DETERMINATION OF ISOPROTURON

BY
UV-VISIBLE SPECTROPHOTOMETER

A.K. Jaiswal1*,Avni Jain2

ABSTRACT

Isoproturon is a carbamate insecticide, which is generally used to control a variety of

insects by both professional pest control workers and homeowners. So far, Gas
Chromatography or High Performance Liquid Chromatography detects it. These methods
are not only costly but time consuming also. Now an attempt has been made to develop a
new method for the analysis of isoproturon in biological samples using UV-Visible
spectrophotometer technique, which is inexpensive, accurate and non-destructive.
Isoproturon was extracted from blood using solvent extraction methods and then
identified by UV-Visible spectrophotometer. The method developed is simple,
inexpensive, accurate and non-destructive that allows for sensitive and reproducible
analysis f isoproturon in various bilological fluids.

KEYWORDS: Isoproturon, UV-Visible Spectrophotometer, cuvettes,Isopropanol.

1. Chemist department of forensic medicine & toxicology, All India Institute Of

Medical Sciences,Ansari Nagar, New Delhi-110029 , tel
9868115165,akjtox1@yahoo.com
2. Student B.tech Biotechnology, Amity Institute of Biotechnology
Noida,avni.j.06@gmail.com

* corresponding author

1
INTRODUCTION

Isoproturon is a broad-spectrum organophosphate insecticide, which is used to control a

variety of insects. Isoproturon is a man-made chemical used as a herbicide. Isoproturon
is a herbicide that can be harmful to wildlife. Isoproturon is released to the environment
during its application as an agricultural herbicide.

Scientifically it is N, N-dimethyl-N?-[4-(1-methylethyl) phenyl] urea.

Structure of Isoproturon

Empirical formulae of isoproturon is C12H18N20 and its molecular weight is 206.3. Pure
isoproturon occurs as colourless crystals, melting a 158 deg C. It is slightly soluble in
water and polar organic solvents, much less so in hydrocarbons. It is stable to light and in
mildly acid and alkaline solution, but hydrolyses in strong alkali and when heated.

The review of literature on analytical methods for isoproturon reveals that the Gas Liquid
Chromatography or High Performance Liquid Chromatography has been extensively
used for the analysis of isoproturon. These methods are not only costly but time
consuming also. An attempt has been made to analyse isoproturon in blood, which is very
cheap and can be performed in any laboratory with very simple instrument i.e. UV-
Visible spectrophotometer.

Experimental

Materials Required

Chemicals and reagents: Isopropanol, hexane and sodium Sulphate dry purified from
Merck India Private Ltd. Mumbai, Isoproturon Standard

Glassware: beaker, test tubes, pipettes, conical flask, volumetric flask, separating funnel,
etc.

Equipment

Double beam UV-Visible spectrophotometer model UV 5704SS (Make ECIL

Hyderabad) was used for the analysis under the following conditions. Two similar quartz
cuvette of path length 1cm were used in this spectrophotometer.

2
 Double beam UV- visible spectrophotometer UV5704SS

Data of mode: Absorbance

Start Wavelength: 200nm

Stop wavelength: 500nm

D2: On

Beam Mode: double beam

Preparation of standard and calibration solutions

1000-ppm solution (stock solution) was prepared in iso propanol by dissolving suitable
amount of known percentage of isoproturn standard. For calibration, solution of 5 ppm,
10ppm, 15ppm, 20ppm were prepared by diluting stock solution with Isopropanol.

Sample preparation

Spiking of blood: 1 ml of 1000 ml of isoproturon was spiked in 2ml of blood sample and
kept overnight in the incubator.

Extraction from blood

10ml blood + 10 ml of 10% Tungstate solution + 15 ml of 5N Sulphuric acid, shaken for

2 minutes

3
 Filtrated

i) Filtrate Residue

ii) Filtrate Residue

iii) Filtrate Residue

Filtrate i), ii) and iii) are pooled + 20 l of hexane is taken in a separating funnel and
shaken for two minutes.

i) Hexane layer aqueous layer

4
 20ml hexane

ii) Hexane layer aqueous layer

20ml hexane

iii) Hexane layer aqueous layer

Hexane layer i), ii) and iii) are pooled and passed through the sodium Sulphate and
evaporated up to 5 ml.

Recording of spectra of standard and sample

Spectra of standard solution were recorded by taking Isopropanol as reference sample.

From these spectra it is clear that λmax is coming between 208 to 241.0nm . Spectra of
different concentrations of isoproturon were recorded. There is one major peak seen at
241.0 nm. Therefore the caliberation graph was drawn at λmax= 241 nm by using
different caliberation standard solutions such as 5ppm, 10ppm, 15ppm, 20ppm.

Results and discussions

Spectra of different concentrations of isoproturon were recorded as shown in fig. There is

one major peak seen at 241.0 nm. λmax was recorded at 241.0 nm.

Table:1- Different concentration of isoproturon v/s Absorbance

Concentration of isoproturon Absorbance at λmax = 241nm

(in ppm)
5 ppm 0.249
10 ppm 0.556
15 ppm 0.921

5
20 ppm 1.125

A calibration graph was drawn at λmax which is shown in fig.The linear range of

concentration varies from 5-20 ppm. The concentration of a sample of isoproturon can be

measured in the linear range, using calibration graph.

REFERENCE

1. Racke KD. 1993. Environmental fate of chlorpyrifos. Rev Environ Contam Toxicol

131:1-50

2. Rack KD. Coats JR. 1990. Comparative degradation of organophosphorus insecticide

in soil: Specificity of enhanced microbial degradation.J.Food Chem 36:193-199.

3. Rack KD. Coats JR. 1990.,Enhanced Biodegradation of Psticides in the environment.

ACS Symposium series 426. American Chemical Society, Washington,DC. 68-81.

4. Rack KD. Coats JR. Titus Kr.1988. Degradation of chlopyrifos and its hydrolysis

product in soil. J Environ Sci Health B23(6): 527-540.

5. Ock Kyoung Chun, Hee Gon Kang, and Myung Hee Kim, 2003 Multiresidue method

for determination of Pesticids in Korean Domestic Crops by Gas Chromatography/

mass Selective Detection. AOAC Vol. 86 No. 4 pp 823-831.

6. Cristina Yagiie et. al ,2001, Multiresidue determination of organochlorine pesticides

and polychlorinated biphenyls in milk by Gas Chromatography with electron capture

detector after extraction by matrix solid-phase dispersion. AOAC Vol. 84 No. 5 pp

1561-1568.

6
7. Salwa M Dogheim et. al 2001, Monitoring of Pesticde residues in Egyptian fruit and

vegetables during 1996. AOAC Vol. 84 No. 2 pp 519-531.

8. Carlo Bicchi et. al, 2001, Simultaneous determination of six triozolic pesticides in

Apple and Pear Pulps by Liquid Chromatography with Ultraviolet Diode Array

detector, AOAC Vol. 85 No. 5 pp 1543-1550.

9. Ana I. et. al, 2001, Determination of five pestisides in orange by Matrix Solid- phase

dispersion and Liquid chromatography to estimate daily intake of consumers. AOAC

Vol. 84 No. 3 pp 891-900.

10. Antonio Valverde 2000, Chromatography pesticides residue analysis : Introduction

AOAC Vol. 83 No. 3 pp 679.

11. Holgervan Lshaut and Wolfgang Schwack,2000, Selective trace determination of

dithiocarbamate fungicides in fruits and vegetables by reversed-phase ion- pair liquid

chromatography with UV and electrochemical detector, AOAC Vol. 83 No. 3 pp

720-727.

12. C. N. Rao, 1961, Ultravoilet & Visible Spectroscopy, London : Butterwonth.

13. A.I.Vogel, 1978 “A text book of quantitative Inorganic Analysis”, 4 th Edition

(Longman Group Ltd London),

7
 DETERMINATION OF ISOPROTURON

BY

UV- VISIBLE SPECTROPHOTOMETER

ABSTRACT

INTRODUCTION

Fig
1: Structure of Isoproturon

EXPERIMENTAL

Materials

8
Equipment

UV5704SS, double beam UV- spectrophotometer was used for the analysis. Two similar
quartz cuvettes of path length 1cm were used in the analysis.

Fig 2: Double beam UV- visible spectrophotometer UV5704SS

1)High resolution grating with high energy efficiency and minimum stray light.
2)High sensitivity photo detector for the entire spectral range. 3)Automatic rapid
scanning of the entire spectrum. 4)User programmable Multi-Standard calibration.
5)Kinetic studies with real-time graphic display. 6)Automatic introduction of filters
at respective wave lengths to minimize stray light effect. 7)Self diagnostic check
facility. Spectrum scanning 8)Wavelength scanning is just as simple as clicking your
mouse. Full scan range from 190 to ll00nm. Data processing capabilities includes: -
9)Full scan 10)Derivative spectrum 11)Smoothening 12)Arithmetic functions 13)Abs
to %T conversion

9
Experimental Conditions

Data of mode : Absorbance

Start wavelength : 200 nm

Stop wavelength : 600 nm

Beam mode : Double

Preparation of standard and calibration solutions

Sample preparation

RESULTS AND DISCUSSION

ACKNOWLEDGEMENT

REFERENCE

10
Isoproturon

Herbicide

NOMENCLATURE
Common name isoproturon (BSI, E-ISO, (m) F-ISO)
IUPAC name 3-(4-isopropylphenyl)-1,1-dimethylurea; 3-p-cumenyl-1,1-dimethylurea
Chemical Abstracts name N,N-dimethyl-N'-[4-(1-methylethyl)phenyl]urea
Other names IPU; ipuron* CAS RN [34123-59-6] EEC no. 251-835-4 Development
codes Hoe 16410 (Hoechst); AE F016410 (AgrEvo); CGA 18 731 (Ciba-Geigy); 35689
RP; LS 6912999
Isoproturon APPLICATIONS
Biochemistry Photosynthetic electron transport inhibitor at the photosystem II receptor
site. Mode of action Selective systemic herbicide, absorbed by the roots and leaves, with
translocation. Uses Pre- and post-emergence control of annual grasses (Alopecurus
myosuroides, Apera spica-venti, Avena fatua and Poa annua) and many annual broad-
leaved weeds in spring and winter wheat (except durum wheat), spring and winter barley,
winter rye, and triticale, at 1.0-1.5 kg a.i./ha. Phytotoxicity Non-phytotoxic to cereals,
except durum wheats. Formulation types SC; WP. Selected tradenames: 'Alon' (Aventis);
'Arelon' (Aventis); 'Strong' (Aventis); 'Tolkan' (Aventis); 'Cordelia' (Griffin); 'Dhanulon'
(Dhanuka); 'Guideline' (Barclay); 'Hilproturon' (Hindustan); 'Iprofile' (Mirfield);
'Isoguard' (Gharda); 'Narilon' (Nagarjuna Agrichem); 'Panron' (Sanonda); 'Pasport'
(RPG); 'Protugan' (Makhteshim-Agan); 'Turonex' (Agriphar); mixtures: 'Javelin' (+
diflufenican) (Aventis); 'Affinity' (+ carfentrazone-ethyl) (FMC); 'Herbaflex' (+
beflubutamid) (Staehler, Ube).

Isoproturon [N-(4-isopropylphenyl)-N_,N_-dimethylurea] (Fig.

1) is a phenylurea herbicide that is used in Europe mainly for
the control of annual grasses and broad-leaf weeds in winter
cereals

Degradation of isoproturon in soil takes place primarily

11
through microbial processes, although photodegradation of
isoproturon has been demonstrated

Microbial degradation
can lead to complete mineralization of isoproturon, which
can mitigate the release of the herbicide to the environment,
but it may also result in metabolites with possible detrimental
properties.

12