Journal of Applied Pharmaceutical Science Vol. 10(12), pp 075-082, December, 2020 Available online at http://www.japsonline.com DOI: 10.7324/JAPS.2020.101210 ISSN 2231-3354 Cytotoxicity and chromatographic analysis of Dioon spinulosum, family Zamiaceae Marwa Elghondakly 1 , Abeer Moawad 2* , Mona Hetta 3 1 Pharmacognosy Department, Faculty of Pharmacy, Nahda University, Beni-Suef, Egypt. 2 Pharmacognosy Department, Faculty of Pharmacy, Beni-Suef University, Beni-Suef 62514, Egypt. 3 Pharmacognosy Department, Faculty of Pharmacy, Fayoum University; Fayoum, 63514, Egypt. ARTICLE INFO Received on: 23/04/2020 Accepted on: 13/09/2020 Available online: 05/12/2020 Key words: Dioon spinulosum, endocervix carcinoma, breast cancer, biflavonoids, aromadendrin, DNA analysis. ABSTRACT The identification of cytotoxic secondary metabolites from Dioon spinulosum Dyer ex. leaves was our aim. Thus, the evaluation of the cytotoxic activity of the total alcohol extract and successive fractions [n-hexane, dichloromethane (DCM), ethyl acetate, and n-butanol] against endocervix carcinoma (HeLa) and breast cancer (MCF7) cell lines was carried out using the sulforhodamine B assay. Identifying and authenticating D. spinulosum Dyer by DNA fingerprinting were carried out using Start Codon Translation (SCoT) and Inter simple Sequence Repeat, and revealed that SCoT4, SCoT6, SCoT8, HB-9, and HB-14 can be used for the identification of D. spinulosum at the genetic level. Chromatographic analysis and isolation, followed by the spectroscopic detection of the isolated compounds, were achieved by using various spectroscopic techniques. The following five compounds were isolated: β-sitosterol (1), 7,7'',4',4'''-O-tetra-methylamentoflavone (2), sciadopitysin (3), amentoflavone (4), and aromadendrin (5). Compounds 1 and 5 were obtained for the first time from the titled plant. This plant showed good cytotoxic activity against endocervix carcinoma as the total ethanolic extract, n-hexane, DCM, ethyl acetate, and n-butanol fractions showed IC 50 values of 21.8, 21.1, 24.5, 26.5, and 21.4 µg/ml respectively, against HeLa cell lines. On the other hand, the total ethanolic extract and the fraction of ethyl acetate showed good anticancer activity against MCF7 cell line with IC 50 values of 22.5 and 18 µg/ml, respectively. Aromadendrin showed IC 50 = 12.5 µg/ml against MCF7 cell line. In conclusion, aromadendrin may be responsible for D. spinulosum cytotoxicity against breast cancer cell lines. The total extract exhibited a promising adjunctive therapy for anticancer patients with endocervix and breast carcinomas. INTRODUCTION Plant-derived compounds have an undeniable role in the treatment of various diseases, including cancer. Several approved antitumor drugs have been extracted from plants, and several other compounds are in cancer clinical trials (Tayarani- Najaran and Emami, 2011). Many studies have showed that plant species are being used for cancer treatment or prevention of cancer development, especially in developing countries (Fouché et al., 2008; Ochwang’I et al., 2014). Cycads are plants belonging to class Gymnospermae, order Cycadales. The seeds of cycads in many parts of the world are used in the form of flour to produce many edible products (Nair and Van-Staden, 2012). Dioon genus is one of the cycads that belongs to the family Zamiaceae and is represented in Egypt by only two species: D. spinulosum Dyer ex. Eichler and D. edule Lindl (Hamdy et al., 2007). D. spinulosum Dyer ex. Eichleris is native to Mexico and South Africa and superficially resembles a palm (Johnson and Wilson, 1990). Being a member of cycads, Dioon genus has biflavonoids as secondary metabolites (Harborne, 1977). Amentoflavone-type biflavonoids, such as amentoflavone, bilobetin, sequoiaflavone, isoginkgetin, sciadopitysin, 7,4′,7″,4‴-tetra-O-methyl amentoflavone, and diooflavone (amentoflavone hexamethyl ether), were previously reported (Dossaji et al., 1975). It also has C-glycosylflavones, which were detected by Thin Layer Chromatography (TLC) (Carson and Wallace, 1972). On the other hand, hinokiflavone and its derivatives is absent from the whole family (Meurer and Stevenson, 1994). * Corresponding Author Abeer Moawad, Pharmacognosy Department, Faculty of Pharmacy, Beni-Suef University, Beni-Suef 62514, Egypt. E-mail:Abeer.moawad @ pharm.bsu.edu.eg © 2020 Marwa Elghondakly et al. This is an open access article distributed under the terms of the Creative Commons Attribution 4.0 International License (https://creativecommons.org/licenses/by/4.0/).