ISSN ONLINE(2319-8753)PRINT(2347-6710)

All submissions of the EM system will be redirected to Online Manuscript Submission System. Authors are requested to submit articles directly to Online Manuscript Submission System of respective journal.

Assessment of Antibacterial Activity of Bisphenol A (4,4’-Isopropylidenebisphenol).

Abdul Rasheed Md1, Ravi Kumar Kola2, Prameela Devi Yalavarthy3
  1. Research Scholar, Department of Zoology, Kakatiya University, Warangal, Andhra Pradesh, India
  2. Research Scholar, Department of Zoology, Kakatiya University, Warangal, Andhra Pradesh, India
  3. Professor, Department of Zoology, Kakatiya University, Warangal, Andhra Pradesh, India
Related article at Pubmed, Scholar Google

Visit for more related articles at International Journal of Innovative Research in Science, Engineering and Technology


Bisphenol A (BPA) is a chemical used to make polycarbonate plastics, epoxy resins, thermal paper. BPA-based products also include DVDs, computers, home appliances, spectacles and optical lenses, reusable water bottles, food storage containers, sports safety equipment, medical equipment, construction materials, paints and coatings. BPA contamination may occur from many different sources, but the most common way of BPA contamination is through packaged foods or beverages where the packing material contains BPA. The objective of this research paper is to evaluate the antimicrobial activity of Bisphenol A (4,4’-Isopropylidenebisphenol) against four different strains of bacteria i.e., Staphylococcus aureus, Bacillus substilis, Proteus valgaris and Escherichia coli, using agar diffusion method. BPA is completely soluble in organic solvents and partially soluble in water. The results showed the zone of inhibition in Staphylococcus aureus 23mm, Bacillus substilis 22mm, Proteus vulgaris 21mm and Escherichia coli 20mm. From the results it can be concluded that BPA may show the cytotoxic effects and these studies are also under way.


BPA, Bacillus substilis, Proteus vulgaris, Salmonella aureus, Escherichia coli, zone of inhibition and agar diffusion.


Bisphenol A (BPA) is the molecular building block for polycarbonate plastics and epoxy resins. U.S. production of BPA grew rapidly from 16 million pounds in 1991 to about 2.3 billion pounds in 2004, making it one of the most produced chemicals in the world [3]. BPA is a chemical recorded in the CAS (Chemical Abstract Service), number 00080-05-7 and its chemical name as used in Europe is 2, 2-Bis (4-hydroxyphenyl) propane [23]. There are a large number of synonyms to BPA like Bis(4-hydroxyphenyl)dimethyl methane; 4,4′-dihydroxydiphenyl propane; 4,4′-dihydroxy-2,2-diphenyl propane; Diphenylolpropane; 4,4′-isopropylidenediphenol. BPA is produced by the condensation of phenol and acetone in the presence of catalysts and catalyst promoters. Its molecular formula is C15H16O2 and molecular mass is 228.29 g / mol, and melting point is 155 °C (311 °F). The decomposition temperature is > 200°C (392°F) and it is completely soluble in organic solvents and partially soluble in water. It exists at room temperature in the form of a white solid flake or crystal [25]. BPA has been used in a wide variety of consumer products for several decades and continues to be manufactured in large quantities around the world. Humans are exposed to BPA through consumption of food and beverages contaminated with BPA, as well as environmental contamination. Polycarbonate plastic can become unstable over time and with use, allowing BPA to leach into material in contact with the plastics. Other consumer items, such as carbonless paper, computers, home appliances, spectacles and optical lenses, reusable water bottles, food storage containers, sports safety equipment, medical equipment, construction materials, paints, coatings and DVDs, also contain BPA. Additionally, BPA is now found nearly everywhere in the environment and commonly found in dust particles, surface water and drinking water, as over 6 billion pounds are produced worldwide each year [26]. Production of BPA releases approximately two hundred thousand pounds of the chemical into the atmosphere annually [19]. BPA in the rise of multiple diseases including prostate [5] and breast cancer [9], urino-genital abnormalities in male babies, a decline in semen quality in men [10], early onset of puberty in girls, metabolic disorders including insulin resistant (type 2) diabetes and obesity, neurobehavioral problems [1] such as attention deficit hyperactivity disorder (ADHD) and bisphenol-A also can change endogenous hormone synthesis, hormone metabolism and hormone concentrations in blood [27]. There are many reports on the contamination of BPA in the environment especially in water [28], air and soil [22, 17].


The technical grade quality of Bisphenol A is procured from HiMedia Laboratories Pvt. Ltd., (23 Vadhani Ind. Est., LBS Marg, Mumbai, India) for the study of antibacterial activity. The bacterial strains used in this study are Staphylococcus aureus, Bacillus substilis, which are gram positive bacteria and Proteus vulgaris and Escherichia coli which are gram negative bacteria. The bacterial strains were obtained from Microbial Type Culture Collection (MTCC), Institute of Microbial Technology (IMTECH), Chandigarh – India and maintained in the Microbiology Department of Kakatiya University. Two gram positive bacteria i.e.,Staphylococcus aureus and Bacillus substilis and two gram negative bacteria i.e.,Proteus vulgaris and Escherichia coli were used for assessing and evaluation of the antibacterial activity of the BPA. The method followed is agar well diffusion method [18, 24]. The test organisms were sub cultured using nutrient agar medium. The tubes containing sterilized medium were inoculated with respective bacterial strain. After incubation at 37±10C for 24 hrs they were stored in refrigerator. The stock culture was maintained. Bacterial inoculums were prepared by transferring a loop full of stock culture to nutrient broth. The flasks were incubated at 37±10C for 48 hrs before the experimentation.
Stock solution of the test compound was prepared by dissolving10mg/ml of DMSO and was used for testing at different concentrations. Streptomycin is used as standard drug for both Gram positive and Gram negative bacteria. The nutrient agar medium was sterilized by autoclaving at 1210C(12lbs/sq..inch) for 12 min. Petri-plates, tubes and flaks were sterilized in hot air oven at 1600C for an hour. In each sterilized petriplate (10cm diameter) about 27ml of molten nutrient agar medium which is inoculated with the respective strain of bacteria was poured. The plates were left at room temperature to allow solidification. In each plate seven discs of 6mm diameter were made with a sterile borer. The test compound, BPA at concentration 50μg/ml, 100μg/ml, 250μg/ml, 500μg/ml and 1000μg/ml was added to respective disc aseptically and labeled accordingly. The plates were kept undisturbed for 1 hour at room temperature to allow the diffusion of the solution properly in the nutrient agar medium. After incubation of the plates at 37±10C for 24 hours, the diameter of zone of inhibition surrounding each disc was measured with the help of an antibiotic zone reader. All the experiments carried out in triplicate. Simultaneously controls were maintained employing 0.1 ml of DMSO to observe the solvent effects and the results were represented in Table No.1.


As BPA is one of the highest volume chemicals produced worldwide, with over 6 million pounds produced each year [3] and is used for the production DVDs, computers, home appliances, spectacles and optical lenses, reusable water bottles, food storage containers, sports safety equipment, toys, water pipes, optical lenses, medical equipment, construction materials, paints and coatings, polycarbonate plastics, epoxy resins and thermal paper [5, 2]. Epoxy resins are used as protective linings for a variety of canned foods and beverages and as a coating on metal lids for glass jars and bottles. These uses result in consumer exposure to BPA via the diet [16]. Residues of BPA are also found in different environments such as rivers and wastewater effluents [14, 11]. Xenoestrogens are chemical with potent estrogenic properties [7]. BPA is classified as a xenobiotic disturbing hormonal balance in humans and other animals and it is an endocrine disruptor [20, 4] The exposure during early development to xenoestrogens such as BPA may be the underlying cause of the increased incidence of infertility, genital tract abnormalities, and breast cancer observed in European and US human populations [21]. Numerous toxicological and biochemical studies have confirmed that BPA has estrogenic properties and an agonistic effect towards the estrogenic receptor. The U.S. Food and Drug Administration (FDA) and the National Toxicology Program (NTP) have “some concern” for effects on the brain, behavior, and prostate gland in fetuses, infants, and children at current human exposures to BPA. Number of studies has examined BPA levels in other body fluids such as follicular fluid [12], urine [5] and semen [13, 15,]. BPA is found in milk due to contact with plastic materials during food processing and storage [6]. The Scientific Committee on Food (SCF), an advisory body of the European Commission on the safety of food, after comprehensive analysis of all aspects of BPA toxicity, has specified the tolerable daily intake (TDI) of BPA as 0.01 mg/ kg body mass per day [8]. Recent studies have shown that BPA can alter the gene expression (i.e. turned on or off) and the low-dose BPA exposure during pregnancy has multigenerational consequences and it may increase the likelihood of chromosomal abnormalities in F2 generation [26]. To test the effect of BPA as antibacterial agent, concentrations ranging from 0.05% to 1% were observed and the results are presented in [Table No.1] The antibacteriall activity of the BPA was studied by agar well diffusion method [18,24]. BPA is completely soluble in organic solvents and partially soluble in water [25]. DMSO scored as better solvent followed by methanol and ethanol, in terms of their compatibility with MIC determination. Hence it is dissolved in DMSO [29]. Among the different concentrations chosen i.e., 0.05%, 0.10%, 0.25%, 0.5% and 1%, BPA has shown strong antibacterial activity against all the bacterial strains that were tested [Fig.No.2]. It is found that the effect of BPA on the bacterial growth show the zone of inhibition in Staphylococcus aureus 23mm, Bacillus substilis 22mm, Proteus vulgaris 21mm and Escherichia coli 20mm [Table No.1]. BPA showed a maximum zone of inhibition in Staphylococcus aureus. and has antimicrobial activity against gram-negative and gram-positive bacteria. It has greater activity against gram-positive species over gram-negative species. It is evident from the study that the BPA is more effective in killing gram positive bacteria. The death of bacteria may be because it blocks lipid synthesis in them and has been shown to intercalate into bacterial cell membranes and disrupt membrane activities without causing leakage of intracellular components [30]. Since BPA is acting as xenoestrogen its usage should be limited in consumer products and especially the usage in child care products should be restricted. As there is a controversy and public concern in the use of BPA in consumer products as it possess hazardous health effects, methods should be developed for production of alternate compounds.


`From the above study it can concluded that the antibacterial activity increased with the increased concentrations of BPA It is evident from the study that the BPA is more effective in killing gram negative bacteria when compared to gram positive bacteria.


The authors wish to thank the Head, Department of Zoology, Kakatiya University, Warangal, for providing laboratory facilities. We thank to Department Microbiology, Kakatiya University., for providing bacterial strains to carry out this work.


1. Adriani W, Della Seta D, Dessi-Fulgheri F, Farabollini F and Laviola G”, Altered profiles of spontaneous novelty seeking, impulsive behavior and response to D-amphetamine in rats perinatally exposed to bisphenol A”. Environmental Health Perspectives Vol.111, pp.395-401, 2002.

2. Biedermann S, Tschudin P, and Grob K.”Transfer of Bisphenol A from thermal printer paper to the skin”, Analytical and Bioanalytical Chemistry. Vol.398, No.1, pp.571-6, 2010.

3. Burridge E. “Chemical profile Bisphenol A”. ICIS Chemical Business pp.274:48, 2008.

4. Boyd GR, Reemtsma H, Grimm DA., and Mitra S,. “Pharmaceuticals and personal care products (PPCPs) in surface and treated waters of Louisiana, U.S.A. and Ontario, Canada”, Sci Total Environ Vol.311, pp.135-149, 2003.

5. Calafat AM., Weuve J., Ye X., Jia LT., Hu H., and Ringer S., et al,. “Exposure to Bisphenol A and other phenols in neonatal intensive care unit premature infants”, Environmental Health Perspectives Vol.117(4), pp.639-644, 2009.

6. Casajuana N, Lacorte S. New methodology for the determination of phthalate esters, Bisphenol A, Bisphenol A diglycidyl ether, and nonylphenol in commercial whole milk samples. J Agric Food Chem. 2004 Jun 16;52(12):3702-7.

7. Dodds EC., Goldberg L., Lawson W., and Robinson R., “Estrogenic activity of certain synthetic compounds", Nature Vol.141(3562), pp.247-248, 1938.

8. EU, Commision of the European Communities 90/128/EU, Off. J. Eur. Community, L75, pp.19, 1990.

9. Hunt PA., Koehler KE., Susiarjo M., Hodges CA., IlaganA., Voigt RC., Thomas S., Thomas B., and Hassold T., “BisphenolAexposure causes meiotic aneuploidy in the female mouse”, Current Biology, Vol.13, pp.546-553, 2003

10. Hunt PA., Susiarjo M., Rubio C., and Hassold TJ., “The BisphenolA Experience: A primer for the Analysis of Environmental Effects on Mammalian Reproduction”, Biology of Reproduction, Vol.81, pp.807-813, 2009.

11. EFSA (European Food Safety Authority), “Bisphenol A for use in food contact materials” EFSA Journal, Vol.428, pp.10–75, 2009.

12. Ikezuki Y., Tsutsumi O., Takai Y., Kamei Y., and Taketani Y., “Determination of Bisphenol A concentrations in human biological fluids reveals significant early prenatal exposure”, Hum Reprod, Vol.17, pp.2839-41, 2002.

13. Inoue K., Wada M., and Higuchi T., et al., “Application of liquid chromatography-mass spectrometry to the quantification of bisphenol A in human semen”, Journal of Chromatography B, Vol.773, pp.97-102, 2002.

14. Brotons J.A., Olea-Serrano M.F., Villalobos M., PedrazaV., and Olea N., “Xenoestrogens released from lacquer coatings in food cans”, Environmental Health Perspectives, vol.103, No.6, pp.608–612, 1995.

15. Katayama M., Matsuda Y., Shimokawa K I., Ishikawa H., and Kaneko S., “Preliminary monitoring of Bisphenol A and nonylphenol in human semen by sensitive high performance liquid chromatography and capillary electrophoresis after proteinase K digestion”, Analytical Letters, Vol.36, pp.2659-67, 2003.

16. Kang, J.H., Kito, K., and Kondo, F. Factors influencing the migration of Bisphenol A from cans. Journal of Food Protection, Vol.66, No.8, pp.1444- 7, 2003.

17. Klecka GM., Staples CA., Clark KE., van der Hoeven N., Thomas DE., and Hentges SG., “Exposure Analysis of Bisphenol A in Surface Water Systems in North America and Europe”, Environ Sci Technol, Vol.43, No.16, pp.6145-6150, 2009.

18. Lyudmila Boyanova., Galina., Gergova., Rossena Nikolv., Sirigan derejian., Elena Lazarova, Nikolai Katsarov, Ivan Mitov and Zacharii Karastev., “Activity of Bulgarian propolis against 94Helicobacter pylori strains in vitro by agar well diffusion Methods:, J. Med. Microbial, Vol.54, pp.481- 483, 2005.

19. Markey CM., Michaelson CL., Sonnenschein C., and Soto AM., “Alkylphenols and Bisphenol A as environmental estrogens”, In Metzler M(Ed.),The Handbook of Environmental Chemistry. Part L, Endocrine Disruptors Part I, vol.3. Springer-Verlag, Berlin Heidelberg, pp.129– 153.2001

20. Moriyama K., Tagami T., Akamizu T., Usui T., Saijo M., and Kanamoto N., “Thyroid hormone action is disrupted by Bisphenol A as an antagonist”, J Clin Endocrinol Metab Vol.87, No.11, pp.5185-5190, 2002.

21. Munoz de Toro M M., Markey CM., and Wadia P R., “Perinatal exposure to Bisphenol A alters peripubertal mammary gland development in mice” Endocrinology, Vol.146, pp.4138-47, 2005.

22. Pingqing F., and K Kawamura., “Ubiquity of Bisphenol A in the atmosphere, Environmental Pollution”, Vol.158, pp.3138-3143, 2010.

23. Rykowska I., and Wasiak.W., “Properties, Threats, and Methods of analysis of Bisphenol A and its Derivatives”, Acta chromatographica, No.16, 2006.

24. Satish, S., Raveesha K.A., and Janartdhana G.R., “Antibactrial activity of plants extracts on Phytopathogenic Xanthomonas campestris pathovars” Letters in Applied Microbiology, Vol.28, pp.145-147, 2002.

25. Staples C A., Dorn P B., Klecka GM., O’Block S T., and Harris L R., “A review of the environmental fate, effects and exposures of bisfenol A” hemosphere PubMed, Vol.36, No.10, pp.2149-2173, 1998.

26. Susiarjo M., Hassold T J., Freeman E., and Hunt P A., “BisphenolAexposure in utero disrupts early oogenesis in the mouse”, PLoS Genetics, Vol.3, No.1, pp.63-70.2007.

27. Vom Saal F S., Cooke P S., Buchanan D L., Palanza P., Thayer K A., Nagel S C., Parmigiani S., Welshons W V., “A physiologically based approach to the study of bisphenol A and other estrogenic chemicals on the size of reproductive organs, daily sperm production, and behavior”, Toxicol Ind Health, Vol.14, No.1-2, pp.239-60, 1998.

28. Watanabe M., Hase T., and Imai Y., “Change in the bisphenol- A content in a polycorbonale orthodontic breacket and its leaching charastricts in Water”, Dent Mater J, Vol.20, pp358-58, 2001.

29. Wadhwani T., Desai K., Patel D., Lawani D., Bahaley P., Joshi P., and Kothari V., “Effect of various solvents on bacterial growth in context of determining MIC of various antimicrobials”, The Internet Journal of Microbiology, Vol7 Number 1. DOI: 10.5580/b43, 2009.

30. Knaysi G., and Morris G., “The manner of death of certain bacteria and yeast when subjected to mild chemical and physical agents”, Journal of Infectious Diseases 47, 303–17. (1930)