic tools, a set of

paper chromatography tools and a UV-Vis

spectrophotometer.

2.2 Collecting Sample

Samples were collected used purposive sampling

method, it was carried out deliberately by directly

selecting researchers who met the sample criteria.

The part of the cigarette that is taken is the cigarette

part of the filter

2.3 Preparation

2.5 grams of nicotine standard solution is put into a

1000 ml measuring flask. Gradually dissolve with

methanol while shaking, until it reaches the marking

line. From this solution, a dilution is made with a

concentration of 0.5 ppm, 1 ppm, 1.5 ppm, 2 ppm

and 2.5 ppm.

2.4 Extraction

Filter cigarettes as samples were weighed with

successive variations of 0.3 g, 0.6 g, 0.9 g, 1.2 g, and

1.5 g, put the sample into a 100 ml glass beaker plus

methanol solvent macerated electro synthesis with a

time of 15 minute. The results of electro synthetic

coupling maceration were filtered with filter paper

and then put into a petri dish, allowed to evaporate

until the nicotine was obtained. The nicotine

obtained was dissolved with 20 ml of methanol and

then measured the pH (pH 9).

2.5 Paper Chromatography

Chromatography paper is made horizontal lines on

the bottom edge of 2 cm and the top edge of 3 cm.

Creepage distance is 10 cm. The standard solution

(nicotine) and the test solution were spotted on the

chromatography paper that had been activated

beforehand. The marking is carried out on a

horizontal line on the bottom edge of the paper

chromatography with a spacing of 2 cm. The volume

of the solution is 5 µl, the diameter of the dots

should not be more than 0.5 cm. Then insert the

chromatography paper into the chamber, and closed.

Mobile phase: methanol: chloroform (50: 50). Left

and observed until the spot rises and the solvent

surface reaches the 10 cm limit mark. Then remove

the chromatography paper plate and dry it. Observed

the appearance of spots on paper chromatography

using dragendroff reagent. The colored spots give a

reddish yellow color, then the spots are marked from

the standard solution and the test solution. The Rf

value is calculated based on existing data.

2.6 UV Spectroscopy Analysis

The sample of filter cigarettes as a result of electro

synthetic coupling maceration was analyzed

quantitatively using UV-Vis spectrophotometry.

Where the working conditions in spectrophotometric

analysis are as follows:

1. The cuvette used was a glass cuvette with a

thickness of 10 mm in a square shape.

3. The wavelength used in nicotine analysis is 262

nm.

4. The cuvette containing methanol is inserted into

the spectrophotometer and press the blank button

with a wavelength of 262 nm.

5. The cuvette containing methanol was replaced

with a sample of the nicotine solution from the

maceration results.

6. Wait for the absorbance reading on the

spectrophotometer to stop and show a fixed

number.

3 RESULTS

3.1 Preparation

The standard solution used is a solution containing

the precisely known concentration of the element.

The process of making standard solutions is carried

out by diluting with five concentrations, namely 0.5,

1.0, 1.5, 2.0, and 2.5 ppm.

Relationship of Tobacco Weight in Commercial Cigarettes to Nicotine Levels

623

3.2 Preparation

Smoker's saliva preparation was carried out in the

laboratory of the University of North Sumatra,

Medan. The samples used were 10 ml of active

smoker's saliva with the addition of chloroform

solvent, in the smoker's saliva there is a nicotine

compound that comes from cigarette consumption,

which is directly exposed to cigarettes and cigarette

smoke through the mouth where saliva is contained.

Smoker's saliva has 2 layers perfectly where the

bottom layer of nicotine and the top layer of the

saliva remains. Non-smoker's saliva does not have a

perfect 2-layer separation so it takes 5 minutes to let

the saliva and solvent split into two layers.

3.3 Extraction

The extraction process by means of electro synthetic

coupling maceration has a very important role in the

compounds contained in the filter cigarette sample.

Optimization is carried out through continuous

observation of the aspects that affect the inclusion of

active compounds in the sample to be analyzed. In

this method, the electro synthetic coupling

maceration takes 15 minutes for the maceration

process.

In this electro synthetic coupling maceration

extraction method, a chemical element or compound

changes into the desired compound. The use of

methods in synthesizing materials is based on the

various advantages offered such as the equipment

required is very simple, which consists of two or

three electrode rods connected to an electric current

source.

3.4 Paper Chromatography

Paper chromatography is an analytical method used

to separate colored chemicals, especially pigments.

Apart from being easy and cheap, chromatography

also has the advantage that the resulting spots appear

directly on the chromatography paper. Spraying is

done using dragendroff so that the spots are clearer.

In this study, the results of the paper

chromatography test were used to identify the Rf

value of the test sample, where the sample was said

to be positive for nicotine, if the Rf value of the

sample was the same or close to and the Rf value of

the standard solution. In addition, it is said to be

positive for nicotine when the spots are reddish

yellow. This can be seen in the following Figure

3.1. :

Figure 3.1: The results of the analysis used paper

chromatography.

Figure 3.1 shows that the Rf value in the standard

solution is 0.84 and in samples with 5 variations in

sample weight, namely the sample 0.3 with an Rf

value of 0.76, for a sample 0.6 with an Rf value of

0.77, for a sample 0.9 for an Rf value of 0.8 , sample

1.2 Rf value 0.81, sample 1.5 Rf value 0.83 and the

average Rf value obtained is 0.80. From these data it

can be said to be positive for nicotine because the Rf

value of the standard solution is close to the Rf value

of the sample solution. In addition, the resulting spot

color is orange.

3.5 UV Spectroscopy Analysis

Tthe wavelength at a concentration of 2 ppm and an

absorbance of 1.046 with a wavelength of 262 nm.

In this study, the calculation of the concentration

of quantitative analysis by UV-Vis spectro-

photometry was carried out using the regression

method, namely by using a regression equation based

on the standard concentration of nicotine and the

absorption rate. The results of the regression equation

can be seen in Figure 3.2. the following:

Figure 3.2: The regression equation in nicotine cigarettes

analysis.

Figure 3.2. shows that the regression equation for

filter cigarettes and non-filter cigarettes is Y =

2.568x - 0.767 and R2 = 0.9141. The regression

equation can be used to calculate the nicotine

concentration in filter cigarettes. The results of the

sample concentration can be seen in Figure 3.3 the

following:

0,72

0,74

0,76

0,78

0,8

0,82

0,84

0,86

LB 0,3 0,6 0,9 0,12 0,15

RfValue

comparisonsample

Sample

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Figure 3.3. The concentration of the filter cigarette

sample.

Figure 3.3. shows that the heavier the sample the

heavier the nicotine concentration and each gram of

the sample has a different nicotine concentration.

Therefore, every tobacco that is in a cigarette has a

different nicotine content. Nicotine levels in

cigarettes have a very negative impact on health and

can become addictive. From these data shows the

highest yield of nicotine at a sample weight of 0.15

g, obtained by the micro synthetic coupling

maceration method with a concentration of 0.7 ppm.

However, in this study, we obtained a model on a

lab scale that in each increase of 0.3 to 1.5 grams of

tobacco, the concentration will increase by 0.4 ppm.

4 CONCLUSION

Preparation and extraction of nicotine in cigarettes

using the electro synthetic coupling maceration

method with time variations and methanol solvent

produced nicotine compounds with a positive

greenish yellow color in the qualitative test of

cyanogen bromide. The Paper Chromatography

method can be used to analyze nicotine with positive

results for the presence of nicotine because the Rf

value of the standard solution is close to the Rf value

of the sample besides the resulting spot color is

orange. However, UV spectroscopy can be used to

analyze nicotine with the result that each increase of

0.3-0.15g of tobacco yields a concentration of 0.4

ppm using the Y = 2.568x - 0.767 line.

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