Ether Fraction Research Articles

β-amyrin as an analgesic component of the leaves of Callistemon citrinus (Curtis) Skeels: Chemical, biological and in silico studies

The analgesic potential of the leaves of Callistemon citrinus was reported earlier but no active principle for this analgesia was explored. To identify the major analgesic metabolite(s), the leaves were extracted with methanol and fractionated into petroleum ether, carbon tetrachloride, chloroform and aqueous soluble fractions. Based on the thin layer chromatography, the carbon tetrachloride fraction was subjected to gel permeation chromatography and a compound (1) was isolated, which was characterized as β-amyrin. Compound (1) displayed significant peripheral analgesia (p < 0.05) on mice model at an oral dose 200 mg/kg body weight. It also showed noticeable anti-inflammatory membrane stabilizing activity. In silico docking study of β-amyrin with cyclooxygenase (COX)-2 showed a good binding affinity (–9.1 Kcal/mol). Virtual pharmacokinetics and toxicity studies explore its potentials as a lead molecule having no extreme lethality.

Anti-inflammatory and phytochemical evaluation of Combretum aculeatum Vent growing in Sudan

Ethnopharmacological relevanceCombretum aculeatum Vent was traditionally used in Sudan, Eretria and Ethiopia as anti-inflammatory in case of skin inflammation, catarrh, wounds, scorpion stings and snake bites. Nevertheless, there is no scientific information regarding this activity. Aim of studyThe present study aimed to evaluate the phytochemical constituents and the scientific basis for the traditional use of Combretum aculeatum Vent through studying its anti-inflammatory properties for the first time to illustrate the putative mechanisms behind this bioactivity. Materials and methodsthe ethanolic extract was partitioned by petroleum ether, methylene chloride, ethyl acetate, and n-butanol saturated with water. The petroleum ether fraction was saponified and the saponifiable and unsaponifiable fractions were analyzed on GC/MS. The different fractions were subjected to phytochemical investigation to isolate pure compounds. In-vivo anti-inflammatory activity of the ethanolic extract was evaluated using carrageenan induced rat paws edema method at doses of 200, 400 and 600 mg/kg and proved based on histopathological and biochemical parameters. ResultsFive known compounds were isolated for the first time from the aerial parts of Combretum aculeatum Vent: quercetin, vitexin, isorhamnetin 3-O-β-glucoside, isovitexin and rutin, in addition to two previously isolated ones: β-sitosterol and its glucoside. The ethanolic extract evidenced in-vivo anti-inflammatory activity by oral intake of 400 mg/kg of the ethanolic extract significantly (P ≥ 0.05) decreased the paw edema (only 32±1.9% increase in paw weight after 4 h) compared to indomethacin (28.6±2.5%). Moreover, it significantly suppressed the serum malondialdehyde (MDA) and nitric oxide (NO) and increased the GSH to be 11.76±0.85, 5.13±0.62 μmol/mL and 5.66±0.28 μM/mL, respectively. It diminished the serum cytokines TNF-α, IL-6 and IL-1β levels to be 39.1±1.2, 32.6±1.1 and 37.5±1.2 pg/mL, respectively. Results are accompanied by histopathological examination. ConclusionOverall, the results herein presented significant anti-inflammatory properties traditionally ascribed to Combretum aculeatum Vent. Moreover, the biochemical mechanisms associated to this action were highlighted, introducing new prospects for the development of effective anti-inflammatory herbal medicinal products.

Isolation and Characterization of Secondary Metabilites and Evaluation of Antimicrobial, Antioxidant and Thrombolytic Potentials of Erythrina variegata L. Bark

Six compounds, sitosta-4-en-3-one (1), 3β,28-dihydroxyolean-12-en (2), scandenone (3), alpinum isoflavone (4), stigmasterol (5) and lupeol (6) were isolated from the methanol soluble extract of the stem bark of Erythrina variegata. The structure of the compounds was established by extensive NMR studies as well as co-TLC with authentic sample. The petroleum ether, carbon tetrachloride, chloroform and aqueous soluble fractions of the methanolic extract of Erythrina variegata were evaluated for antimicrobial, antioxidant and thrombolytic properties. In the antimicrobial study, most of the fractions of the extract exhibited mild to moderate antimicrobial activity where the zone of inhibition were ranging from 7.69 to 19.51 mm. The chloroform soluble fraction showed significant antioxidant activity with IC50 value 66.28 μg/mL as compared to standard BHT (IC50 value 23.09 μg/mL). The thrombolytic property of different extracts of E. variegata exhibited moderate activity ranging from 31.25 to 57.78 %.

IN VITRO ANTI-INFLAMMATORY ACTIVITY OF METHANOLIC EXTRACT AND DIFFERENT FRACTIONS OF CENTELLA ASIATICA LEAVES BY PROTEIN DENATURATION

Inflammation might be a complex organic reaction to a hazardous stimulant such as pathogens, or injured tissues and mainly causes itching, swelling, skin redness, warm and slight pain. Herbal drugs are widespread in India for their effectiveness, easy availability at low cost and provide low toxicity as compared to modern drugs. Centella asiatica is one of the oldest Ayurvedic medicinal plants, used in treatment of various skin diseases. The aim of our present research was to evaluate the in vitro anti-inflammatory activity of methanolic extract and different fractions of C. asiatica leaves. In protein denaturation method, the percentage inhibition for methanolic extract was observed to be 40.22%. The petroleum ether and n-butanol fraction of methanolic extract of C. asiatica were observed to exhibits 54.12 and 44.42% inhibition, respectively. Diclofenac sodium was used as a standard drug. In comparison with other fractions petroleum ether and n-butanol fractions showed best activity. The preliminary phytochemical studies of n-butanol fractions and n-butanol fractions showed the presence of terpenoids, flavonoids etc., which are used in the treatment of inflammation. Thus, we can call the latter as intense anti-inflammatory agent.

Characterization and hepatoprotective evaluation of sesquiterpenes and diterpenes from the aerial parts of Juniperus sabina L.

Previously, we reported on the hepatoprotective activity of the total extract of Juniperus sabina L. against CCl4 induced liver toxicity in experimental animals. Biologically directed phytochemical study was conducted to identify the active compounds. Male Wistar rats and the standard drug silymarin were used in the study. Hepatoprotective activity was evaluated via serum biochemical parameters such as aspartate amino transferase (AST), alanine aminotransferase (ALT), gamma glutamyl transpeptidase (GGT), alkaline phosphatase (ALP) and total bilirubin. Tissue parameters including non-protein sulfhydryl groups (NP-SH), malonaldehyde (MDA) and total protein (TP) were also determined. Histopathological study was conducted utilizing Mayer’s hematoxylin stain, Periodic Acid Schiff – Hematoxylin (PAS-H) and Masson trichrome technique on light microscope. Electron microscope images were also generated for the study. The activity of the total extract was trapped to the petroleum ether fraction after liquid-liquid fractionation where 51% reduction in the levels of AST, bilirubin and 44% in the levels of ALT were observed. Chromatographic purification of the petroleum ether fraction resulted in the isolation of nine compounds namely: trans-calamenene (1), cadalene (cadalin) (2), epi-cubenol (3), manool (4), calamenene-10β-ol (5), calamenene-10α-ol (6), 4-epi-abietic acid (7), sandaracopimaric acid (8) and isopimaric acid (9). Compounds 1–3, 5 and 6 are belonging to cadinane sesquiterepenes, while compounds 4, 7–9 were of diterpene skeleton. The major compounds were tested for their hepatoprotective effect. Compounds 3 showed marked improvement in the levels of AST and ALT, compound 4 was effective in improving the levels of AST, ALT, GGT, ALP and bilirubin, while compound 7 showed significant improvement in GGT, ALP and bilirubin levels.

Sorbus domestica Leaf Extracts and Their Activity Markers: Antioxidant Potential and Synergy Effects in Scavenging Assays of Multiple Oxidants.

Sorbus domestica leaves are a traditionally used herbal medicine recommended for the treatment of oxidative stress-related diseases. Dry leaf extracts (standardized by LC-MS/MS and LC-PDA) and nine model activity markers (polyphenols), were tested in scavenging assays towards six in vivo-relevant oxidants (O2•−, OH•, NO•, H2O2, ONOO−, HClO). Ascorbic acid (AA) and Trolox (TX) were used as positive standards. The most active extracts were the diethyl ether and ethyl acetate fractions with activities in the range of 3.61–20.03 µmol AA equivalents/mg, depending on the assay. Among the model compounds, flavonoids were especially effective in OH• scavenging, while flavan-3-ols were superior in O2•− quenching. The most active constituents were quercetin, (−)-epicatechin, procyanidins B2 and C1 (3.94–24.16 µmol AA/mg), but considering their content in the extracts, isoquercitrin, (−)-epicatechin and chlorogenic acid were indicated as having the greatest influence on extract activity. The analysis of the synergistic effects between those three compounds in an O2•− scavenging assay demonstrated that the combination of chlorogenic acid and isoquercitrin exerts the greatest influence. The results indicate that the extracts possess a strong and broad spectrum of antioxidant capacity and that their complex composition plays a key role, with various constituents acting complementarily and synergistically.

Gas chromatographic–mass spectrometric profile of non-polar fraction and high-performance thin-layer chromatographic analysis of methanolic fraction with simultaneous quantifications of protocatechuic acid and quercetin in Carissa carandas L. fruits

Extraction of crude drugs by using different solvents provides polarity-based fractions containing specific type of secondary metabolites. Carissa carandas L. fruits were extracted and fractionated, petroleum ether extract was processed by a fatty acid methyl ester (FAME) technique for characterization by gas chromatography—mass spectrometry (GC—MS) analysis, and the remaining part was extracted with methanol for high-performance thin-layer chromatography (HPTLC) analysis, followed by simultaneous quantitative determination of protocatechuic acid and quercetin in methanolic fractions. A validated method for the simultaneous quantification of protocatechuic acid and quercetin was developed and is being reported for the first time in C. carandas L. fruits to the best of our knowledge. Petroleum ether and methanol fractions were found to be the best for the highest possible recovery of targeted analytes. Chromatographic elution of FAME compounds generated from petroleum ether extract was evaluated by GC—MS profiling. Nineteen fatty acid compounds were separated with the highest quantity of octacosanal (13.31%). On the other hand, a polar fraction was processed by HPTLC profiling. For achieving good separation, a mobile phase of toluene-ethyl acetate-formic acid (6:3:1; V/V) was used. Densitometric determination was carried out at 310 nm in the reflection/absorption mode. The calibration curves were linear in the range of 100–600 ng per spot for protocatechuic acid and quercetin. During the analysis, the dried raw material from C. carandas L. fruits showed the presence of protocatechuic acid (0.04%) and quercetin (0.05%). The proposed method is simple, precise, specific, and accurate. The statistical analysis of the data obtained proves that the method is reproducible and selective, which can be used for the routine analysis of the reported phenolic compounds in crude drug and extracts. The simultaneous quantification of these phenolic compounds has not yet been reported in C. carandas L. fruits which may be utilized for the proper standardization of the drug.

INVESTIGATION OF BIOACTIVE CONSTITUENTS AND BIOLOGICAL ACTIVITIES OF DIFFERENT FRACTIONS FROM HERNIARIA HEMISTEMON J.GAY

This study was carried out the phytochemical screening and evaluate antibacterial, antifungal and antioxidants effects of petroleum ether (40-60 °C), diethyl ether, chloroform, ethyl acetate, ethanol 96% and ethanol 70% fractions obtained from the aerial part of Herniaria hemostimon J. Gay. Phytochemical analysis showed the presence of terpenoids, steroids, flavonoids, alkaloids, tannins and saponins in different fractions the plant. Quantitative concentration of phenols, flavonoids, alkaloids, tannins and saponins were detected. HPLC analysis of H. hemostimon identified 21 flavonoids and 21 phenolic compounds. Successive fractionation has been carried out where Di-(2-ethylhexyl) phthalate (DEHP and protocatechuic acid were isolated and identified using FT-IR, 1H-NMR and MS spectroscopic. Furthermore, the antimicrobial activity of successive extract of H. hemistemon against 5 bacterial strains and 2 fungal and yeast strains carried out by the disc diffusion method. The diethyl ether and ethyl acetate fraction showed the highest activity against all the tested bacterial and fungal strains. The ethyl acetate fraction showed the highest antioxidant activity. This study suggested that the bioactivities of H. hemistemona can be used as a source of medicinal compounds due to there's significant antioxidant and antimicrobial activities.

Effect of carpesium abrotanoides petroleum ether fraction on phagocytic function of macrophages and antibody forming cell in mice

Objective To study the effects of carpesium abrotanoides petroleum ether fraction on phagocytic function of macrophages and antibody forming cell in mice. Methods The extract carpesium abrotanoides petroleum ether fraction, forty five mice were divided into the control group and low and high dose groups of carpesium abrotanoides petroleum ether fraction by random number table method, 15 mices in each group. The low-dose and high-dose groups of carpesium abrotanoides petroleum ether fraction were given 50 mg/kg and 100 mg/kg respectively by gavage for 10 days. The spleen weight, growth index, the phagocytosis of macrophages and the number of antibody-forming cells in peritoneal macrophages of mice were detected by chicken erythrocyte phagocytosis test and hemolytic plaque test. Results Compared with the control group, the spleen weight (192.4 ± 11.49 mg, 204.6 ± 10.59 mg vs. 117.6 ± 10.89 mg), the growth index (6.04 ± 0.54, 6.06 ± 0.40 vs. 3.89 ± 0.14), antibody forming (1 216.4 ± 94.1, 1 548.8 ± 86.4 vs.361.0 ± 11.7), phagocytosis percentage of macrophages (58.60% ± 2.60%, 72.0% ± 3.08% vs. 35.49% ± 1.64%), and Phagocytosis index (2.01 ± 0.10, 2.69 ± 0.15 vs. 0.37 ± 0.06) of the groups of low doses and high doses of Carpesium abrotanoides Petroleum ether fraction significantly increased (P<0.05). Conclusions The low doses and high doses of Carpesium abrotanoides Petroleum ether fraction can enhance the mice's specific and nonspecific immune function,and protect the immune system of mice. Key words: Carpesium abrotanoides; Macrophages; Immune function; Mice

Antibacterial, antifungal, and GC–MS studies of Melissa officinalis

Melissa officinalis L. (Lamiaceae) is a perennial plant with varied ethnomedical, therapeutic, and culinary applications. It has been traditionally used in the treatment of various conditions, including cold sores, wounds, infectious bites, ulcers, and parasitic disorders; therefore, this study was undertaken to evaluate the antimicrobial potential of the total ethanol extract and different fractions of M. officinalis against a variety of human pathogenic micro-organisms. Overall, the tested samples demonstrated varying inhibitory potencies against Staphylococcus aureus and Pseudomonas aeruginosa with MICs ranging between 1.65 and 191.40 μg/mL, whereas no antibacterial actions were recorded against Escherichia coli and Klebsiella pneumoniae. The crude extract and its derived fractions also showed noteworthy antifungal activities against Candida albicans, Candida krusei, and Candida glabrata with MICs of 0.30–345.10 μg/mL. Additionally, among the tested plant samples, the petroleum ether fraction exhibited notable antimicrobial spectrum; therefore, it was subjected to GC–MS analysis, for the first time, to ascertain the chemical principles that might underlie its antimicrobial aptitude. Accordingly, a number of metabolites with well-known antimicrobial properties were characterized, mostly dominated by fatty acids and varied terpenoids, with methyl commate A, palmitic acid, and phytol being the major identified components. Interestingly, all the identified constituents are first reported herein from the genus Melissa. The current results collectively highlight the appreciable antimicrobial potential of M. officinalis, particularly against Gram-positive bacteria and yeasts, which strongly endorse its folk use as an anti-infective herb, and reflect its potential as a source of antimicrobial metabolites with possible application both in pharmaceutical and food-related fields.

English

Traditional medicine is widely practiced in Ethiopia. Here we investigate the toxicity of extracts of seven medicinal plants traditionally used to treat breast cancer in Ethiopia. These plants, Sideroxylon oxyacanthum, Zanthoxylum chalybeum, Clematis simensis, Clematis longicauda, Dovyalis abyssinica, Vernonia leopoldi, and Clerodendrum myricoides, were selected based on recommendations by traditional healers and on the frequency of use. After harvesting the plant material, the water content was determined and the powder was subjected to methanol extraction resulting in crude extracts which were tested for cytotoxicity in dose response assay. Then the methanol extract of the most toxic plants was subjected to further solvent-solvent fractionation to gain petroleum ether, hexane, chloroform, ethyl acetate, and water fractions and these were also tested for cytotoxicity in dose response assays. Extracts of Z. chalybeum and C. myricoides were not toxic. The crude extracts of S. oxyacanthum, C. simensis, and D. abyssinica showed cytotoxicity with half maximal inhibitory concentration 50% (IC50) below 1 &mu;g/ml in the human breast cancer cell lines JIMT-1, MCF-7, and HCC1937. The ethyl acetate fraction of V. leopoldi was the most cytotoxic fraction of all fractions tested with an IC50 of 0.87 &mu;g/ml in JIMT-1 cells. The aqueous fraction of S. oxyacanthum and the chloroform fraction of C. simensis were also cytotoxic. In conclusion, our data show a wide difference in in vitro toxicity of medicinal plants used to treat breast cancer patients, which may guide the use of traditional medicine and the choice of plants for isolation of new compounds for cancer treatment. Key words: Cancer, Ethiopia, in vitro cytotoxicity, 3-(4, 5-dimethylthiazolyl-2)-2, 5-diphenyltetrazolium bromide (MTT), traditional medicine. &nbsp

Isolation and Spectral Studies of a New Lactone Triterpenoid from Lantana camara

A new lactone triterpenoid lantanolide along with four known triterpenoids were isolated from the ether fraction of above-ground parts of Lantana camara L. The chemical structure of the new compound was determined to be 3-oxo-12α,22β,25-trihydroxy-18β-olean-28,13β-olide (1), based on spectroscopic data including UV, IR, EI-MS, HR-EI-MS, and 1D and 2D NMR.

Evaluation of anti-ROS and anticancer properties of Tabebuia pallida L. Leaves

BackgroundThe aim of this study was to determine the in-vitro anti-ROS and in-vivo anticancer activities of Tabebuia pallida leaves (abbreviated as TPL) against Ehrlich’s ascite carcinoma (EAC) cells.MethodsThe crude ethanolic extract (CEE) (70% v/v) was fractionated successively with different solvents to get petroleum ether (PEF), chloroform (CHF), ethyl acetate (EAF) and aqueous fraction (AQF). The phytochemical studies were done by standard spectrophotometric methods. The in-vitro cytotoxic and in-vivo anticancer activity were evaluated against brine shrimp nauplii and EAC-induced tumor bearing mice, respectively.ResultsAmong the extracts, the EAF showed the highest phenolic (158.17 ± 1.54 mg/g GAE) and flavonoid (5.43 ± 0.017 mg/g CAE) contents. The total antioxidant capacity of the EAF was 2.30 ± 0.080, which was higher than other extracts and the standard catechin (2.16 ± 0.038). The ferrous reducing antioxidant capacity was in the following order: AA>EAF > CEE > AQF > CHF > PEF. In DPPH and hydroxyl radical scavenging assay, the EAF showed the highest DPPH and hydroxyl radical scavenging activity with IC50 of 6 ± 0.25 μg/ml and 3.58 ± 0.28 μg/ml, respectively, when compared to the standard BHT (IC50 of 16.08 ± 0.28 μg/ml). Also, EAF showed potent inhibitory activity against lipid peroxidation with IC50 of 14.33 ± 0.14 μg/ml. A positive correlation (p < 0.01) between the total phenolics and antioxidant potentials as well as lipid peroxidation inhibition with the hydroxyl radical scavenging was observed. In addition, the EAF showed the moderate cytotoxic effect with ED50of 8.50 ± 0.70 μg/ml and significant cell growth inhibition (71.17 ± 0.44%) when compared to the standards and the cell growth inhibition was positively correlated (p < 0.01) with phenolic contents.ConclusionsOur findings suggest that the leaves of T. pallida might be considered as a new natural source for antioxidant and anticancer compounds, which could be a potent and novel candidate for anticancer therapy.

Antinociceptive activity of petroleum ether fraction obtained from methanolic extract of Clinacanthus nutans leaves involves the activation of opioid receptors and NO-mediated/cGMP-independent pathway

BackgroundMethanol extract (MECN) of Clinacanthus nutans Lindau leaves (family Acanthaceae) demonstrated peripherally and centrally mediated antinociceptive activity via the modulation of opioid/NO-mediated, but cGMP-independent pathway. In the present study, MECN was sequentially partitioned to obtain petroleum ether extract of C. nutans (PECN), which was subjected to antinociceptive study with aims of establishing its antinociceptive potential and determining the role of opioid receptors and l–arginine/nitric oxide/cyclic-guanosine monophosphate (l–arg/NO/cGMP) pathway in the observed antinociceptive activity.MethodsThe antinociceptive potential of orally administered PECN (100, 250, 500 mg/kg) was studied using the abdominal constriction-, hot plate- and formalin-induced paw licking-test in mice (n = 6). The effect of PECN on locomotor activity was also evaluated using the rota rod assay. The role of opioid receptors was determined by pre-challenging 500 mg/kg PECN (p.o.) with antagonist of opioid receptor subtypes, namely β–funaltrexamine (β–FNA; 10 mg/kg; a μ-opioid antagonist), naltrindole (NALT; 1 mg/kg; a δ-opioid antagonist) or nor–binaltorphimine (nor–BNI; 1 mg/kg; a κ-opioid antagonist) followed by subjection to the abdominal constriction test. In addition, the role of l–arg/NO/cGMP pathway was determined by prechallenging 500 mg/kg PECN (p.o.) with l–arg (20 mg/kg; a NO precursor), 1H-[1, 2, 4] oxadiazolo [4,3-a]quinoxalin-1-one (ODQ; 2 mg/kg; a specific soluble guanylyl cyclase inhibitor), or the combinations thereof (l–arg + ODQ) for 5 mins before subjection to the abdominal constriction test. PECN was also subjected to phytoconstituents analyses.ResultsPECN significantly (p < 0.05) inhibited nociceptive effect in all models in a dose-dependent manner. The highest dose of PECN (500 mg/kg) also did not significantly (p > 0.05) affect the locomotor activity of treated mice. The antinociceptive activity of PECN was significantly (p < 0.05) inhibited by all antagonists of μ-, δ-, and κ-opioid receptors. In addition, the antinociceptive activity of PECN was significantly (p < 0.05) reversed by l–arg, but insignificantly (p > 0.05) affected by ODQ. HPLC analysis revealed the presence of at least cinnamic acid in PECN.ConclusionPECN exerted antinocicpetive activity at peripheral and central levels possibly via the activation of non-selective opioid receptors and modulation of the NO-mediated/cGMP-independent pathway partly via the synergistic action of phenolic compounds.

Antidiarrheal, Analgesic, and Anthelmintic Activities of Honeys in the Sundarbans Mangrove Forest, Bangladesh.

This study evaluated the antidiarrheal, analgesic, and anthelmintic activities of honey samples from the Sundarbans mangrove forest of Bangladesh. Composite raw honey (RH), and its diethyl ether (DEH), ethanol (ETH), methanol (MEH), and distilled water (DWH) fractions were investigated. RH and its fractions strongly inhibited castor oil-induced diarrheal episodes in mice at a concentration of 250 mg/kg body weight (b.w.) (P<0.05). At this concentration, RH, DEH, ETH, MEH, and DWH showed inhibitory activity on diarrheal episodes at 43.8, 47.4, 29.8, 12.3, and 38.5%, respectively, whereas for the inhibitory activity for the positive control (PC, 3 mg loperamide/kg b.w.) was 47.4%. Similarly, DEH (250 mg/kg b.w.) showed strongest inhibition (63.5%) of acetic acid-induced writhing in mice, followed by RH (55.7%), ETH (46.2%), MEH (37.6%), and DWH (32.9%). In a hot plate test, mice treated with DEH at a concentration 250 mg/kg b.w. showed the greatest increase in response time, followed by treatment with RH. RH was further used in an anthelmintic test, where it showed a strong dose-dependent reduction in both the paralysis time and the time until death of the parasite, Paramphistomum cervi. Honeys in the Sundarbans could therefore be of great use as nutraceuticals.

Cytotoxicity of Extracts from New Zealand Surf Clams Against Organ Cancer Cell Lines.

In this study, we examined the cytotoxic effects of four fractions from three species of New Zealand (NZ) surf clam on four common organ cancer cells. In most cases, a dose- and time-dependent inhibition on the proliferation of the cancer cells was observed. This was most significant in WiDr (colon) cells, where the percentages of viability reduced to as low as 6%, 5%, and 17% (at 1000 µg 72 h) by extracts from Diamond shell, Storm shell, and Tua tua species, respectively. A549 (lung) cells were the least susceptible to the treatment, with viability percentages at 82%, 15%, and 45%, under the same conditions. Induction of caspase-dependent apoptosis and alterations to the cell cycle further supported the observed morphological analysis. The ethanol, petroleum ether, and ethyl acetate fractions of NZ surf clam, rich in lipids and proteins, were more potent than their water-based counterpart. This is the first demonstration where extracts from NZ surf clams show the ability to inhibit the growth and proliferation of cancer cell lines. We suggest that NZ surf clam extracts have the potential to be further studied and developed as candidates for cancer supplementary management/treatment.

Uji Aktivitas Antioksidan Kombinasi Daun Kopi Arabika (Coffea arabica) dan Daun Pandan (Pandanus amaryllifolius) dengan Metode DPPH

ABSTRACT : Antioxidants are nutrient that can prevent the oxidative damage in the body. Arabica coffee and pandan is most common species of plant found in Indonesia. Arabica coffee and pandan known as plants with antioxidant activity. This study aimed to determine and compare antioxidant activity of water and ether fraction from methanolic extract of Arabica coffee leaves, pandan leaves and both combination by DPPH method. Vitamin C was used as positive control. The result show that Arabica coffee leaves have antioxidant activity better than pandan leaves and also gives big donation of antioxidant activity in their combinations. Statistical analysis showed that antioxidant activity from all sample were significanly different (p<0,05).

Mate Recognition by the Green Mate Borer, Hedypathes betulinus (Coleoptera: Cerambycidae): the Role of Cuticular Compounds

Green mate, Ilex paraguariensis (Aquifoliaceae), is native to South America and its range includes the south of Brazil. Hedypathes betulinus (Coleoptera: Cerambycidae) is considered the most important insect pest of green mate, causing severe damage to plants and significant economic losses. In previous work, laboratory assays were performed to document the mating sequence of this species and the male long-range sex pheromone was identified. Previous reports of the mating sequence suggested that males recognize females only after antennal contact and consequently the existence of a contact sex pheromone. To test the hypothesis that mate-recognition in H. betulinus is mediated by a contact sex pheromone, this study analyzed male and female body extracts by GC-MS. Extracts were fractionated to separate the compounds by polarity, producing hexane, cyclohexene and ether fractions. Active fractions were identified with laboratory bioassays. Bioassays were performed by introducing female cadavers to live males and male copulation response scored (± ). In assays using solvent washed female cadavers, males did not recognize female cadavers, but attempted copulation with female cadavers with the extract re-applied. Cumulatively, these results suggest that a contact sex pheromone on the cuticle of females mediates mate recognition. Laboratory bioassays and analysis of the female solvent extracts produced by fractionation, individually and in binary and ternary combinations, suggests that the more polar compounds present in the ether fraction are crucial to mate recognition in this species.

Evaluation of scavenging activity of hydrogen peroxide in different origins of Liropes Radix by HPLC-UV-CL system

The hydrogen peroxide generation system was used to analyze the scavenging activity of hydrogen peroxide by Liropes Radix from different origins by HPLC-UV-CL. The UV-CL fingerprints of Liropes Radix from different origins were evaluated,and the HPLC-UV and LC-CL fingerprints were systematically analyzed and evaluated. The results showed that the ether fractions of Liriope spicata var. prolifera and L. muscari had good scavenging activity of hydrogen peroxide,and the total activity of different origins varied greatly,while the similar samples had similar activities. The total antioxidant activity of L. muscari is higher than that of L. spicata var.prolifera. The similarity analysis of the two fingerprints was carried out by two different analytical methods. The chemical fingerprints and the active fingerprints have different characteristics. The contribution of each fingerprint to the total peak area and total activity is also different. There are significant differences between the two different fingerprint clustering results.

TLC and GC-MS analysis of petroleum ether fraction of fermented wood "Nikhra" of Acacia seyal

Aim of this study to detect organoleptically Acacia seyal fractions (petroleum ether, chloroform, methanol and aqueous) fermented wood"Nikhra" of the fractions accumulating the strongest sweet fragrance these fragrances were mainly accumulated in the petroleum ether fermented wood "Nikhra" fraction and analysis it by using chromatographic and spectroscopic analysis. Petroleum ether fermented wood "Nikhra" fraction analysis with TLC and spray TLC with vanillin H2SO4 (pink) (C1, C2,C3,C4,C5,C6,C7), Rf values (0.92, 0.86, 0.71, 0.64, 0.57, 0.50, 0.36) ,were expected to be phenolic, with vanillin HCL (red) compounds spots (C5,C6,C7) with Rf values (0.57, 0.50, 0.37) was expected to be catechin and with vanillin H3PO4, blue-violet zones compounds spots (C6, C7), with Rf values (0.50, 0.36), were expected to be lignans. Petroleum ether fermented wood"Nikhra" fraction was divided into two types of compounds classes aromatic and non aromatic by http://research.easybib.com and hence compounds were classified to phenolics and terpenoids compounds by GC/MS. Fragrant aromatics or terpenoids were targeted in this part of study. GC-MS analysis gave a spectrum of fragrance aromatic compounds (phenolics) in the petroleum ether fermented wood "Nikhra" fractions of A.seyal, was Petadecanoic acid(5.64%) and Tetracosamethyl-cyclododecasiloxane(4.17%) total fragrance aromatic compounds(44.57%), and main terponoids compounds was Octadecanoic acid(2.52) % total fragrance aromatic compounds(11.87%).