Mutagenicity Assessment of Textile Dyes by Using Ames Test

Project Report, 2017

38 Pages, Grade: master












I would first like to thanks my thesis advisor Dr. Joginder Singh Panwar, Associate Professor, School of Biosciences, Lovely Professional University for his wise and painstaking guidance that he has given throughout this research work. His generous co-operation, suggestions, Constant encouragement and kind supervision, made this dissertation possible.

I express my gratitude to Dr. Neeta Raj Sharma, Professor and Head of School of Biosciences, Lovely Professional University their kind support and encouragement for my Research endeavor. I am highly indebted to Deepika Ma ’am for her frequent help, care and Encouragement during my study period of completing this thesis. I would also like to express my deep thanks to my friends especially (Ayan Das) whose selfless cooperation helped me in my thesis. I am also thankful to staff members of library and all other teaching and non-teaching staff especially (John Masih) senior techncian of Lovely School of Biosciences, Lovely Professional University, for their kind care and cooperation.

Finally, I respectfully offer thanks to my beloved parents, especially my mom (Kamaljit kaur) For their love, mellifluous affection and sincerity which hearten me to achieve success in every Sphere of my life. I also pay thanks to my beloved sister (Jaspreet kaur), and to my husband (Kirandeep Singh) regarding their love, moral support and constant encouragement. Without their Encouragement and support, the present study would have been a mere dream.


Chapter: 1


“The earth provides enough to satisfy every person’s need, but not every person’s greed”

-Mahatma Gandhi

The human life fitting to the natural variety of the earth’s system and climate. The earth consists of plants, animals, human being, air, water, soil and land. Some conditions are necessary for the development of our life on earth. If these condition are present in unlimited from then it cause some problem ‘As we know excess of everything is bad’. Activities done by human into natural environment are so widespread and silence in their consequences that they too affect the earth.

The researches done by scientists are also paly important role in protecting theEnvironment. Researchers tell the advantages of environment to the humans health economic, social and aesthetic harm. Result for this whole is that we are now aware that how we protect our environment from these natural hazardous things.

The environment which is free from these contaminations i.e. the environment which is safe and hygienic should provide us clean water, soil, and natural environment. In the past, men rely on inexhaustible resources which are provided by environment to us without a limit and the law of nature directed our evolution. But now in present world we started supplements on muscle power with exhaustible resources, coal, oil, and uranium. As a result, in some aspect We are directed our own evolution.

Environment pollution

Nature is a gift of God. We have no any right to destroy its beauty. We should not destroy its beauty and if we do so, we will have to face the consequences. We should take care of nature, because it is a basic need of our life but now pollution is the major problem on earth. Pollution is defined as the introduction of contaminants into the environment that results adverse change in environment. It is one of the major issues in the modern world. Pollution can be present in environment in the form of chemical substances or energy such as water, heat and light. Many Countries in the world having a specific connection with the effects of contaminated drinking water because of that water-borne diseases are a more common or increasing in the environment that lead to the major cause of morbidity and mortality (Clasen et al. 2007; WHO 2010). Contamination free drinking water is great significance for overall health and have considerable importance in young and infant child health and their survival (Anderson et al. 2002; Fewtrell et al. 2005; Ross et al. 1988; Vidysagar 2007). The World Health Organization (2005) estimates that worldwide about 1.8 million people die from diarrheal diseases. Persons with weak immune systems, such as those peoples who was suffering from AIDS are especially harmed to water- borne infections, even those peoples which are not typicallys howing an intention to cause bodily harm to healthy individuals (Kgalushi et al. 2004; Laurent, 2005)

Types of pollution Air pollution

Release of chemicals into natural environment cause air pollution. For example chemicals like carbon monoxide, chlorofluorocarbon, and nitrogen oxide which is produce by industry and motor vehicles into the natural environment.

Light pollution

In light pollution light trespass, over illumination and astronomical interference is a major cause of pollution into the environment.

Noise pollution

Noise pollution includes aircraft noise, industrial noise, roadways motor vehicles noise which makes peoples sick and tensed. Also due to noise people brain not able to take rest.

Soil pollution

Soil pollution occur when soil are exposed to heavy metals, hydrocarbon, M TBE herbicides, pesticides, or chlorinated hydrocarbons.

Water pollution

Contamination which occurs in water bodies such as lakes, rivers, underground water etc. is known as water pollution. In this we mainly consider the contamination which is occur due to textile dyes. It occurs directly and indirectly into the water.

Pollution caused by textile industries

Dyes are mainly used to produce consumer’s products like paints, textile printing inks, paper, and plastics. They are very complex and sensitive chemicals. They add color and pattern to the material. Many different vat dyes and dispersed dyes were used in the textile industries.

Textile dyes industries causes wide range of pollution in the world. The World Bank roughly calculated about 20% of the global industrial water pollution originates from the treatment and dyeing process in the textile industries. Textile production of dyes predicted to release aromatic amines (Benzedrine and toluidine), ammonia, alkali salts, toxic solids and large amount of pigments, chlorine, a known carcinogen. The dyes which are not treated cause chemical and biological changes in our aquatic system and human life. The presences of this compound also make water unhealthy and dangerous to human life. The polluted water also affects the soil health and agriculture field by contaminating its effect the germination and growth of crops. The textile industry in Ludhiana city in Punjab produces high water pollution which is dangerous to peoples in the region. Textile Dyes Synthetic dyes are majorly used in textile industries for dyeing, paper printing, color photography, pharmaceutical, food, cosmetics and other industries (Rafi, Franklin and Cerniglia,1990). It was estimated that about, 10,000 different dyes and pigments are used industrially, and over 7x105tons of synthetic dyes are produced every year worldwide. In 1991, the world production of dyes was find approximately about 6, 68, 000 tones out of which azo dyes contribute about 70 % (ETAD, 1997). During dying process huge amount of azo dye is lost in wastewater.

Types of dyes

The first invented man made organic dye was, maven, which is discovered by scientist William Henry Perkin in 1856. Thousands of synthetic dyes have since been prepared, synthetic dyes can being replaced by natural dyes. Classification of dyes given below:

1. Acidic dye
These are the dyes which are applicable to the fibers such as silk, wool, and these acidic dyes are soluble in water.
2. Basic dyes
These are mainly applicable to a acrylic fibers for wool and silk. These dyes are water soluble cationic dyes. These dyes are also used for the coloration of paper.
3. Direct or substantive dyes
By the addition of sodium chloride or sodium sulphate direct dyes are normally carried out at slightly or neutral dye bath. These dyes are used on cotton, paper, leather, wool, silk and nylon etc. These dyes are also used as PH indicators.
4. Mordant dyes
The most important mordant dyes are synthetic mordant dyes or chrome dyes. They applied to wool. These mordant dyes contain heavy metals that are dangerous to health and extreme care has been taken when we use them.
5. Vat dyes
Vat dyes are water insoluble and not able to dying the fiber directly. However, alkaline liquor reduction produces a water soluble alkali metal salt of the dye.
6. Reactive dyes
These are the dyes which utilized the attached chromosphere to substituents that are capable of directly reacting with fibers substrate. The colvant bonds that attaches the reactive dye to normal fibers to make them most permanent dyes..
7. Dispersed dyes
Dispersed dye is water insoluble dye and also used to dyeing cellulose acetate and they can also use to dye nylon, cellulose, triestate and acrylic fibers.
8. Other important dyes
There are number of other classes of dyes including oxidation bases, laser dyes, and leather dyes, solvent dyes, carbine dyes, carbine dyes.

The chemicals which are used to produce dyes today are highly carcinogenic, or even explosive. Aniline is the popular group of dyes known as azo dyes which are extremely poisons and dangerous to handle and work with, also being highly burnable. Other harmful chemicals which are used in the dying process include:

1. Dioxin – a carcinogen and possible hormone disrupter;
2. Toxic heavy metals such as chrome, copper and zinc-known carcinogens; and
3. Formaldehyde, a suspected carcinogen.

Disperse dye almost totally insoluble in water it exists as a dispersion of microscopic particles, with only small amount in true solution. Disperse dye extensively used in textile industries for dyeing. Azo dyes which are widely used in textile industries which are one of the largest class of synthetic organic dyes and that having nitrogen as the azo group. Azo dye has been shown to destroy natural ecosystem when these azo dyes was discharged into water system. At the same time, the more drastic dyeing conditions and improved methods which developed along with the newer fibers have allowed the use of more complex molecular structures, although many acetate dyes are still used on polyesters and nylon. Azo, anthraquinone, o-nitro diphenylamine, methine and quinophthalone chemical classes provide the major commercial disperse dyes. None of these is a new chromophoric system, but intensive investigation and novel elegant synthetic methods have resulted in dyes exhibiting superior properties, especially on the newer fibers. Disperse dyes are also used in the newly popular transfer print process, which requires coloring matters of poor sublimation fastness, a source of trouble in conventional printing.

The accurate amount of dye produced in the world is not known. Exact data on the quantity of dyes discharge in the environment are also not available. Globally, accessing to freshwater is becoming more acute every day. Studies have shown that many microorganism catabolise these synthetic dyes as sole source of carbon and energy. Instead of dispersed or azo dyes there are also many other dyes which causes water contamination. These all dyes contaminants the water and through water it can enter into human body and cause mutation which results into various effect to human health it can also be carcinogenic that is cause cancer in human.

.Studies have also shown that many microorganisms catabolise these synthetic dyes as sole source of carbon and energy. It was shown that some bacteria can also be used to degrade complex Azo compounds like 1-(4’-carbophenylazo)-4-naphthol. Hence, various type of dyes which are use at large scale in industries containing carcinogenic substances which when thrown into water cause contamination of water that lead to cause various disease in human. Different dyes which we use in textile industries are:

Hence, dyes which used in textile industries are more harmful for the people who live near the area. Microorganism growing in the industries effluents containing various dyes are cultured and studied for their potential in degrading those harmful dyes. These microbes basically use these dyes as a source of carbon, nitrogen, energy



The research work aim was to assess the mutagenicity or carcinogenicity of textile dyes to the people live near to the textile mills.


- Survey of Ludhiana textile mills for knowing different disperse and vat dyes used throughout the year
- To perform an assay to determine the mutagenicity of disperse and vat dyes (Ames test)
- Data Analysis for Ames test


Textile dyes were used in colouring the clothes.Textile wastewater includes various types of dyesand chemicals that make the environmental challenge for textile industry. Main pollutants in textile wastewater came from dyes.

Textile industry effluent contains these toxic dyes. These toxic compounds enter into the surface water contaminate the surface water and it is used for irrigation and drinking purposes. Also, farmers use water from the rivers for agricultural purposes and the nearby areas peoples of the town, use both the surface and underground water as potable water, it is very harmful and unsafe to discharge this effluent into water body.

Here, present work is focused on the characterization of various disperse and vat dyes used by textile industries of Ludhiana for mutagenicity assessment.


Review of literature

Micro-organisms find to have much credit that makes them interesting for use in screening of effluents and chemicals for their toxicity. Approximately more than 200 tests preform on micro- organisms, insects, plants and animals, have been developed over the last 20-25 years, for the identification of chemical agents that cause genetic hazard to humans (Waters et al., 1988). The use or performance of bioassays is an essential part of the hazard estimation and their control procedures of the chemicals which are toxic to humans (Auletta et al., 1993 and Kirkland, 1993). Developed by B.N. Ames and co-workers in 1970’s (Ames et al., 1973 and 1975) the Salmonella assay has been used globally for nearly 4 decades, to estimate the mutagenicity of pure chemicals and complex environmental mixtures. The test was detects a large variety of mutagens, including those activated by mammalian liver enzymes.

The reverse mutation of Escherichia coli MTCC 40 tryptophan detects trp– to trpC reversion at a site blocking the biosynthesis of tryptophan previous to the formation of anthranilic acid. The MTCC 40 strains of E.coli all carry the same AT base pair at the mutation site within the trpE gene. Nowadays the assay is used by laboratories in conjunction with the Ames Salmonella assay for screening chemicals for checking mutagenic activity of these chemicals. In general terms the MTCC 40 strains are used as a substitute for, or as an addition to Salmonella strain TA102 which also have an AT base pair at the mutation site. The assay is also combine together with the Ames assay for data submission to regulatory agencies. National and international guidelines have been investigated for performing these mutagenicity assays. The E. coli WP2 assay procedures are the same as those described elsewhere in this volume for the Ames Salmonella assay (Mortelmans and Zeiger, 2000) with the exception that limited tryptophan instead of limited histidine is used.

The E. coli strain MTCC 40 notice that is a radiation resistant derivative of E. coli B/r which is the strain used by (Luria and Delbrück 1943) in their studies that estimate that mutations occur in bacteriato tryptophan and that mutations in the other strains arise during repair of DNA damage by the ‘error prone’ or ‘recombination’ repair system. Hill reported on the extreme sensitivity of strain e.coli MTCC40 uvrA to ultraviolet light irradiation (UV), as well as its susceptibility to enhanced reversion to tryptophan independence.


Excerpt out of 38 pages


Mutagenicity Assessment of Textile Dyes by Using Ames Test
Lovely Professional University, Punjab  (lovely professional university)
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ISBN (eBook)
ISBN (Book)
mutagenicity, assessment, textile, dyes, using, ames, test
Quote paper
Harpreet Kaur (Author), 2017, Mutagenicity Assessment of Textile Dyes by Using Ames Test, Munich, GRIN Verlag,


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