Thematic Special Issue: Modern Drug Discovery – Current Challenges & Future Perspectives 2022 | 84 Original Article GCMS ANALYSIS ON ANDROGRAPHIS PANICULATA SEED EXTRACT AND ITS ANTICANCER ACTIVITY RAJESWARI S., VIDYA, R. * , AMUDHA P. * Department of Biochemistry, School of Life Sciences, Vels Institute of Science, Technology and Advanced Studies (VISTAS), Chennai, Tamil Nadu, India * Email: vidyabiochem@gmail.com Received: 16 Apr 2021, Revised and Accepted: 30 May 2022 ABSTRACT Objective: Andrographis paniculata was an ancient medicinal herb recorded in Ayurvedic system of medicine from long times. The present study is to determine the bioactive phytocomponents present in Andrographis paniculata seed, extraction done using ethanol and Human Liver Cancer (HepG2) cells are used to prove the anticancer activity. Methods: Gas Chromatography-Mass Spectroscopy (GCMS) analysis is used for determining the bioactive constituents of the Andrographis paniculata seed. To prove the in vitro anticancer activity, the MTT assay was done on Human Liver Cancer (HepG2) cells. Results: GCMS analysis of Andrographis paniculata seed extract result determined the presence of 15 active bio components. These various potent bioactive compounds have been proved to possess various pharmacological activities, viz., antimicrobial, antidiabetic, antioxidant, anti- inflammatory and anticancer properties, which were supported by previous findings. The natural compounds with therapeutic effects on traditional medicine have paved the way for evaluating the Andrographis paniculata seeds for cytotoxic activity on HepG2 cell lines. The ethanol extract of Andrographis paniculata seeds was used on HepG2 to evaluate the cytotoxicity; Cisplatin (15 µg/ml) is used as standard with various concentration like 200, 100, 50 and 25 µg/ml were used. The bioactive compounds present in Andrographis paniculata seed extract revealed significant cytotoxicity activity against HepG2 cells with IC50 value of 103.03µg/ml. Conclusion: The cytotoxic effect was found to be concentration-dependent and increased concentration of Andrographis paniculata seed extract showed increased cytotoxicity. The result suggest Andrographis paniculata seed extract have bioactive agents which when treated on hepatocellular carcinoma cells poses excellent anticancer activity. Keywords: Bioactive components, GCMS, Anticancer, Hepatocellular carcinoma cells, Cytotoxicity © 2022 The Authors. Published by Innovare Academic Sciences Pvt Ltd. This is an open access article under the CC BY license (https://creativecommons.org/licenses/by/4.0/) DOI: https://dx.doi.org/10.22159/ijap.2022.v14ti.5 Journal homepage: https://innovareacademics.in/journals/index.php/ijap INTRODUCTION Medicinal plants have profound therapeutic applications and has become an integral part of human life, which could combat the sufferings due to various diseases. The pharmaceutical industries are now focusing on indigenous production of drugs using these medicinal plants. In Ayurvedic formulation, Andrographis paniculata or Kalmegh is one of the most widely used plants. Occurrence of Andrographis paniculata is common in most of the place in India, unlike other species. In hilly areas it grows up to 500m and its of wide use; communities in India have been using this herb for treating variety of diseases [1]. The phytocomponents likes diterpenoids, lactones, flavonoids, and flavonoid glycosides are found in aerial part of Andrographis paniculata’s extract [2]. Andrographolide is quantitatively the major bioactive secondary metabolite present in this herb. The antioxidant, anti-carcinogenic, antiparasitic, antiparasitic, antiHIV, antiHIV, anti- bacterial, anti-inflammatory, anticancer etc are the activities possessed by Andrographis paniculata which are reported in previous studies [3]. The death caused from Malaria, HIV/AIDS, Tuberculosis is less than the death caused by cancer and it rates about 8.8 million people every year. In 2035, the incidence of cancer would increase double times [4]. Among different cancer, rates of liver cancer in Africa and Asia are found to be high. The chronic infections with Hepatitis C virus (HCV) and Hepatitis B virus (HBV) are the causes of Hepatic Cellular Carcinoma (HCC) with a percentage of about 73.4% [5]. Andrographis paniculata leaf extract poses anticancer activity against H-29 cancer cell, which shows 50% inhibition at 200 μg/ml concentration for ethanol extract, proves to be an alternative medicine for cancer [6]. In Ovcar-5 cell line the inhibition was 51.12% when exposed to hydroalcoholic extract of Andrographis paniculata. In HepG2 cell the inhibition rate was 42.76% when exposed with a combination of Andrographis paniculata and S. marianum extract [7]. Ethanol extract of Andrographis paniculata can inhibit MCF-7 cell and studies are need on neoandrogpholide inhibition of adenylate kinase 2 in MCF-7 cells [8]. Due to the beneficial medical properties of Andrographis paniculata the demand is getting increased day by day. Many research report is available on the biological and pharmacological aspects but none of the reports deals with Andrographis paniculata seeds [9]. The present investigation is considered as important as it is the first of its kind. No previous reports are available on the medicinal use of Andrographis paniculata seeds. MATERIALS AND METHODS Extraction of bioactive components The Andrographis paniculata seeds were purchased from local market, Avadi, Tamil Nadu. The seed samples were wild varieties, which are not macro or micro, propagated by human. The seeds were identified registration number of the certificate: PARC/2020/4258 which was authenticated by Dr. P. Jayaraman, Director, Plant Anatomy research Centre, Tambaram. The seeds of Andrographis paniculata were dried in shade, cleaned and powdered with blender. In Soxhlet apparatus, 20 ml of ethanol (V/V) were taken and 50 g of powder sample of Andrographis paniculata were added. By using rota evaporator, the extract was filtered, and the content is evaporated for drying and finally, it is stored in the refrigerator at 2-8 °C. The final seed extract obtained are used for identification of phytoconstituents which are present in the extract [10]. Gas chromatography-mass spectroscopy (GCMS analysis) The analysis in ethanol extract of Andrographis paniculata seeds for phytocompounds are performed using GCMS technology. The Perkin Turbo gold mass detector was equipped with a fused silica column containing BR-5MS (5% Diphenyl) with Andrographis paniculata seeds extract of 2 µl was used for GCMS analysis. The flow rate was International Journal of Applied Pharmaceutics ISSN- 0975-7058 Thematic Special Issue 2022