Journal of Applied Pharmaceutical Science Vol. 8(08), pp 012-020, August, 2018 Available online at http://www.japsonline.com DOI: 10.7324/JAPS.2018.8802 ISSN 2231-3354 © 2018 Haryati Ahmad Hairi et al. This is an open access article distributed under the terms of the Creative Commons Attribution License -NonCommercial- ShareAlikeUnported License (http://creativecommons.org/licenses/by-nc-sa/3.0/). * Corresponding Author Ahmad Nazrun Shuid, Department of Pharmacology, Faculty of Medicine, Pre clinical Building, Universiti Kebangsaan, Malaysia, Jalan Yaacob Latiff, Bandar Tun Razak, Cheras, 56000 Kuala Lumpur. E-mail: anazrun @ yahoo.com Demethylbelamcandaquinone B Isolated from Labisia Pumila Haryati Ahmad Hairi 1 , Jamia Azdina Jamal 2 , Nor Ashila Aladdin 2 , Khairana Husin 2 , Noor Suhaili Mohd Sof 1 , Norazlina Mohamed 1 , Isa Naina Mohamed 1 , Ahmad Nazrun Shuid 1* 1 Department of Pharmacology, Faculty of Medicine, Preclinical Building, Universiti Kebangsaan Malaysia, Jalan Yaacob Latiff, Bandar Tun Razak, Cheras, 56000 Kuala Lumpur, Malaysia. 2 Faculty of Pharmacy, Universiti Kebangsaan Malaysia, Jalan Raja Abdul Aziz, 50300 Kuala Lumpur, Malaysia. ARTICLE INFO ABSTRACT Article history: Received on: 19/05/2018 Accepted on: 19/07/2018 Available online: 31/08/2018 (LP) or more commonly known as Kacip Fatimah in Malaysia has received much attention due to its estrogenic effects, including its role in the treatment of osteoporosis. This study was designed to explore the active compound of LP that may be responsible for its anti-osteoporotic effects. Crude aqueous extract of var alata (LPva) was fractionated into hexane (Hex), dichloromethane (DCM) and methanol (Met) solvents and their proliferative effects on mouse osteoblastic cell line (MC3T3-E1) were evaluated with MTS bioassay. The DCM fraction signifcantly promoted cell proliferation in a dose-dependent manner. Thin layer chromatography (TLC) was performed on the DCM fraction of LPva to separate the constituents and the potential active compound was identifed. Further isolation was achieved by column chromatography (CC) and Sephadex LH-20 column chromatography. The bioactivity of the isolated compound was confrmed by its ability to replicate MC3T3-E1 accelerated proliferation and differentiation. Mass spectrometry and nuclear magnetic resonance (NMR) identifed the active compound as demethylbelamcandaquinone B. Further studies are required to determine the potential of this active compound of LPva in treating osteoporosis. Key words: active compound, Labisia pumila, osteoporosis, osteoblasts. INTRODUCTION Osteoporosis, a worldwide prevalence of disease, can be defned as a systemic skeletal disease characterized by weak bones and high risk of fractures (Kanis and Johnell, 2006). This disease generally affects the elderly population and is associated with postmenopausal estrogen defciency and progressive bone loss (McCann et al., 2008; Pisani et al., 2016). Low estrogen level leads to excessive bone resorption and reduced bone formation (Bord and Beavan, 2003; Sorensen et al., 2006). The imbalance of bone remodeling resulted in a bone loss and eventually osteoporosis (Feng and Mcdonald, 2011). The main option for prevention and treatment of postmenopausal osteoporosis is hormone replacement therapy (HRT). However, HRT was linked to breast carcinoma (Kotsopoulos et al., 2016), endometrial cancer (Chlebowski et al., 2016) and cardiovascular disease (Bassuk and Manson, 2016). Bisphosphonate is a synthetic drug used to reduce the risk of vertebral and hip fractures, but daily bisphosphonate therapy may cause esophagitis and osteonecrosis of the jaw (Xu et al., 2013). Selective estrogen receptor modulator or SERM is another synthetic drug that binds to the intracellular estrogen receptors in target organs as agonists or antagonists. Unfortunately, it may also cause serious side effects such as thromboembolic disorders and hot fushes (Maximov et al., 2013). Recently, phytoestrogen became a favorable alternative therapeutic agent for treatment of osteoporosis. Phytoestrogens, such as isofavones (Deyhima et al., 2003; Singh et al., 2015), lignans (Zhu et al., 2017) and coumestans (Ueda et al., 2011) acted as SERMs to exert therapeutic effects against postmenopausal osteoporosis. These phytoestrogens could also promote calcium absorption or regulate bone remodeling through estrogen receptor Labisia pumila Labisia pumila Enhanced Proliferation and Differentiation of Osteoblast Cells Labisia pumila