621 0009-3130/10/4604-0621 © 2010 Springer Science+Business Media, Inc. Chemistry of Natural Compounds, Vol. 46, No. 4, 2010 NEUTRAL LIPIDS AND BIOLOGICAL ACTIVITY OF THE CHCl 3 EXTRACT OF THE AERIAL PART OF Silene guntensis N. Z. Mamadalieva, 1* N. T. Ulchenko, 1 N. K. Yuldasheva, 1 UDC 547.972:616-006.6 A. A. Zhanibekov, 1 D. R. Egamberdieva, 2 and A. I. Glushenkova 1 Investigations of the plants from the genus Silene showed that they are sources of ecdysteroids [1] and that many of them contain significant quantities of biologically active compounds. We studied lipids occurring in the CHCl 3 extract of S. guntensis B. Fedtsch. during extraction from it of ecdysteroids in order to utilize the plant more completely and safely and to establish the effect of the lipid components on the effectiveness of the antibacterial and cytotoxic activity. Neutral lipids were isolated by separating the extract over a column of silica gel into hydrocarbons (3.34%), esters of fatty acids (FAE) and alcohols (7.83), triacylglycerides (TAG) (2.48), free fatty acids (FFA) (3.12), aliphatic and polyprene alcohols (5.0), and compounds eluted by Et 2 O (19.58), CHCl 3 (3.38), and MeOH (55.26). It can be seen that FAE, FFA, and TAG made up 13.43% of the total. Fatty acids were analyzed as the methyl esters using GLC (Table 1) [2]. Table 1 shows that the FAE and FFA were more enriched in saturated acids than TAG and that the TAG have more unsaturated acids. This is typical for these classes of neutral lipids. The principal saturated acid was palmitic (16:0). Moreover, the FAE fraction included a significant quantity of 24:0 and 26:0 acids (21.3% total) whereas the FFA had 10.3% of the 26:0 acid. The dominant unsaturated acids in these lipid classes were oleic (18:1), linoleic (18:2), and linolenic (18:3). Preliminary screening of the CHCl 3 part of the extract from the aerial part of S. guntensis found that it exhibited antibacterial and cytotoxic activities. Cell Cultivation. Antitumor activity of the tested samples was evaluated using HeLa cells that were cultivated at 37°C under an atmosphere with 90% humidity and 5% CO 2 in 96-well plates (2  10 4 cells/well) in DMEM culture medium (pH 7.4) containing inactivated fetal calf serum (10%), penicillin/streptomycin (100 U/mL), sodium pyruvate (1 mM), and non-essential amino acids (10 mM). After this, the CHCl 3 extract at a concentration of 0.97–500 g/mL was added to the HeLa cells, which were placed in an incubator for 24 h at 37°C. The control was HeLa cells (2  10 4 cells/well) without the extract. The results were calculated in percent of the control. All experiments were conducted in triplicate. MTT-test. Cytotoxic activity was evaluated using the MTT test [3]. Medium in each well was replaced 4 h before the end of cultivation by fresh medium (DMEM, 100 L) containing MTT (0.5 mg/mL). The plates were placed in the incubator for 3 h. The crystals of formazane that were present at the end of the experiment were dissolved in DMSO. Absorption was measured in an apparatus for reading the plates. Cytotoxic activity of the extracts and compounds was determined from the percent reduction in the controls of the color at 595 nm. The preliminary in vitro investigations showed that the CHCl 3 extract of S. guntensis suppressed the growth of HeLa cells. The inhibiting concentration of the CHCl 3 extract according to the MTT tests was a dose of 26.58 g/mL. At that concentration 50% of the cells died compared with the control. This indicated that the extract had antitumor activity. Antibacterial Activity. Cultures of Staphylococcus aureus MRSA16, Micrococcus luteus, Escherichia coli NCTC9001, Pseudomonas aureginosa NCTC6749, Acinetobacter sp. T16, and Staphylococcus saprophyticus T415 were used for the tests. 1) S. Yu. Yunusov Institute of the Chemistry of Plant Substances, Academy of Sciences of the Republic of Uzbekistan, Tashkent, fax: (99871) 120 64 75, e-mail: nmamadalieva@yahoo.com; 2) National University of Uzbekistan, 100174, Uzbekistan. Translated from Khimiya Prirodnykh Soedinenii, No. 4, pp. 523–524, July–August, 2010. Original article submitted February 8, 2010.