International Journal of Applied Pharmaceutics ISSN - 0975 - 7058 Vol 11, Special Issue 5, 2019 CYTOTOXIC ACTIVITY EVALUATION OF ERIOCAULON CINEREUM R.BR. ON HELA AND VERO CELL LINES PINUS JUMARYATNO 1 *, ARDE TOGA NUGRAHA 1 , ADILIA TRI HIDAYATI 1 , BAIQ RISKY WAHYU LISNASARI 1 , WIDYANUR MAYA DIAHANDARI 1 , NANANG FAKHRUDIN 2 1 Department of Pharmacy, Universitas Islam Indonesia, Yogyakarta, Indonesia. 2 Department of Pharmaceutical Biology, Faculty of Pharmacy, Universitas Gadjah Mada, Yogyakarta, Indonesia. Email: pinus.jumaryatno@uii.ac.id Received: 19 December 2018, Revised and Accepted: 23 January 2019 ABSTRACT Objective: This study aimed to evaluate the cytotoxic activity of the extracts and fractions of Eriocaulon cinereum against HeLa and Vero cell lines, which represent cervical cancer and normal cells, respectively. In addition, a phytochemical screening was carried out to determine the chemical constituents in the extracts and the active fractions. Materials and Methods: The extracts of E. cinereum were obtained by ultrasound-assisted extraction method using n-hexane, ethyl acetate, and methanol, successively. The active extract was fractionated using vacuum liquid chromatography with dichloromethane followed by ethyl acetate. The cytotoxic activity was evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay method and was measured using microplate reader at the wavelength 595 nm. The data were analyzed with PROBIT from SPSS 16 for Windows ® . In addition, phytochemical screening was performed using standard procedures. Results: The cytotoxic evaluation of the extracts of E. cinereum showed that the ethyl acetate extract was the most active extract against HeLa cell line with the half maximal inhibitory concentration (IC 50 ) value of 580.07 μg/ml. The dichloromethane and ethyl acetate fractions from the active extract of E. cinereum exhibited cytotoxic activity against HeLa cell with the IC 50 values of 466.61 μg/ml and 267.34 μg/ml, respectively. In addition, the ethyl acetate fraction showed a low cytotoxic effect against Vero cell line. The phytochemical screening of the ethyl acetate fraction indicated the presence of terpenoids and alkaloids. Conclusion: This finding revealed the anticancer potential of E. cinereum and warranted further investigation for the discovery of new anticancer agents from natural resources for cervical cancer. Keywords: Cytotoxic, Eriocaulon cinereum, HeLa cell line, Phytochemical screening, Vero cell line. INTRODUCTION Cervical cancer is a cancer commonly caused by the human papilloma virus. The World Cancer Report 2014 reported that cervical cancer is the fourth most common cause of death in women affected by cancer [1]. While in Indonesia, according to the data from the Ministry of Health of the Republic of Indonesia, the prevalence of cervical cancer is 0.8% of the total population of Indonesia [2]. Therefore, cervical cancer is one of the priorities for the Indonesian Ministry of Health in terms of combating cancer. Until now, the most common therapy for patients with cervical cancer is a surgery together with a radiotherapy for the early stage of cancer or a combination of radiotherapy and chemotherapy for advanced stage. The drugs commonly used for chemotherapy in the treatment of cervical cancer include cisplatin, carboplatin, paclitaxel, topotecan, and gemcitabine [3-7]. However, chemotherapy will cause unwanted side effects such as nausea, vomiting, hair loss, mouth sores, fatigue, and appetite loss [8]. Nature is known to have a rich source of natural compounds that have the health benefits to treat cancer, including the plant from Eriocaulon genus. Previous reports showed that some Eriocaulon species have the cytotoxic activity. For example, Eriocaulon australe from China has cytotoxic activity against A549 lung cancer cells, MCF-7 breast cancer cells, and HeLa cervical cancer cells [9]. Meanwhile, Eriocaulon sieboldianum was reported to exhibit cancer activity against HL-60 human leukemia cell line [10] and induce cell cycle arrest of human liver cancer cells HepG2 [11]. Furthermore, Fan et al. also reported antitumor activity against K562 leukemia cells from E. sieboldianum [12]. In addition, E. sieboldianum has been used as adjuvant therapy to treat cervical cancer in traditional Chinese medicine [12]. Nugraha et al. [13] conducted a preliminary study to screening the Eriocaulon cinereum R.Br. from Bangka Belitung Island, Indonesia, for anticancer. This plant has been used by local people to prevent and treat uncontrolled cells growth. The result showed the inhibition of HeLa cells growth with the half maximal inhibitory concentration (IC 50 ) value of 428 µg/mL on ethanol extract [13]. Based on this result, further investigation of E. cinereum was carried out as a source for new anticancer therapeutics. Therefore, the aims of this study were to investigate the cytotoxic activity of the extracts of E. cinereum and its fractions against HeLa cells and to identify the phytochemicals present in the active extract and fractions. MATERIALS AND METHODS Plant materials The plant was collected from Parittiga, Jebus, Bangka Belitung Island Province, Indonesia, on July 2016. E. cinereum R.Br. was identified by Drs. Heri Sujadmiko and MSi. from the Laboratory of Plant Systematic at Faculty of Biology, Universitas Gadjah Mada, Yogyakarta, Indonesia. Voucher specimen was deposited at Laboratory of Pharmaceutical Biology, Department of Pharmacy, Universitas Islam Indonesia, Yogyakarta, Indonesia (Collection No. BF-FAUII-01). Samples were dried using cabinet dryer at 50°C for 36 h and powdered. © 2019 The Authors. Published by Innovare Academic Sciences Pvt Ltd. This is an open access article under the CC BY license (http://creativecommons. org/licenses/by/4. 0/) DOI: http://dx.doi.org/10.22159/ijap.2019.v11s5.T0059 4 th International Conference on Pharmacy and Pharmaceutical Science (ICPPS) 2019 Research Article