Extraction and Analysis of Nicotine in the Urine of Active Smokers

after given by Vitamin C

Endang Susilawati

, Muhammad Taufik

, Afniwati

, Mariany Razali

, Desi Ardilla

, Rid Wanto

and Virda Lola Br Tarigan

Department of Nursing, Poltekkes Kemenkes, Medan, Indonesia

Department of Chemistry, Faculty of Mathematics and Natural Sciences, Universitas Sumatera Utara, Medan, Indonesia

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

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

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

Keywords: Nicotine, Urine, Active Smoking, Extraction, UV-Vis spectrophotometry

Abstract: Nicotine is a class of addictive compounds that can cause dependence. Vitamin C contains antioxidants that

can ward off free radicals in the body. Nicotine extraction and analysis are a series of processes that are

indispensable in order to produce nicotine in optimal concentrations. This study aims to extract and analyze

nicotine in the urine of active smokers after giving vitamin C. The descriptive method was developed for the

extraction of nicotine in urine. Preliminary test using Cyanogen bromide showed the presence of nicotine in

the urine of smokers for 7 days. The same thing is evidenced by the thin layer chromatography data

resulting in an average Rf value of 0.6. The concentration of nicotine in urine decreased from the first day of

vitamin C administration to day 7. This indicates that there is an effect of vitamin C on the urine of active

smokers.

1 INTRODUCTION

Cigarettes are one of the most dangerous deadly

objects in the world (Rahmat Nur Hidayat, Adam M.

Ramadhan 2016). Consuming cigarettes

continuously can cause various health problems such

as cancer, respiratory problems, and heart problems.

Cigarettes are one of the biggest contributors to

death today. The death rate caused by smoking in

Indonesia has reached 57,000 people every year

(Sari, Zulaikhah, and PH 2019).

Cigarette consumption in Indonesia reaches 215

billion sticks per year. In Indonesia there are 60%

smokers, 59% of whom are men and 37% are

women. In Indonesia, tobacco plus cloves and other

ingredients are mixed to make kretek cigarettes. In

addition to kretek, tobacco can also be used as rolled

cigarettes, white cigarettes, cigars, pipe cigarettes,

and smokeless tobacco, cylinders of paper

measuring 70-120 mm in length and 10 mm in

diameter containing chopped tobacco leaves. The

basic ingredient of cigarettes is tobacco.

Tobacco contains a variety of chemicals that can

be addictive to a person, even if they don't want to

try again (Solarino et al. 2009).These health

problems are related to the content of free radicals in

cigarette smoke. in one suction, an estimated 1014

free radical molecules enter the body (Rahmat Nur

Hidayat, Adam M. Ramadhan 2016). Cigarette

smoke contains ± 4000 chemical compounds with 60

chemical compounds that have been identified as

cancer-causing and genotoxic. These chemical

compounds circulating in the testicular blood vessels

damage the spermatozoa with their cytotoxic effect.

These chemical compounds consist of nicotine, tar,

carbon monoxide, acetone, arsenic, ammonia,

hydrogen cyanide and so on (Clayton et al. 2013).

Vitamin C is one of the most widely taken

nutritional supplements (Pacier and Martirosyan

2015). Health professionals as well as a number of

health benefits for vitamin c such as boosting

immunity or preventing common diseases and

cancer (Lorensia et al. 2018). If free radicals are too

high, natural anti-oxidants alone are not enough to

ward off free radicals. Therefore, additional

antioxidants are needed to inhibit oxidation

Susilawati, E., Tauﬁk, M., Afniwati, ., Razali, M., Ardilla, D., Wanto, R. and Tarigan, V.

Extraction and Analysis of Nicotine in the Urine of Active Smokers after given by Vitamin C.

DOI: 10.5220/0010614400002775

In Proceedings of the 1st International MIPAnet Conference on Science and Mathematics (IMC-SciMath 2019), pages 601-604

ISBN: 978-989-758-556-2

601

reactions, neutralize and scavenge free radicals.

Water-soluble vitamins that can function well as

antioxidants are commonly known as vitamin C or

ascorbic acid. This vitamin C plays an important role

in protecting cell damage caused by free radicals.

Because structural vitamin C is similar to glucose

and can replace glucose in various chemical

reactions (Pacier and Martirosyan 2015).

Nicotine extraction from smoker's urine can be

done in several ways, namely extraction, maceration,

reflux, and distillation. Extraction by means of

maceration gives an advantage, so that it is not

damaged and so that more tends to be obtained.

Extraction by maceration was identified by

sonication and electrosynthesis methods (Taufik et

al. 2017). Fidrianny (2004) has carried out the

preparation and analysis of nicotine contained in

smoker's urine where chloroform is used as a solvent

in sample preparation The electrolysis coupling

maceration method is the best extraction method

compared to the results of the maceration method

(Muhammad Taufik, Rid Wanto, Athaillah, Anny

Sartika Daulay, Lilis Karlina Siahaan, Desi Ardilla,

Mariany Razali 2017). However, this study has a

weakness where the solvent used has not been

optimized. In this work, the use of UV spectroscopy

was also developed at a wavelength of 260 nm.

However, this study is a basic study in order to

search for nicotine in the urine of smokers simply in

the laboratory.

2 MATERIALS AND METHOD

2.1 Sample Collection

The sample was collected purposively, namely based

on the criteria determined by the researcher. The

sample obtained was the urine of volunteers who

consumed cigarettes that had been given vitamin C

and not given vitamin C. The samples were taken

within 24 hours after the volunteers consumed

cigarettes. The urine sample used was from Garu II

Medan Amplas. The dose of with Vitamin C = 50

mg x 2. Samples were taken until day 7.

2.2 Administration of Vitamin C

Filling in the questionnaire sheet including name,

age. Followed by giving vitamin C 50 mg / tab x 2

for 2 times a day morning and evening to volunteers

for 7 days.

2.3 Preparation and Extraction

Each 25 ml active smoker's urine sample was added

with 25 ml of chloroform inserted into a separating

funnel. The lower layer was put into a 50 ml beaker

glass and followed by maceration of the

electrosynthetic coupling for 15 minutes. The result

of maceration is evaporated until the solvent

evaporates. The nicotine evaporated was diluted at

pH 9 then continued with qualitative and

quantitative analysis.

2.4 Preliminary Test using Cyanogen

Bromide

The extracted sample was dripped with Cyanogen

bromide reagent until a yellow color was obtained

which indicated positive nicotine, observed the

yellow color that occurred and compared it with the

standard differentiated into + (slightly), ++

(moderate), +++ (abundant).

2.5 TLC Analysis

Thin layer chromatography (TLC) analysis was

performed using the mobile phase of Methanol:

Chloroform (50:50). The stain appearance solution

used was dragendorff reagent. Physically, the stain

analysis uses UV light.

2.6 UV Analysis

The results of the smoker's urine sample which was

macerated with electrosynthetic coupling were

analyzed quantitatively using MDA 50 UV-Vis

spectrophotometry. This instrument was switched on

for 10-15 minutes and used a wavelength of 200-800

nm. The cuvette used was a glass cuvette with a

thickness of 10 mm, a square shape. In this work, the

blank used is methanol.

3 RESULTS AND DISCUSSION

3.1

Preparation and Extraction

The sample was taken from 5 respondents who were

active smokers. The preparation has a very

important role in determining the success of the

extraction and analysis process. The extraction

process uses chloroform as a solvent. This sample

extraction aims to separate nicotine from the urine of

active smokers. The electro synthetic method was

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

602

developed to shorten the extraction time. This

technique is a way to synthesize which is based on

electrochemical techniques. In this study, the electro

synthetic coupling maceration was developed for 15

minutes with a voltage of 20 V.

3.2

Preliminary

Test using Cyanogen

Bromide

Preliminary test results using Cyanogen bromide can

be seen in Table 1 in the following:

Table 1: Preliminary test results

Sample Da

1 2 3 4 5 6 7

A ++ ++ ++ + + + +

B + ++ ++ ++ + + +

C ++ ++ ++ + + + +

D ++ ++ ++ + + + +

E ++ ++ + + + + +

Table 1 showed that the urine of active smokers

in the analysis showed positive results for nicotine

compounds. This is indicated by the appearance of

orange deposits. In this study, the greatest nicotine

was obtained on the third day then decreased until

the 7th day.

3.3 TLC

Analysis

The results of TLC analysis is presented in Table 2

as follows:

Table 2: Sample Rf value

Sample Day Stain

distance

I 6.5 0.92

II 6.6 0.94

III 6.6 0.94

IV 6.5 0.92

V 6.6 0.94

VI 6.6 0.94

VII 6.6 0.94

VIII 6.6 0.94

I 6.4 0.91

II 6.3 0.90

III 6.2 0.88

IV 6.0 0.85

V 5.9 0.84

VI 6.0 0.94

VII 6.0 0.94

I 6.6 0.94

II 6.6 0.94

III 6.6 0.94

IV 6.6 0.94

V 6.6 0.94

VI 6.6 0.94

VII 6.6 0.94

VIII 6.1 0.87

I 6.1 0.87

II 6.2 0.88

III 6.6 0.94

IV 6.6 0.94

V 6.6 0.94

VI 6.6 0.94

VII 6.6 0.94

VIII 6.6 0.94

I 6.6 0.94

II 6.6 0.94

III 6.6 0.94

IV 6.7 0.95

V 6.5 0.92

VI 6.5 0.92

VII 6.6 0.94

VIII 6.6 0.94

icotine

standar

6.6 0.94

Table 2 shows that the Rf values of samples A,

B, C, D, and E from Day 1 to 7 are almost the same

as for standard nicotine (Rf = 6.6).

3.4

Quantitative

Analysis using UV

UV-Vis spectrophotometry was developed to

generate quantitative data. Initially, the standard

solution was made from nicotine standard solutions

with concentrations of 0.5, 1.0, 1.5, 2.0, and 2.5

ppm. The blank used is methanol. To determine the

nicotine concentration, first the maximum

wavelength must be determined at 260 nm.

Determination of wavelength used a concentration

of 2 ppm. The results of the analysis of nicotine

concentration for the 5 samples used can be seen in

Figure 1.

Figure 1: The results of the analysis of nicotine levels

Figure 1 showed data from five urine samples of

active smokers that had been extracted using

Extraction and Analysis of Nicotine in the Urine of Active Smokers after given by Vitamin C

603

chloroform. The nicotine concentration table in the

sample can be seen in Table 3.

Table 3: Nicotine concentration

Figure 1 and Table 3 shows the nicotine

concentration generated from the five analyzed

samples. The concentration of urine generated from

day 1 to day seven is reduced. This shows that

Vitamin C given to active smokers has an effect on

urine levels.

4 CONCLUSION

The extraction of nicotine in the urine of active

smokers can be developed using electro synthetic

coupling maceration. The extraction results were

analyzed using a preliminary test and thin layer

chromatography indicated the presence of nicotine

in the smoker's urine. The urine concentration

decreased until the 7th day after administration of

Vitamin C by testing using UV spectroscopy at a

wavelength of 260 nm.

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