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Effects of ethanol extract of Moringa stenopetala leaves on guinea-pig and mouse smooth muscle.

The fresh leaf ethanol (LE) extract of Moringa stenopetala was tested in guinea-pig ileum and mouse duodenum and in uterus strips. There were significant dose and time dependent reductions of the acetylcholine (AC) response with initial stimulatory effects in both the guinea-pig ileum and the mouse duodenum preparations. Spontaneous rhythmic contractions were greatly reduced, suggesting an antispasmodic property of the crude LE extract. The LE extract showed some oxytocic activity on uterus strips of guinea-pigs and mice. The results are indicative of the traditional use of the leaves of Moringa stenopetala for stomach pain and to expel retained placentae by women.

Animals↗

Hypotensive constituents from the pods of Moringa oleifera.

Hypotensive activity of the ethanolic and aqueous extracts of Moringa oleifera whole pods and their parts, namely, coat, pulp, and seed was investigated. The activity of the ethanolic extract of both the pods and the seeds was equivalent at the dose of 30 mg/kg. The ethyl acetate phase of the ethanolic extract of pods was found to be the most potent fraction at the same dose. Its bioassay-directed fractionation led to the isolation of thiocarbamate and isothiocyanate glycosides which were also the hypotensive principles of the pods as observed in case of Moringa leaves. Two new compounds, O-[2'-hydroxy-3'-(2"-heptenyloxy)]-propyl undecanoate (1) and O-ethyl-4-[(alpha-L-rhamnosyloxy)-benzyl] carbamate (2) along with the known substances methyl p-hydroxybenzoate (3) and beta-sitosterol have also been isolated in the present studies. The latter two compounds and p-hydroxybenzaldehyde showed promising hypotensive activity. Structures of all these compounds have been deduced by spectroscopy and chemical reactions.

Animals↗

Flocculent activity of a recombinant protein from Moringa oleifera Lam. seeds.

Seeds of the tropical tree Moringa oleifera contain small storage proteins able to flocculate particles in suspension in water. The cDNA encoding one of these flocculent proteins, MO(2.1), was cloned and the recombinant protein was expressed in Escherichia coli. The flocculent activity of the purified recombinant MO(2.1)was assayed on clays and bacteria using light and confocal microscopy and GFP-overexpressing bacteria. We show that MO(2.1)is able to aggregate montmorillonite clay particles as well as gram-positive and gram-negative bacteria. We discuss the use of recombinant proteins to study flocculating properties and improve water purification processes.

Amino Acid Sequence↗

Single-step ion exchange purification of the coagulant protein from Moringa oleifera seed.

The coagulant protein from Moringa oleifera (MO) seed was purified using a single-step batch ion exchange (IEX) method. Adsorption and elution parameters were optimized. Impact of the purification on the reduction of organic and nutrient release to the water was studied. The matrix was equilibrated using ammonium acetate buffer, and the optimum ionic strength of NaCl for elution was 0.6 M. The time for adsorption equilibrium was between 90 and 120 min. Maximum adsorption capacity of the matrix, estimated with the Langmuir model, was 68 mg protein/g adsorbent. The purified protein does not release organic and nutrient loads to the water, which are the main concerns of the crude extract. This work suggests that a readily scalable single-step IEX purification method can be used to produce the coagulant protein and it can be carried out with locally available facilities. This will promote the use of MO in large water treatment plants and other industries.

Chromatography, Ion Exchange↗

Enhancing the start-up of a UASB reactor treating domestic wastewater by adding a water extract of Moringa oleifera seeds.

Water extract of Moringa oleifera seeds (WEMOS) was used to enhance the start-up of a self-inoculated upflow anaerobic sludge blanket (UASB) reactor treating raw domestic wastewater. Two reactors labelled control (RC) and WEMOS addition (RM) were started without special inoculum. Both reactors were fed continuously for 22 weeks with domestic wastewater containing an average total chemical oxygen demand (COD) of 320 mg O2/l and suspended solid (SS) of 165 mg/l. The reactors operated during the entire experimental period at 29 degrees C and at a hydraulic retention time (HRT) of 4 h. The RM reactor received 2 ml WEMOS per litre of influent. WEMOS solution was prepared on the basis of 2.5% (w/v) ground M. oleifera seeds in water. The results of 22 weeks' operation showed an improvement in the performance of the RM compared to that of the RC. The dosage of WEMOS in the feed (1) shortened the biological start-up period by 20%, (2) increased acidogenic and methanogenic activity by a factor of 2.4 and 2.2 respectively, (3) increased the specific biogas production by a factor of 1.6, (4) favoured fast growth of the sludge bed, and (5) allowed the aggregation of coccoid bacteria and growth of microbial nuclei, which are precursors of anaerobic granulation.

Anaerobiosis↗

Frying stability of Moringa stenopetala seed oil.

The frying performance of Moringa stenopetala seed oil (extracted with cold press or n-hexane) was studied especially as regards repeated frying operations. The oils were used for intermittent frying of potato slices and cod filets at a temperature of 175 degrees C for 5 consecutive days (5 fryings per day). The chemical changes occurring in oils were evaluated. Free fatty acid content, polar compounds, colour and viscosity of the oils all increased, whereas the iodine value, smoke point, polyunsaturated fatty acid content, induction period and tocopherol content decreased. The effect of the oil on the organoleptic quality of these fried foods and the theoretical number of frying operations possible before having to discard the oil was also determined. The analytical and sensory data showed that the lowest deterioration occurred in cold press produced oil.

Color↗

Anti-fungal activity of crude extracts and essential oil of Moringa oleifera Lam.

Investigations were carried out to evaluate the therapeutic properties of the seeds and leaves of Moringa oleifera Lam as herbal medicines. Ethanol extracts showed anti-fungal activities in vitro against dermatophytes such as Trichophyton rubrum, Trichophyton mentagrophytes, Epidermophyton floccosum, and Microsporum canis. GC-MS analysis of the chemical composition of the essential oil from leaves showed a total of 44 compounds. Isolated extracts could be of use for the future development of anti-skin disease agents.

Antifungal Agents↗

Ternary biosorption studies of Cd(II), Cr(III) and Ni(II) on shelled Moringa oleifera seeds.

Competitive biosorption of Cd(II), Cr(III) and Ni(II) on unmodified shelled Moringa oleifera seeds (SMOS) present in ternary mixture were compared with the single metal solution. The extent of adsorption capacity of the ternary metal ions tested on unmodified SMOS was low (10-20%) as compared to single metal ions. SMOS removed the target metal ions in the selectivity order of Cd(II) > Cr(III) > Ni(II). Sorption equilibria, calculated from adsorption data, explained favorable performance of biosorption system. Regeneration of exhausted biomass was also attempted for several cycles with a view to restore the sorbent to its original state.

Adsorption↗

High-performance liquid chromatography method to measure alpha- and gamma-tocopherol in leaves, flowers and fresh beans from Moringa oleifera.

A high-performance liquid chromatography method for the microscale determination of alpha- and gamma-tocopherol in leaves, flowers and fresh beans from Moringa oleifera is reported. The method includes microscale saponification and extraction with n-hexane. Optimized conditions for reversed-phase HPLC with UV detection were as follows: column, 25 cm x 0.46 cm, Exil ODS 5-microm; column temperature, 25 degrees C; mobile phase, a 20:80 (v/v) mixture of methanol:acetonitrile; flow rate, 1.0 ml/min. With these conditions, method precision (relative standard deviation) was 5.6% for alpha-tocopherol and 4.9% for gamma-tocopherol. We used this method to measure alpha- and gamma-tocopherol in samples from M. oleifera as part of nutritional studies in edible plants cultivated in the Northwest México.

Chromatography, High Pressure Liquid↗

Interfacial properties and fluorescence of a coagulating protein extracted from Moringa oleifera seeds and its interaction with sodium dodecyl sulphate.

The surfactant behaviour of aqueous coagulating protein extracted from Moringa oleifera seeds has been investigated by surface tension measurements. The interaction of the coagulant protein with an anionic surfactant sodium dodecyl sulphate (SDS) has been monitored by surface tension and intrinsic protein fluorescence measurements. The extracted protein shows some weak surface activity at low concentrations. To achieve maximum surface activity (i.e. maximum reduction in surface tension of water), substantially higher concentrations of protein are required. The coagulant protein-SDS interaction scheme did not exhibit the behaviour of weakly interacting polymer-surfactant systems and the SDS interacts in a monomeric form with the protein. The association process of SDS with the coagulant protein is supported by protein fluorescence measurements. SDS has an effect on the fluorescence of the coagulant protein indicating that the local environment of tryptophan in the protein changes as SDS concentration below its critical micelle concentration is increased. These results have led us to the conclusions that: (1) the protein extracted from M. oleifera seeds has significant surfactant behaviour; (2) the coagulant protein interacts strongly with SDS and the protein might have specific binding sites for SDS; (3) there is formation of protein-SDS complex.

Fluorescence↗

Sorption potential of Moringa oleifera pods for the removal of organic pollutants from aqueous solutions.

Moringa oleifera pods Lamarck (Drumstick or Horseradish) is a multipurpose medium or small size tree from sub-Himalayan regions of north-west India and indigenous to many parts of Asia, Africa, South America, and in the Pacific and Caribbean Islands. Its pods (MOP) have been employed as an inexpensive and effective sorbent for the removal of organics, i.e., benzene, toluene, ethylbenzene and cumene (BTEC) from aqueous solutions using HPLC method. Effect of different parameters, i.e., sorbent dose 0.05-0.8g, 25cm(-3) agitation time 5-120min, pH 1-10, temperature 283-308K and concentration of sorbate (1.3-13)x10(-3), (1.1-11)x10(-3), (0.9-9)x10(-3), (0.8-8)x10(-3)moldm(-3), on the sorption potential of MOP for BTEC have been investigated. The pore area and average pore diameter of the MOP by BET method using nitrogen as a standard are calculated to be 28.06+/-0.8m(2)g(-1) and 86.2+/-1.3nm respectively. Freundlich, Langumir and Dubinin-Radushkevich (D-R) sorption isotherms were employed to evaluate the sorption capacity of MOP. Sorption capacities of BTEC onto MOP have been found to be 46+/-10, 84+/-9, 101+/-4, 106+/-32mmolg(-1) by Freundlich, 8+/-0.1, 9+/-0.1, 10+/-0.3, 9+/-0.1mmolg(-1) by Langumir and 15+/-1, 21+/-1, 23+/-2, 22+/-3mmolg(-1) by D-R isotherms respectively, from BTEC solutions at 303K. While the mean energy of sorption process 9.6+/-0.3, 9.2+/-0.2, 9.3+/-0.3, 9.5+/-0.4kJmol(-1) for BTEC is calculated by D-R isotherm only. Rate constant of BTEC onto MOP 0.033+/-0.003, 0.030+/-0.002, 0.029+/-0.002, 0.027+/-0.002min(-1) at solution concentration of 1.3x10(-3), 1.1x10(-3), 0.9x10(-3) and 0.8x10(-3)moldm(-3) and at 303K have been calculated by employing Lagergren equation. Thermodynamic parameters DeltaH -8+/-0.4, -10+/-0.6, -11+/-0.7, -11+/-0.7kJmol(-1), DeltaS -22+/-2, -26+/-2, -27+/-2, -29+/-3Jmol(-1)K(-1) and DeltaG(303K) -0.9+/-0.2, -1.9+/-0.2, -2.3+/-0.1 and -2.6+/-0.2kJmol(-1) were also estimated for BTEC respectively at temperatures 283-308K. The negative values of DeltaH, DeltaS and DeltaG suggest exothermic, stable (with no structural changes at solid-liquid interface) and spontaneous nature of sorption process under optimized conditions. MOP has been used extensively to accrue and then to preconcentrate benzene, toluene and ethylbenzene in wastewater sample.

Adsorption↗

Innovative physico-chemical treatment of wastewater incorporating Moringa oleifera seed coagulant.

Moringa oleifera is a pan tropical, multipurpose tree whose seeds contain a high quality edible oil (up to 40% by weight) and water soluble proteins that act as effective coagulants for water and wastewater treatment. The use of this natural coagulant material has not yet realised its potential. A water extract of M. oleifera seed was applied to a wastewater treatment sequence comprising coagulation-flocculation-sedimentation-sand filtration. The study was laboratory based using an actual wastewater. Overall COD removals of 50% were achieved at both 50 and 100mg/l M. oleifera doses. When 50 and 100mg/l seed doses were applied in combination with 10mg/l of alum, COD removal increased to 58 and 64%, respectively. The majority of COD removal occurred during the filtration process. In the tests incorporating alum, sludge generation and filter head loss increased by factors of 3 and 2, respectively. These encouraging treatment results indicate that this may be the first treatment application that can move to large scale adoption. The simple water extract may be obtained at minimal cost from the presscake residue remaining after oil extraction from the seed. The regulatory compliance issues of adopting 'new materials' for wastewater treatment are significantly less stringent than those applying to the production of potable water.

Environmental Restoration and Remediation↗

Pretreatment of palm oil mill effluent (POME) using Moringa oleifera seeds as natural coagulant.

Moringa oleifera seeds, an environmental friendly and natural coagulant are reported for the pretreatment of palm oil mill effluent (POME). In coagulation-flocculation process, the M. oleifera seeds after oil extraction (MOAE) are an effective coagulant with the removal of 95% suspended solids and 52.2% reduction in the chemical oxygen demand (COD). The combination of MOAE with flocculant (NALCO 7751), the suspended solids removal increased to 99.3% and COD reduction was 52.5%. The coagulation-flocculation process at the temperature of 30 degrees C resulted in better suspended solids removal and COD reduction compared to the temperature of 40, 55 and 70 degrees C. The MOAE combined with flocculant (NALCO 7751) reduced the sludge volume index (SVI) to 210mL/g with higher recovery of dry mass of sludge (87.25%) and water (50.3%).

Flocculation↗

Detection of water soluble lectin and antioxidant component from Moringa oleifera seeds.

Seed flour from Moringa oleifera is widely used as a natural coagulant for water treatment in developing countries. Extracts obtained by water soaking of M. oleifera intact seeds were investigated for the presence of lectin, trypsin inhibitor, tannin as well as antioxidant activity. A water soluble M. oleifera lectin (WSMoL) detected was mainly active with rabbit cells at pH 4.5; heat treatment, pH 7.0, fructose and porcine thyroglobulin abolished HA of WSMoL. Trypsin inhibitor or tannins were not detected; the antioxidant component (WSMoAC) reduced 1,1-diphenyl-2-picrylhydrazyl radical (DPPH) was slower than catechin and was thermostable. The extracts showed a primary glycopolypeptide band of Mw 20,000; the main native acidic protein showed hemagglutinating activity. WSMoL may be involved in seed coagulant properties.

Animals↗

A simple purification and activity assay of the coagulant protein from Moringa oleifera seed.

Use of extracts from Moringa oleifera (MO) is of great interest for low-cost water treatment. This paper discusses water and salt extraction of a coagulant protein from the seed, purification using ion exchange, its chemical characteristics, coagulation and antimicrobial properties. The coagulant from both extracts is a cationic protein with pI greater than 9.6 and molecular mass less than 6.5 kDa. Mass spectrometric analysis of the purified water extract indicated that it contained at least four homologous proteins, based on MS/MS peptide sequence data. The protein is thermoresistant and remained active after 5h heat treatment at 95 degrees C. The coagulant protein showed both flocculating and antibacterial effects of 1.1--4 log reduction. With samples of high turbidity, the MO extract showed similar coagulation activity as alum. Cecropin A and MO extract were found to have similar flocculation effects for clay and microorganisms. Simple methods for both the purification and assay of MO coagulating proteins are presented, which are necessary for large-scale water treatment applications.

Anti-Bacterial Agents↗

(13)C NMR characterization of triacylglycerols of Moringa oleifera seed oil: an "oleic-vaccenic acid" oil.

The composition of acyl chains and their positions in the triacylglycerols of the oil extracted from seeds of Moringa oleifera were studied by (13)C NMR spectroscopy. The unsaturated chains of M. oleifera seed oil were found to comprise only mono-unsaturated fatty acids and, in particular, two omega-9 mono-unsaturated acids, (cis-9-octadecenoic (oleic acid) and cis-11-eicosenoic acids) and one omega-7 mono-unsaturated acid (cis-11-octadecenoic acid (vaccenic acid)). The mono-unsaturated fatty acids were detected as separated resonances in the spectral regions where the carbonyl and olefinic carbons resonate according to the 1,3- and 2-positions on the glycerol backbone. The unambiguous detection of vaccenic acid was also achieved through the resonance of the omega-3 carbon. The (13)C NMR methodology enabled the simultaneous detection of oleate, vaccenate, and eicosenoate chains according to their positions on the glycerol backbone (1,3- and 2-positions) through the carboxyl, olefinic, and methylene envelope carbons of the triacylglycerol acyl chains.

Carbon Isotopes↗

Glucosinolates in Moringa stenopetala.

Bio-guided fractionation of seed extracts from Moringa stenopetala resulted in a myrosinase hydrolysis product, 5,5-dimethyloxazolidine-2-thione. It is formed from the glucosinolate glucoconringiin, which was identified together with O-(rhamnopyranosyloxy)benzyl glucosinolate from M. stenopetala for the first time. The glucosinolates in seeds, leaves and roots of M. stenopetala were quantified as des-sulphoglucosinolate by HPLC. The seeds without testa contained the highest concentration of glucoconringiin and of O-(rhamnopyranosyloxy)benzyl glucosinolate within the plant, 3 % and 19 % of dry mass, respectively. Abbreviations. GLS:glucosinolate GC:glucoconringiin RB-GLS: O-(rhamnopyranosyloxy)benzyl glucosinolate

Chromatography, High Pressure Liquid↗

Effects of Moringa oleifera seed extract on rumen fermentation in vitro.

Moringa oleifera is a pantropical tree of the family Moringaceae. A previously undescribed property of an aqueous extract from the seeds of this plant is the modulation of ruminal fermentation patterns, especially protein degradation, as demonstrated in a short-term batch incubation system. Gas, short chain fatty acids (SCFA) and cellulolytic enzyme activities were determined as general fermentation parameters. A dot blot assay able to directly detect true protein in rumen fluid samples was used to quantify protein degradation. For complex substrates the interpretation of protein degradation profiles was amended by polyacrylamide gel electrophoresis (PAGE) of the samples. When incubated with pure carbohydrates at a concentration of 1 mg ml(-1), the extract reduced microbial degradation of the model protein, bovine serum albumin (BSA), such that its concentration was at least 40% above the control after 12 h of incubation. Total protein degradation was thus delayed by approximately 9 h. When fermented along with wheat straw, leaf protein (Rubisco) was almost entirely protected during 12 h of fermentation. The degradation of soy proteins was retarded by at least 4-6 h, depending on the protein band. There were strong side effects on the fermentation of pure cellulose (SCFA yield-60% after 12 h), whereas cellobiose and starch fermentation were less affected (-18 and -8%, respectively). When the complex substrates were fermented, SCFA yield was reduced by approximately 30% after 12 h. In our work we clearly demonstrate the efficacy of the new substance, which is neither a tannin nor a saponin, in an in vitro system, using pure as well as complex substrates. The properties shown in vitro for the crude extract suggest that it could have a positive effect on the protein metabolism of ruminants under intensive management and that negative side effects can be overcome by an optimized dosage. If the chemical nature of the active substance and its mechanism of action can be clarified, it may provide an alternative to replace critical synthetic feed additives (such as antibiotics) for high yielding dairy cows.

Animals↗