Research Articles

2019  |  Vol: 5(4)  |  Issue: 4 (July- August)  |
Cytotoxicity activity of Erythrina indica flowers and separation of essential oil

S. Vijaya Bharathi*, Gargi Srivastava

Department of Biotechnology

FSH, SRMIST, Kattankulathur, Tamilnadu- 603203, India

*Address for Corresponding Author

S. Vijaya Bharathi

Department of Biotechnology

FSH, SRMIST, Kattankulathur, Tamilnadu- 603203, India


Objective: To study the anticancer potential and separation of essential oil from flowers of Erythina indicaMaterial and methods: Antibacterial efficiency was checked by well diffusion method. In to the prepared Muller Hinton agar plates, test cultures were swabbed, incubated for 24 hours. Then well was made and added different concentrations of extract. Cytotoxicity test was done by adding various concentration of extract to the cell lines, incubated and finally MTT solution was added. Formazan formation was measured at 550nm. Cell morphology was checked by microscopy analysis and steam distillation was carried out to separate essential oil. Results and conclusion: Among the 5 test organism analysed, extract showed maximum zone of inhibition for S. aureus. It was identified that the flower ethanol extract was effective against both the cell lines with the cytotoxicity % of 71.26 towards HepG2 and against MG-63 showed 67.11. Hydrosol and the essential oil have to check for its chemical composition, pharmaceutical applications.

Keywords: Erythrina indica, antibacterial, HepG2, hydrosol


Globally utilization of plants as therapeutics agent is increasing. At the same time when a plant is located as valuable drug, its population become prone to wild crafting and unsustainability. Considering this WHO (World health organisation, 2003) and European Medicines Agency (EMA, 2006) developed guidelines for collection and sustainability of medicinal plants (Atanasov et al., 2015). Vegetarian foods are considered as super nutrient source, as it contains flavanoids, alkaloids and other pharmaceutically important compounds (Shakya, 2016). In this study, cytotoxicity potential of and essential oil extraction of Erythrina indica (EI) flower was carried out to explore its medicinal importance.

Erythrina indica is a fabaceae plant commonly found in tropical and subtropical regions of world. This plant can grow up to a height of 30- 60 meters. It can be referred as coral tree commonly as it holds red colour flower. These are coral trees peculiarly cultivated for their ornamental flowers, they play major role in nitrogen fixation in soil. The plant is commonly known as ‘Kalyana murungai’ in Tamil language (Lahari et al., 2015). Flowering is continued by seed production and they are brown in colour (Nagar and Chauhan, 2015).

IARC, 2018 (12th September) release states that new cancer affected cases reached 18.1 million and 9.6 million deaths. Also it was identified that the lung cancer, female breast cancer and colorectal cancer are more prevalent. In men, the leading cause for the death is lung cancer which followed by prostate cancer, colorectal, liver and stomach cancer. Likewise in women the breast cancer is the most commonly diagnosed, followed by lung, colorectal and cervical cancer (

Liver cancer is recorded as third leading cause of cancer associated death. Liver trans-plantation, surgical resection and chemotherapy are the common treatment practice of liver cancer. But the chemotherapeutic agents available are not effective in advanced stage of cancer and so the plant based drugs which can work in such case will be boon to the society ( Jiao et al., 2018). Mostly children and young adults are affected by osteosarcoma. Like in other cases of cancer, bone cancer also treated surgical or by chemotherapy (Jirangkul et al., 2014). As there is a large need of plant based agent to treat cancer, the present study is focussed on assessing the anticancer potential of EI flower.

Materials and methods

Ethanol extraction of Erythina indica (EEI)

Fresh flowers of Erythrina indica was collected from local place of Chennai city washed and dried in shade for 1 week and authenticated. Shade drying is the traditional method commonly followed for drying the plant material (Dwivedy et al., 2012). 10g of EI flower was transferred to the conical flask containing 100ml of ethanol, placed in orbital shaker with for 7 days, filtered, evaporated and stored (Okoduwa et al., 2016).

Metabolite analysis

Preliminary analysis of secondary metabolites was carried out by Trease and Evans procedure (Trease and Evans, 2009).

Antibacterial activity

Culture plates were made by pouring Muller Hinton agar to the depth of 3-4mm, allowed to dry. Then the test culture (E. coli, A. hydrophilia, K. pneumonia, B. cereus and S. aureus) was swabbed evenly on to the agar and incubated for 24hrs at 37°C. Wells were made of 5mm diameter. 30 µl of three different concentrations (10µg/ml, 20µg/ml and 30µg/ml) of EEI and ampicillin were added to pre-marked wells. After 24 hrs incubation the zone of inhibition was measured (Yadav et al., 2015).

Cell culturing and cytotoxicity assay

Cell lines were properly cultured in DMEM (Dulbecco’s modified eagle medium) and grown overnight (96- well plate) at 37˚C in a 5% CO2 incubator. EEI of five different concentration (20, 40, 60, 80 and 100 µg/ml) were added, incubated for 24hrs. Then 50 μL of 2 mg/mL MTT solution was added to the well and incubated for 4hrs. After incubation, all supernatant was discarded and 100 μL of dimethyl sulfoxide was added to dissolve the formazan crystals. Absorbance was measured at 550nm and cell viability was calculated. MTT is cleaved by mitochondria dehydrogenase in viable cells, giving a measurable purple product formazan. This formazan production is directly proportionate to the viable cell number and inversely proportional to the degree of cytotoxicity (Han et al., 2015). Morphological change was observed under inverted microscope (Husni et al., 2015).

Steam distillation

Essential oil was extracted by steam distillation procedure. Essential oil has applications in pharma, agricultural, food and perfume industries (Gakuubi, 2016).

Results and discussion

Alkaloids and phenolic compounds like tannins, flavanoids were showed positive in EEI (Table 1). Different solvents extract of EI leaf revealed the same kind of secondary metabolites as in EEI (Kumari et al., 2017).

Table 1. Secondary metabolites of EEI

S. No

 Test for bioactive compounds

Erythrina indica ethanol extract


Alkaloids test



Flavonoids test



Saponins test



Phenolic compound test



Steroids test



Tannins test





+ positive, - negative.

EEI was found to be highly effective against S. aureus with 6.1mm as maximum zone of inhibition, whereas against K. pneumoniae it showed 5.4mm, E.coli of 5.1mm and B.cereus with 2.6mm. A. hydrophilia showed the lowest zone of inhibition (Figure 1, 2 and 3). Stem methanolic extract of EI was found to be effective against E. coli (18mm) comparable to S. aureus (10mm) (Sahoo et al., 2012). Similarly aqueous leaf extracts was also have high inhibitory activity against E. coli than towards S. aureus as in case of methanol extract of EI stem (Kumari et al., 2017). Ethanol leaf analysed by Christy Jeyaseelan et al. (2017) revealed that the extract has potential inhibition against E. coli but S. aureus showed less inhibition potential (Jeyaseelan et al., 2017).

Figure 1. Zone of inhibition of E. coli, A. hydrophilia


Figure 2. Zone of inhibition of (a) K. pneumoniae (b) B. cereus (c) S. aureus




Figure 3. Antibacterial activity of Erythina indica


Cytotoxicity potential was dose dependent against HepG2 and MG63. EEI showed 71.26% of cytotoxicity against HepG2 whereas 67.11% cytotoxicity towards MG63 (Table 2). Thorough literature review reveals that the leaves and the other parts of EI has checked for anticancer potential. EI leaf ethanol extract against adenocarcinoma results showed 78.89% cytotoxicity at 1000µg/ml (Priya et al., 2017). 10, 11-dioxoerythratidine and the crystagallin A were the two main compounds of EI leaf and stem showed potential anticancer activity against breast cancer T47D cell- line (Herlina et al., 2011). Morphology analysis revealed that EEI presence caused cell shrinkage which was not seen in control (Figure 4a, b, c, d). Breast cancer cell line assessed with aqueous Erythrina indica extract also showed similar morphological change when studied with acridine orange and Giemsa stains (Rai et al., 2017). Further analysis of pure compound responsible for anticancer activity is needed.  

Table 2. Cell toxicity assay

Concentrations (µg/mL)

Cell toxicity (%) HepG2

Cell toxicity (%) MG-63



















Figure 4.  Cytotoxicity  effect of (a) Control (HepG2); (b) EEI (HepG2); (c) Control (MG-63); (d) Cytotoxicity of EEI (MG-63)



Essential oil extracted was separated from hydrosol by freezing method. 1ml of essential oil was collected. Both hydrosol and essential oil have wide application and so the constituents of the obtained oil, hydrosol has to be studied in detail in future. Essential oils are widely utilized because of it limited side effects (Mahmoudi, 2017). Hydrosols of Lavender showed efficient antimicrobial activity as well as a potential preservative in moisturizing body gel (Styczynska et al., 2014).


Erythrina indica flower is commonly known for its ornamental purpose but its medicinal value observed showed that it can be utilized for drug development by carrying out in-vivo study and also pure bioactive compound isolation can be done for effective results.


We would like thank Department of Biotechnology, FSH, SRMIST, for providing as the lab facility.

Conflict of interest

There is no conflict of interest.


Atanasov AG , Waltenberger B , Pferschy-Wenzig EM , Linderd T, Wawroscha C, Uhrine P, Temml V, Wanga L, Schwaigerb S, Heissa EH, Rollingera JM, Schusterf D, Breusse JM, Bochkovg V, Mihovilovicd MD, Koppa B, Bauerc R , Dirscha VM, Stuppnerb H. 2015. Discovery and resupply of pharmacologically active plantderived natural products: A review. Biotechnology Advances 33(8):1582–1614. 

Dwivedy S, Rayaguru K, Sahoo GR. 2012. Effect of Drying Methods on Quality Characteristics of Medicinal Indian Borage (Coleus aromaticus) Leaves. Journal of Food Processing and Technology 3:188.

Gakuubi MM. 2016. Steam distillation extraction and chemical composition of essential oils of Toddalia asiatica L. and Eucalyptus camaldulensis Dehnh.  Journal of Pharmacognosy and Phytochemistry 5(2):99-104.

Han BJ, Li W, Jiang GB, Lai SH, Zhang C, Zeng CC, Li YJ. 2015. Effects of daidzein in regards to cytotoxicity in vitro, apoptosis, reactive oxygen species level, cell cycle arrest and the expression of caspase and Bcl-2 family proteins.  Oncology Reports 34:1115-1120.

Herlina T, Supratman U, Subarnas A, Sutardjo S, Amien S,  Hayashi H. 2011. In Vitro Anti-Cancer Alkaloid and Flavonoid Extracted from the Erythrina variegata (Leguminoseae) plant.  Indonesian Journal of Cancer Chemoprevention 2(3):286-290.

Husni E, Nahari F, Wirasti Y, Wahyuni FS, Dachriyanus. 2015. Cytotoxicity study of ethanol extract of the stem bark of asam kandis (Garcinia cowa Roxb.) on T47D breast cancer cell line.  Asian Pacific Journal of Tropical Biomedicine 5(3):249-252.

Jeyaseelan EC, Jesuthasan J, Tharmila CJ, Jashothan PTJ, Jeyadevan JP. 2017. Authentication of Antibacterial Activity of Wound Healing Siddha Medicinal Plants. Der Pharma Chemica 9(15):5-8.

Jiao Y, Wu Y, Du D. 2018. Polydatin inhibits cell proliferation, invasion and migration, and induces cell apoptosis in hepatocellular carcinoma.  Brazilian Journal of Medical and Biological Research 51(4):1-9.

Jirangkul P, Srisawat P, Punyaratabandhu T, Songpattanaslip T, Mungthin M. 2014. Cytotoxic effect of artemisinin and its derivatives on human osteosarcoma cell lines.  Journal of the Medical Association of Thailand 97 (Suppl.) 2. 

Kumari P, Singh SK, Kumari C. 2017. Phytochemical Screening and Antibacterial Activity of Erythrina variegata L. (Leaf Extract). International Journal of Current Microbiology and Applied Sciences 6(6):2500-2505.

Lahari K, Divya M, Vidyavathi N, Kishore L, Poojitha M. 2015. Novel review on Erythrina variegata. International Research Journal of Pharmacy 6(4):231-233.

Mahmoudi R. 2017. Application of medicinal plants: from past to present. MedCrave Online Journal of Biology and Medicine 1(3):80.

Nagar JC and Chauhan LS. 2015. Hypoglycemic and hypolipidemic activity of root extracts of Erythrina variegate in alloxan induced diabetic rats.  Asian Journal of Biomedical and Pharmaceutical Sciences 5(46):25-30.

Okoduwa SIR, Umar IA, James DB, Inuwa HM, Habila JD. 2016. Evaluation of extraction protocols for anti-diabetic phytochemical substances from medicinal plants. World journal of Diabetes 7(20):605-614.

Priya R, Mani P, Maneemegalai S. 2017.  Anti-Oxidant and Anticancer Properties of Ethanol Leaves Extract Of Erythrina Indica. European Journal of Biomedical and Pharmaceutical sciences 5(1):438-443.

Rai MV, Pai VR, Kevin S, Kedilaya HP. 2017. In Vitro Evaluation of Anticancer Potential of Erythrina Variegata L. On Breast Cancer Cell Lines. Asian Journal of Pharmaceutical and Clinical Research 10(7):305-310.

Sahoo K, Panda SS, Das D, Dhal NK. 2012. In Vitro Analysis Of Antimicrobial Activity Of Stem Extracts Of Erythrina Variegata L: A Useful Medicinal Plant. International Journal of Pharma and Bio Sciences 3(3):766–772.

Shakya AK. 2016. Medicinal plants: Future source of new drugs. International Journal of Herbal Medicine 4(4):59-64.

Styczynska AK, Smigielski K, Prusinowska R , Rajkowska K , Kusmider B, Sikora M. 2014. Preservative activity of lavender hydrosols in moisturizing body gels. Letters in Applied Microbiology 60:27-32.

Trease and Evans, 2009. Pharmacognosy. 16th Ed, Saunders Ltd.

Yadav S,  Trivedi NA,  Bhatt JD. 2015. Antimicrobial activity of fresh garlic juice: An in vitro study.  Ayu  36(2):203–207.

Manuscript Management System
Submit Article Subscribe Most Popular Articles Join as Reviewer Email Alerts Open Access
Our Another Journal
Another Journal
Call for Paper in Special Issue on

Call for Paper in Special Issue on