stigmasterol and n-hexane

α-Amylase inhibitors from natural sources are of interest for new drug development for the treatment of diabetes mellitus (DM). High-performance thin-layer chromatography (HPTLC) coupled bioassay guided isolation of bioactive compounds has been improved within last few years.. A microchemical derivatised HPTLC-coupled attenuated total reflectance-Fourier-transform infrared (ATR-FTIR) and nuclear magnetic resonance (NMR) spectroscopy was employed for profiling α-amylase inhibitor from the aerial part of Asparagus racemosus Willd.. Asparagus racemosus Willd. aerial part extracted with different solvents (n-hexane, chloroform, ethyl acetate, and methanol) and assayed to detect free radical scavengers and α-amylase inhibitor by 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay and starch-iodine assay method, respectively. HPTLC-coupled ATR-FTIR and NMR spectroscopy was used to identify the α-amylase inhibitor.. Methanolic extract of A. racemosus showed highest antioxidant activity (21.99 μg GAE/μL) where n-hexane extract showed lowest antioxidant activity (5.87 μg GAE/μL). The α-amylase inhibition was recorded as highest and lowest in ethyl acetate extract (13.13 AE/μL) and n-hexane extract (3.92 AE/μL), respectively. The deep blue zone of α-amylase sprayed TLC plate of extracts with hR. The present work establishes the α-amylase inhibiting properties of A. racemosus maintaining its use for the treatment of DM as a traditional medicine. Bioassay guided isolation through HPTLC-coupled ATR-FTIR and NMR spectroscopy offers an effective method for the exploration of bioactive compounds such as α-amylase inhibitor from complex plant extracts.

Topics: Acetates; alpha-Amylases; Antioxidants; Asparagus Plant; Chloroform; Chromatography, Thin Layer; Free Radical Scavengers; Hexanes; Iodine; Magnetic Resonance Spectroscopy; Methanol; Plant Components, Aerial; Plant Extracts; Solvents; Starch; Stigmasterol

Chagas disease is caused by the protozoan parasite Trypanosoma cruzi. This illness is found mainly in 21 Latin American countries and an estimated 8 million people are infected worldwide. The unsatisfactory chemotherapy provokes severe toxicity and resistant strains. Medicinal plants constitute a promising source of new drugs and remedies against all kinds of disorders, mainly infectious diseases arousing interest worldwide.. The aim of this study is the isolation, structural identification and evaluation of the trypanocidal activity of samples present in the Excoecaria lucida Sw. leaves.. Total extract (TE) of E. lucida Sw. leaves was obtained by ethanol extract therefore fractionated sequentially with hexane, ethyl acetate and n-butanol, to obtain three phases: Hex, EA and But, respectively. Ellagic acid (EL1) was purified from both EA and But phases, while EL2; a 1:1 stigmasterol-3-O-β-D-glucopyranoside plus sitosterol-3-O-β-D-glucopyranoside mixture was obtained from the Hex phase. Activity assays were performed using bloodstream and intracellular forms of T. cruzi and cytotoxicity assays using L929 fibroblasts.. The EL1 and EL2 samples were more active against bloodstream trypomastigote forms with EC50 of 53.0±3.6 and 58.2±29.0 µg/mL, respectively; at 100 µg/mL. These samples also showed 70% of inhibition of L929 cells infection. Toxicity assays demonstrated that after 96 h of treatment only the fractions Hex and EA presented detectable cytotoxicity.. Ellagic acid, stigmasterol-3-O-β-D-glucopyranoside and sitosterol-3-O-β-Dglucopyranoside are reported for the first time in E. lucida Sw. leaves as well as their biological activity studies supporting further investigations for Chagas disease treatment.

Topics: 1-Butanol; Acetates; Animals; Ellagic Acid; Euphorbiaceae; Fibroblasts; Glucosides; Hexanes; Mice; Plant Extracts; Plant Leaves; Sitosterols; Stigmasterol; Trypanocidal Agents; Trypanosoma cruzi

Despite several phytochemical studies of Premna serratifolia Linn. (Verbenaceae), the isolation of active constituents of this plant remains to be explored.. The study isolates cytotoxic terpenoids and steroids from the leaves of Premna serratifolia.. Unsaponifiable matter of hexane soluble fraction obtained from methanol extract was subjected to isolation by column chromatography and preparative TLC. Three compounds PS-01 A, PS-01B and PS-02 A were isolated. PS-01 A and PS-01B were identified by comparative TLC with authentic marker compounds followed by NMR analysis. Further PS-01B was analyzed by HR-GCMS. PS-02 A was subjected to HR-LCMS. All isolated compounds/fractions were evaluated for cytotoxic activity by BSL bioassay and using cell lines A549, HepG2 and L6.. Bioactivity guided fractionation of Premna serratifolia leaves succeeded into isolation of two terpenoids and one steroid compound with significant cytotoxic activity. Here we report the isolation of these cytotoxic terpenoids/steroids from this plant for the first time which could be developed as anticancer agents.

Topics: A549 Cells; Animals; Antineoplastic Agents, Phytogenic; Cell Survival; Chromatography, Thin Layer; Dose-Response Relationship, Drug; Gas Chromatography-Mass Spectrometry; Hep G2 Cells; Hexanes; Humans; Inhibitory Concentration 50; Liver Neoplasms; Lung Neoplasms; Magnetic Resonance Spectroscopy; Methanol; Muscle Fibers, Skeletal; Oleanolic Acid; Phytotherapy; Plant Extracts; Plant Leaves; Plants, Medicinal; Rats; Solvents; Stigmasterol; Verbenaceae

We recently reported the characterization of novel cholesterol esterase (EC. 3.1.1.13) from Trichoderma sp. and preliminary work on sterol ester synthesis. In the present study, we further examined the enzyme ability to synthesize cholesterol esters from cholesterol and free fatty acids of various chain lengths, and compared the fatty acid specificity in synthesis with that in hydrolysis. The enzyme catalyzed the synthesis of medium- and long-chain fatty acid cholesterol esters, but failed to synthesize short-chain fatty acid esters. The fatty acid specificities in the synthesis and hydrolysis of cholesterol esters were entirely different from each other. Unlike other lipolytic enzymes, the enzyme was largely independent of water content in the synthesis of cholesterol oleate, and it achieved near-complete esterification in the presence of an equimolar excess of oleic acid. Of additional interest is the finding that the addition of n-hexane markedly enhanced the esterification activities on all the medium- and long-chain saturated fatty acids used. Based on these findings, we attempted to synthesize stigmasterol stearate as a food additive to lower cholesterol levels in blood plasma, and found that the enzyme catalyzed effective synthesis of the ester without the need of dehydration during the reaction, indicating the potential utility of the enzyme in the food industry.

Topics: Anticholesteremic Agents; Biocatalysis; Cholesterol; Cholesterol Esters; Esterification; Fatty Acids; Fatty Acids, Volatile; Food Additives; Food Industry; Fungal Proteins; Hexanes; Hydrolysis; Molecular Structure; Oleic Acid; Oleic Acids; Phytosterols; Stearates; Sterol Esterase; Stigmasterol; Substrate Specificity; Trichoderma; Triolein; Water

We describe a rapid and sensitive method involving time-of-flight secondary-ion mass spectrometry (TOF-SIMS) for specific laboratory diagnosis of the Smith-Lemli-Opitz syndrome, which is characterized by massive (approximately 1000-fold) accumulation of the biosynthetic cholesterol precursor 7-dehydrocholesterol. Minute amounts of blood (1-50 microL) were extracted with n-hexane, and aliquots were analyzed by TOF-SIMS. 7-Dehydrocholesterol and its isomers were detected at 491.3 mass units ([M + 107Ag]+) and cholesterol at 495.3 mass units ([M + 109Ag]+). Quantitation of 7-dehydrocholesterol and cholesterol was achieved after saponification and addition of stigmasterol as internal standard. Whereas 7-dehydrocholesterol and isomeric dehydrocholesterol were not detectable in controls, the patients revealed concentrations ranging between 0.84 and 1.25 mmol/L. Comparison with results obtained by gas chromatography indicated that quantitation by TOF-SIMS yielded the sum of 7-dehydrocholesterol, isomeric dehydrocholesterol II, and sterol III, the latter two also being increased in the patients. Consistent with quantitation by gas chromatography, the cholesterol concentrations in the patients ranged between 1.54 and 2.12 mmol/L (controls: 6.10 +/- 1.37 mmol/L).

Topics: Cholesterol; Chromatography, Gas; Dehydrocholesterols; Hexanes; Humans; Lipid Metabolism, Inborn Errors; Mass Spectrometry; Reference Values; Sensitivity and Specificity; Stigmasterol; Syndrome