Shahira M. Ezzat

Isolation of biologically active constituents from Moringa peregrina (Forssk.) Fiori. (family: Moringaceae) growing in Egypt

Background: Moringa peregrina is a wild plant that grown in the eastern desert mountains in Egypt. Although, this plant is native to Egypt, no details studies were traced on its chemical composition and biological activity. Materials and Methods: The different fractions of the ethanolic extract of the dried aerial parts of the plants were subjected to fractionation and purification on various silica and sephadex columns for the isolation of the major compounds which were tested for there anticancer activity. The aqueous and ethanolic extract as well as its different fractions were tested for antihyperglycemic effect on Streptozitocin-induced diabetes in rats. Results: Investigation of the different fractions of the ethanolic extract of the aerial parts of M. peregrina yielded lupeol acetate (1), β-amyrin (2), α-amyrin (3), β-sitosterol (4), β-sitosterol-3-O-glucoside (5), apigenin (6), rhamnetin (7), neochlorogenic acid (10), rhamnetin-3-O-rutinoside (12), and 6-methoxy-acacetin-8-C-β-glucoside (13) which were isolated for the first time from the plant. Compound (13) was isolated for the first time from genus Moringa. In addition, quercetin (8), chryseriol-7-O-rhamnoside (9) and quercetin-3-O-rutinoside (11) were also isolated. Identification has been established by spectral data (UV, MS, IR, 1H, 1H -1H COSY, and 13C-NMR). The major isolated compounds were found to have valuable cytotoxic activities against breast (MCF 7) and colon (HCT 116) cancer cell lines and their activities were comparable to the reference drug doxorubicin. On the other hand, the aqueous and ethanolic extracts as well as the n-hexane fraction were found to have potent antihyperglycemic effect on Streptozitocin-induced diabetes in rats. Conclusion: The Egyptian plant M. peregrina is rich in biologically active ingredients which showed potent cytotoxic activity and also its ethanolic extraxt exert a significant antihyperglycemic effect.

Chemical and biological study of the seeds of Eragrostis tef (Zucc.) Trotter

A chemical study of the red type of the seeds of Eragrostis tef (Zucc.) Trotter (Poaceae) led to the isolation of seven compounds from its ethanol extract, namely β-sitosterol (1), β-amyrin-3-O-(2′-acetyl(glucoside (2), β-sitosterol-3-O-β-D-glucoside (3), naringenin (4), naringenin-4′-methoxy-7-O-α-L-rhamnoside (5), eriodictyol-3′,7-dimethoxy-4′-O-β-D-glucoside (6) and isorhamnetin-3-O-rhamnoglucoside (7), which are reported for the first time in the genus Eragrostis. This is the first report for the isolation of compounds (2) and (6) in nature. The isolated compounds were identified using different spectroscopic methods (EIMS, UV, 1H NMR, 13C NMR, HMQC and HMBC). A proximate analysis revealed the high nutritive value of the seeds: carbohydrates (57.27%), protein (20.9%), essential amino acids (8.15%) with major leucine and lysine (1.71 and 1.35%, respectively), vitamin B1 (1.56u2009mg/100u2009g) and potassium and calcium (32.4 and 9.63%, respectively). The seeds yielded 22% w/w of fixed oil rich in unsaturated fatty acids (72.46%), among which oleic acid, was predominant (32.41%), followed by linolenic acid (23.83%). The ethanolic extract and fixed oil of the seeds exhibited anti-hyperlipedaemic and anti-hyperglycaemic activities. Oral administration of the fixed oil for 10 days resulted in a rise in serum calcium levels in rats.

Effect of the Method of Preparation on the Composition and Cytotoxic Activity of the Essential Oil of Pituranthos tortuosus

The aerial parts of Pituranthos tortuosus (Desf.) Benth and Hook (Apiaceae), growing wild in Egypt, yielded 0.8%, 0.6%, and 1.5% (v/w) of essential oil when prepared by hydrodistillation (HD), simultaneous hydrodistillation-solvent (n-pentane) extraction (Lickens- Nickerson, DE), and conventional volatile solvent extraction (preparation of the “absolute”, SE), respectively. GC-MS analysis showed that the major components in the HD sample were β-myrcene (18.81%), sabinene (18.49%), trans-iso-elemicin (12.90%), and terpinen- 4-ol (8.09%); those predominent in the DE sample were terpinen-4-ol (29.65%), sabinene (7.38%), γ-terpinene (7.27%), and β-myrcene (5.53%); while the prominent ones in the SE sample were terpinen-4-ol (15.40%), dill apiol (7.90%), and allo-ocimene (4E,6Z) (6.00%). The oil prepared in each case was tested for its cytotoxic activity on three human cancer cell lines, i.e. liver cancer cell line (HEPG2), colon cancer cell line (HCT116), and breast cancer cell line (MCF7). The DE sample showed the most potent activity against the three human cancer cell lines (with IC50 values of 1.67, 1.34, and 3.38 μg/ml against the liver, colon, and breast cancer cell lines, respectively). Terpinen-4-ol, sabinene, γ-terpinene, and β-myrcene were isolated from the DE sample and subjected to a similar evaluation of cytotoxic potency; signifi cant activity was observed

Determination of bioactive markers in Cleome droserifolia using cell-based bioassays for antidiabetic activity and isolation of two novel active compounds

The antidiabetic activities of the aqueous (AqEx) and ethanolic (AlEx) extracts of Cleome droserifolia (Forssk.) Del., were tested in cultured C2C12 skeletal muscle cells and 3T3-L1 adipocytes. An 18-h treatment with the AqEx increased basal glucose uptake by 33% [insulin equivalent (IE)=1.3±0.04] in muscle cells comparable to a 25.5% increase caused by 100 nM insulin (IE=1±0.03). Fractionation of the tested AqEx yielded hexane (HxFr), chloroform (ClFr) and ethyl acetate (EtFr) fractions which exerted 38, 52 and 35% increase in the glucose uptake corresponding to an IE of 1.5±0.06, 2.0±0.04 and 1.4±0.04, respectively. Only the ClFr and EtFr accelerated the triglyceride accumulation [rosiglitazone equivalent (RE) was 0.9±0.13 and 0.63±0.12, respectively] in pre-adipocytes undergoing differentiation comparably with 10 μM rosiglitazone. Six terpenoids (C1-C6) and three flavonol glycosides (F1-F3) were isolated from the active ClFr and EtFr, respectively, and identified. C5, C2 and C4 had an IE of 0.86±0.05, 1.01±0.04 and 0.9±0.08, while F1, F2 and F3 gave an IE of 1.3±0.05, 2.3±0.05 and 2.0±0.04, respectively. We could conclude that the reported antihyperglycemic activity of Cleome droserifolia is attributed to significant insulin-like effects in peripheral tissues, and that compounds F2 and F3, being highly active, could be used as bioactive markers to standardize the C. droserifolia herbal extract.

Phytochemical and biological investigation of the extracts of Nigella sativa L. seed waste.

Different extracts of Nigella sativa L. seed waste; aqueous (AE) 200 mg/kg, ethanol 70% (EE) 250 mg/kg and hexane (HE) 10 mg/kg, were evaluated for their hepatoprotective activities. They were administered orally, once daily, for 5 consecutive days. On day 5, liver injury was induced in animals by a single i.p. injection of carbon tetrachloride (10 mg/kg b. w. of 0.25% (v/v). Hepatoxicity produced, was evaluated by both biochemical and histopathological investigations. The aqueous extract attenuated the CCl(4) -induced liver damage likely due to the decrease of proinflammatory cytokines and T-cell proliferation. This was noticed by a significant decrease in both serum and tissue cytokines; tumor necrosis factor-alpha (TNF-α), interferon-gamma (INF-γ) and interlukin-beta (IL-1β), in the markers of liver functions; bilirubin and glutamic pyruvic transaminase (GPT) and in the oxidative stress markers; malondialdehyde (MDA) and glutathione content (GSH). Fractionation of this extract was performed and its component, protein, saponin, and polyphenol fractions were evaluated by appropriate analytical procedures. The crude protein of the seed waste reached 36.85% while protein fingerprint showed four bands ranging from 91.97 KD and 29.00 KD. The saponin content was evaluated through the determination of the haemolytic index and reached 15.56 mg/g dry powder. Finally, Folin Ciocalteu method was used for the determination of the total polyphenols. The same biochemical and histopathological studies were again performed on the different fractions of the aqueous extract; protein fraction (PF) 10 mg/kg, saponin fraction (SF) 5 mg/kg and polyphenol fraction (FF) 10 mg/kg. The biochemical changes were improved only by the protein fraction (PF) of the seed waste of Nigella sativa. This was manifested by a significant reduction in both serum and tissue cytokines in the liver markers and in the oxidative stress markers. Moreover, liver histopathology showed that (PF) reduced the incidence of liver lesions including hepatic cells cloudy swelling, lymphocytes infiltration, hepatic necrosis and fibrous connective tissue proliferation induced by CCl(4) in mice. From this study, it is concluded that the protein fraction of the aqueous extract of Nigella sativa seed waste exhibited a promising hepatoprotective effect in the management of different liver disorders.

Chemical and biological investigation of Araucaria heterophylla Salisb. resin.

Three labdane diterpenes, namely labda-8(17),14-diene, 13-epicupressic acid, and 13-Oacetyl- 13-epicupressic acid, were isolated from the resin collected from stem exudates of Araucaria heterophylla Salisb. (Araucariaceae). The isolated compounds were identified using different spectroscopic methods (1H NMR, 13C NMR, HMQC, HMBC and COSY). The resin extract showed antiulcerogenic activity against ethanol-induced stomach ulcers in Sprauge Dawely rats using ranitidine as standard. In addition, the resin and the isolated compounds showed variable cytotoxic activities against breast (MCF7) and colon (HCT116) cancer cell lines.

Hepatoprotective constituents of Torilis radiata Moench (Apiaceae)

An investigation of the aqueous ethanolic extract (AE) of the aerial parts of Torilis radiata Moench yielded two triterpenes (lupeol acetate (1) and α-amyrin (2)), a sterol (spinasterol (3)) from its n-hexane fraction (HF), a flavone (acacetin (4)), a coumarin (scopoletin (5)), a phenolic acid (ferulic acid (6)) from the chloroform fraction (CF) and a flavone glycoside (luteolin-7-O-glucoside (7)) from the n-butanol fraction (BF). The hepatoprotection of the AE and its fractions was assessed in terms of the reduction in histological damage, accompanied by restoration of the liver enzymes (alanine amino transferase (ALT), aspartate amino transferase (AST), lactate dehydrogenase (LDH)), a reduction in the inflammatory markers (tumour necrosis-α (TNF-α), nitric oxide (NO), N-acetyl-β-D-glucosaminidase (NAG) and myloperoxidase (MPO) in serum) and restoration of the oxidant balance by decreasing the serum and hepatic malondialdehyde (MDA) levels, along with increasing the activity of hepatic catalase (CAT), glutathione peroxidase (GSHPx) and the non-enzymatic antioxidant glutathione (GSH).

Chemical and biological study of Mentha suaveolens Ehrh. cultivated in Egypt

Investigation of the different fractions of the ethanolic extract of the aerial parts of M. suaveolens growing in Egypt yielded nine compounds; two new triterpenes [3β-acetyl -22α-hydroxy ursa-12,20diene (compound 1) and 2α, 3β-dihydroxy-olean-18-en-29-oic acid (compound 7)] and nine known compounds: a sterol and its glucoside; β-sitosterol (compound 2) and β-sitosterol-3-O- β-D-glucoside (compound 4), a triterpene; oleanolic acid (compound 3), two monoterepenes; dihydrolimonene (compound 5) and 7-hydroxy-p-cymene (compound 6), two flavonoids; isoquercitrin (compound 8) and rutin (compound 9) which were isolated from M. suaveolens Ehrh. for the first time. The structures of the isolated compounds were identified by spectral data (UV, MS, 1D and 2D-NMR) and comparison with authentic samples. Moreover, the ethanolic extract showed potent analgesic activity as compared to indomethacin. The ethyl acetate fraction was the most potent as anti-inflammatory (88%), followed by the ethanolic extract (82.9%) as compared with indomethacin. The ethanolic extract and its four subfractions showed a moderate inhibitory activity against the tested human pathogenic bacteria.

Effect of Certain Essential oils on Dissolution of Three Commercial Gutta-percha Brands

Abstract The purpose of this study was to test the ability of some essential oils to dissolve three types of commercial gutta-percha brands which are commonly used in Egypt. The chemical composition of three commercially available gutta-percha brands (Hygenic, Maillefer and Meta) was determined using X-ray, FTIR and elemental microanalysis. Essential oils from thirty medicinal plants representing the six most famous families of aromatic plants growing in Egypt were extracted by steam distillation and tested for dissolving the gutta-percha brands. Essential oils from the peels of Citrus x sinensis (L.) Osbeck (Navel orange), C. deliciosa Tenora (Common mandarin) and C. x aurantium L.(Bitter orange)(Rutaceae) showed the highest dissolution rate. The essential oils from the aerial parts of Origanum syriacum L. and Lavandula hyprida Reverchon cv. Grosso (Labiateae) showed complete dissolution at 40oC, also essential oils from the seeds of Elletaria cardamomum (Cardamom) Matonvar. miniscula Burkill (Zingibraceae) showed similar dissolution at 40oC. GC-MS analysis was performed on six essential oils representing the three effective families in order to correlate their composition with their dissolution efficiency.