Formulation,Optimization, and Evaluation of Moringa oleifera Leaf Polyphenol-Loaded Phytosome Delivery System against Breast Cancer Cell Lines

Moringa oleifera leaf polyphenols (Mopp) were encapsulated with phytosomes to enhance their efficacy on 4T1 cancer cell lines. The Mopp were extracted via microwave-assisted extraction. Moringa oleifera polyphenol-loaded phytosomes (MoP) were prepared with the nanoprecipitation method and characterized using the dynamic light scattering and dialysis membrane techniques. The in vitro cytotoxic and antiproliferative activity were investigated with the (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazole) MTT assay. Acute toxicity was assessed using Swiss albino mice. An MoP particle size of 296 ± 0.29 nm, −40.1 ± 1.19 mV zeta potential, and polydispersity index of 0.106 ± 0.002 were obtained. The total phenolic content was 50.81 ± 0.02 mg GAE/g, while encapsulation efficiency was 90.32 ± 0.11%. The drug release profiles demonstrated biphasic and prolonged subsequent sustained release. In vitro assays indicated MoP had a low cytotoxicity effect of 98.84 ± 0.53 μg/mL, doxorubicin was 68.35 ± 3.508, and Mopp was 212.9 ± 1.30 μg/mL. Moreover, MoP exhibited the highest antiproliferative effect on 4T1 cancer cells with an inhibitory concentration of 7.73 ± 2.87 μg/mL and selectivity index > 3. The results indicated a significant difference (p ≤ 0.001) in MoP when compared to Mopp and doxorubicin. The in vivo investigation showed the safety of MoP at a dose below 2000 mg/kg. The present findings suggest that MoP may serve as an effective and promising formulation for breast cancer drug delivery and therapy.     

Chemical Characterization and Nutritional Markers of South African Moringa oleifera Seed Oils

Moringa oleifera Lam (syn. M. ptreygosperma Gaertn.) leaves are globally acclaimed for their nutritional content and mitigation of malnutrition. In most impoverished rural communities including Limpopo, Mpumalanga and KwaZulu Natal of South Africa, powdered leaves of Moringa oleifera are applied as a nutritional supplement for readily available food such as porridge for malnourished children and even breast-feeding mothers. Widely practiced and admired is also the use of the plant seed in the do-it-yourself purification of water by rural South Africans. This study aimed at identifying the chemical and nutritional marker compounds present in South African Moringa oleifera seed oils using high resolution 1-2-dimension gas chromatography in order to give scientific validation to its uses in cosmetics and particularly in culinary practices. Results obtained from two-dimension tandem mass spectrometry chemical signature revealed over 250 compounds, five times more than those reported from one-dimension gas chromatography. Whereas previous reports from gas chromatography-mass spectrometry analysis reported oleic acid (70–78%) as the major compound from oil samples from other countries, M. oleifera seed oil from South Africa is marked by cis-13-octadeaconic acid with 78.62% and 41.9% as the predominant monounsaturated fatty acid in the hexane and dichloromethane extracts respectively. This was followed by cis-vaccenic acid, an isomer of oleic acid at 51% in the acetone extract, 9-octadecanoic acid-(z)-methyl ester at 39.18%, 21.34% and 10.06% in dichloromethane, hexane and acetone extracts respectively. However, a principal component analysis with R² = 0.98 of the two-dimension tandem mass spectrometry cum chemometric analysis indicated n-hexadecanoic acid, oleic acid, 9-octadecanoic acid-(z)-methyl ester and cis-vaccenic acid with a probability of 0.96, 0.88, 0.80 and 0.79 respectively as the marker compounds that should be used for the quality control of moringa seed oils from South Africa. This study demonstrates that South African Moringa oleifera oils contain C-18 monounsaturated fatty acids similar to oils from Egypt (76.2%), Thailand (71.6%) and Pakistan (78.5%) just to mention but a few. These fatty acids are sunflower and olive oil type-compounds and therefore place moringa seed oil for consideration as a cooking oil amongst its other uses.     

Formulation and stability of Moringa oleifera oil microemulsion

Moringa oleifera oil with comparable caprylic triglyceride to Lexol® 865 and higher oleic acid was prepared into microemulsions. A system comprising natural and synthetic oils (O_mix), surfactant (Tween 80 and Span 80; S_mix), and water was microemulsion prepared by a visual grading. Twelve microemulsions with diameter of 83.75–286.76 nm, pH of 6.75–8.23, and viscosity of 280.00–517.20 mPa?s were produced. Their antioxidant were 0.926–2.921 μg Trolox/mg. Microemulsions with O_mix:S_mix mass ratio of 2:8 and 5:5 that had O_mix of 2:1 and 1:1 were stable following a 4 months stability evaluation at 25°C and 45ºC. Microemulsions containing moringa oil produced by a practical low-energy method exhibited good viscosity with an acceptable pH and antioxidant activity were therefore applicable for further applications.     

Isolation and Purification of a Novel Long－chain Acyl Catechin from Lipophilic Tea Polyphenols

Llpophilic tea polyphenols (LTP) was prepared by esterification of green tea polyphenols (GTP) with hexadecanoyl chloride. A novel long-chaln acyl catechin was isolated and purified from LTP by high-speed countercurrent chromatography (HSCCC).Its molecular structure was elucidated as epigallocatechin-3-O-gallate-4‘-O-hexadecanate by elemental analysis, IR, MS and ^1H NMR spectra.     

Proximate,Elemental, and Functional Properties of Novel Solid Dispersions of Moringa oleifera Leaf Powder

Moringa oleifera leaf powder (MOLP) is a rich source of antioxidants, protein, minerals, vitamins, and various phytochemicals and has been used to combat malnutrition in many countries. However, despite its many benefits, MOLP has low a solubility in water, necessitating the development of ways to address this issue. To improve the solubility of MOLP, solid-dispersed Moringa oleifera leaf powders (SDMOLPs) have been developed through freeze-drying, melting, microwave irradiation, and solvent evaporation methods using polyethylene glycols (PEG4000 and PEG6000) (1:1) as hydrophilic carriers. The solid dispersions were evaluated for their proximate composition using standard analytical procedures. Elemental composition was characterized using scanning electron microscopy (SEM) with energy-dispersive X-ray spectroscopy (EDS). Water absorption capacity (WAC) and water-solubility were further evaluated as functional properties. Proximate composition revealed that MOLP and SDMOLPs were rich in protein, energy, carbohydrate, ash, and fat contents. MOLP solid dispersions are a major source of minerals (Ca, Mg, Cu, and Zn), and can be used to alleviate many mineral deficiencies. All solid dispersions had significantly higher (p < 0.05) solubilities (ranging from 54 to 64%) and WAC (ranging from 468.86 to 686.37%), relative to that of pure MOLP. The increased solubility of SDMOLPs may be attributed to the hydrogen bonds and intermolecular interactions between MOLP and the hydrophilic carriers. The results indicate that the solid dispersion technique can be successfully employed to improve the solubility of MOLP. And the solid-dispersed MOLPs with enhanced functional properties may be useful as functional ingredients in foods and beverages, dietary supplements, or nutraceutical formulations.     

Chemical Analysis and Antimicrobial Activity of Moringa oleifera Lam. Leaves and Seeds

Moringa oleifera is a traditional food crop widespread in Asiatic, African, and South American continents. The plant, able to grow in harsh conditions, shows a high nutritional value and medicinal potential evidencing cardioprotective, anti-inflammatory, antioxidant, and antimicrobial properties. The purpose of this study was the phytochemical analysis of M. oleifera and the identification of the antimicrobial compounds by combining a chemical approach with in vitro tests. The metabolite profile of M. oleifera polar and apolar extracts of leaves and seeds were investigated by using Nuclear Magnetic Resonance spectroscopy and Gas Chromatography-Mass Spectrometry. The antimicrobial activity of all of the obtained extract was evaluated against four bacterial pathogens (Staphylococcus aureus, Staphylococcus epidermidis, Pseudomonas aeruginosa and Salmonella enterica). The chemical analysis provided a wide set of metabolites that were identified and quantified. Moreover, apolar extracts from seeds showed a significant concentration-dependent antimicrobial activity against S. aureus and S. epidermidis, (4 mg/mL reduced the viability up to 50%) that was associated to the content of specific fatty acids. Our results remarked the advantages of an integrated approach for the identification of plant metabolites and its use in association with biological tests to recognize the compounds responsible for bioactivity without compounds purification.     

Purification and Characterization of Three Galactose Specific Lectins from Sajna (Moringa Oleifera L.) Leaves

Three lectins designated as SLL‐1, SLL‐2 and SLL‐3 were purified from small sized Sajna (Moringa oleifera L.) leaves by gel filtration of 100% ammonium sulfate saturated crude protein extract on Sephadex G‐75 followed by ion‐exchange chromatography on DEAE and affinity chromatography on Sepharose‐4B. The molecular weight of the lectins SLL‐1, SLL‐2 and SLL‐3 were 1,55,000, 1,15,000 and 85,000, respectively, as determined by gel filtration on Sephadex G‐150 and 1,60,000; 1,20,000 and 85,500, respectively, by SDS‐polyacrylamide slab gel electrophoresis. SLL‐1 and SLL‐2 are dimer in nature held together by disulfide bond (s), while SLL‐3 is a monomer. The lectins agglutinated specifically rat red blood cells and the agglutination was inhibited specifically by methyl‐α‐D‐galactopyranoside, methyl‐β‐D‐galactopyranoside and D‐galactose. The lectins SLL‐1, SLL‐2 and SLL‐3 contain 3.9, 3.4 and 2.8% neutral sugar, respectively, and the sugar compositions were glucose for SLL‐1, mannose for SLL‐2 and SLL‐3 contained either N‐acetyl‐D‐glucosamine or N‐acetyl‐D‐galactosamine or both. The lectins exhibited cytotoxicity in brine shrimp lethality bioassay.     

The Therapeutic Effect and the Potential Mechanism of Flavonoids and Phenolics of Moringa oleifera Lam. Leaves against Hyperuricemia Mice

The aim of this study is to evaluate the anti-hyperuricemia effect and clarify the possible mechanisms of flavonoids and phenolics of MOL (MOL-FP) in mice. Hyperuricemia mice were generated via intraperitoneal (i.p.) administration of potassium oxonate (PO) and oral gavage (p.o.) of hypoxanthine (HX). Serum uric acid (UA), weight, serum XO activity, hepatic XO activity, urea nitrogen (BUN), creatinine (CRE), serum AST level, serum ALT level, mRNA expression of renal urate-anion transporter 1 (URAT1), glucose transporter 9 (GLUT9), organic anion transporters 1 (OAT1), organic anion transporters 3 (OAT3), and ATP-binding cassette transporter G2 (ABCG2) were determined. The molecular docking was conducted using AutoDock Vina 1.2.0 to screen potential XO inhibitors in MOL-FP. Serum metabolomics was established to collect the metabolic profiles of mice and explore the metabolic changes that occurred after MOL-FP treatment. MOL-FP could notably reduce the serum UA level of hyperuricemia mice by inhibiting XO activity and regulating renal urate transporters. Molecular docking studies indicated that 5-p-coumaroylquinic acid, 3-p-coumaroylquinic acid, and catechin could be potential XO inhibitors. Besides, MOL-FP prevented the pathological process of hyperuricemia by regulating biomarkers associated with purine metabolism, amino acid metabolism, and lipid metabolism.     

Effect of Moringa oleifera Leaf Extract on Excision Wound Infections in Rats: Antioxidant,Antimicrobial, and Gene Expression Analysis

The present study investigated the wound healing activity of Moringa oleifera leaf extract on an infected excision wound model in rats. Infection was induced using methicillin-resistant Staphylococcus aureus (MRSA) or Pseudomonas aeruginosa. An investigation was also done to study the effect of Moringa extract on the vascular endothelial growth factor (VEGF) and transforming growth factor-beta 1 (TGF-β1) gene expression in vitro using human keratinocytes (HaCaT). The methanol extract of M. oleifera leaves was analyzed for the presence of phytochemicals by LCMS. The antimicrobial activity of the extract was also determined. Wound contraction, days for epithelization, antioxidant enzyme activities, epidermal height, angiogenesis, and collagen deposition were studied. M. oleifera showed an antimicrobial effect and significantly improved wound contraction, reduced epithelization period, increased antioxidant enzymes activity, and reduced capillary density. Effect of the extract was less in wounds infected with P. aeruginosa when compared to MRSA. The VEGF and TGF-β1 gene expression was increased by M. oleifera.     

Molecular imprinted polymer for solid‐phase extraction of flavonol aglycones from Moringa oleifera extracts

Molecular imprinted polymer produced using quercetin as the imprinting compound was applied for the extraction of flavonol aglycones (quercetin and kaempferol) from Moringa oleifera methanolic extracts obtained using heated reflux extraction method. Identification and quantification of these flavonols in the Moringa extracts was achieved using high performance liquid chromatography with ultra violet detection. Breakthrough volume and retention capacity of molecular imprinted polymer SPE was investigated using a mixture of myricetin, quercetin and kaempferol. The calculated theoretical number of plates was found to be 14, 50 and 8 for myricetin, quercetin and kaempferol, respectively. Calculated adsorption capacities were 2.0, 3.4 and 3.7 μmol/g for myricetin, quercetin and kaempferol, respectively. No myricetin was observed in Moringa methanol extracts. Recoveries of quercetin and kaempferol from Moringa methanol extracts of leaves and flowers ranged from 77 to 85% and 75 to 86%, respectively, demonstrating the feasibility of using the developed molecularly imprinted SPE method for quantitative clean‐up of both of these flavonoids. Using heated reflux extraction combined with molecularly imprinted SPE, quercetin concentrations of 975 ± 58 and 845 ± 32 mg/kg were determined in Moringa leaves and flowers, respectively. However, the concentrations of kaempferol found in leaves and flowers were 2100 ± 176 and 2802 ± 157 mg/kg, respectively.     

In Vitro and In Silico Screening and Characterization of Antimicrobial Napin Bioactive Protein in Brassica juncea and Moringa oleifera

The study aimed to investigate the antibacterial activity of Mustard (Brassica juncea) and Moringa (Moringa oleifera) leaf extracts and coagulant protein for their potential application in water treatment. Bacterial cell aggregation and growth kinetics studies were employed for thirteen bacterial strains with different concentrations of leaf extracts and coagulant protein. Moringa oleifera leaf extract (MOS) and coagulant protein showed cell aggregation against ten bacterial strains, whereas leaf extract alone showed growth inhibition of five bacterial strains for up to 6 h and five bacterial strains for up to 3 h. Brassica juncea leaf extract (BJS) showed growth inhibition for up to 6 h, and three bacterial strains showed inhibition for up to 3 h. The highest inhibition concentration with 2.5 mg/mL was 19 mm, and furthermore, the minimum inhibitory concentration (MIC) (0.5 mg/mL) and MBC (1.5 mg/mL) were determined to have a higher antibacterial effect for <3 KDa peptides. Based on LCMS analysis, napin was identified in both MOS and BJS; furthermore, the mode of action of napin peptide was determined on lipoprotein X complex (LpxC) and four-chained structured binding protein of bacterial type II topoisomerase (4PLB). The docking analysis has exhibited moderate to potent inhibition with a range of dock score −912.9 Kcal/mol. Thus, it possesses antibacterial-coagulant potential bioactive peptides present in the Moringa oleifera purified protein (MOP) and Brassica juncea purified protein (BJP) that could act as an effective antimicrobial agent to replace currently available antibiotics. The result implies that MOP and Brassica juncea purified coagulant (BJP) proteins may perform a wide degree of antibacterial functions against different pathogens.     

A Comprehensive Review with Updated Future Perspectives on the Ethnomedicinal and Pharmacological Aspects of Moringa oleifera

Moringa oleifera is an ancient remedy plant, known as the miraculous plant due to its many prominent uses and significant health benefits. It is a nutrient-rich plant, with exceptional bioactive compounds, such as polyphenols that possess several medicinal properties. Many significant studies have been carried out to evaluate the ethnomedicinal and pharmacological properties of M. oleifera in various applications. Therefore, this comprehensive review compiles and summarizes important findings from recent studies on the potential properties of different parts of M. oleifera. The pharmacological properties of M. oleifera have been studied for various potential biological properties, such as cardio-protective, anti-oxidative, antiviral, antibacterial, anti-diabetic and anti-carcinogenic effects. Therefore, the potential of this plant is even more anticipated. This review also highlights the safety and toxicity effects of M. oleifera treatment at various doses, including in vitro, in vivo and clinical trials from human studies.     

高速逆流色谱分离与鉴定鹿药中黄酮类化合物

采用高速逆流色谱(HSCCC)与其它色谱联用的方法分离纯化鹿药中的化学成分,得到5个黄酮类化合物: 5,7,3′,4′-四羟基-3-甲氧基-8-甲基黄酮(1)、 8-甲基木犀草素(2)、 3′-甲氧基木犀草素(3)、 木犀草素(4)和槲皮素(5),它们均为首次自该种及该属植物中分离得到.HSCCC以V(氯仿)∶ V(甲醇)∶ V(水)=4∶ 3∶ 2混合液为溶剂,上相为固定相,下相为流动相, 分离纯化得到3个分别含化合物1, 4和5的主要部分,经HPLC检验纯度分别为98.3%, 96.7%和95.3%;还有1个含有化合物2和3的较纯部分,通过Sephadex LH-20柱色谱,以V(甲醇)∶ V(水)=1∶ 1混合液洗脱将二者分离.通过理化性质及紫外、红外、质谱、核磁等波谱分析确定化合物结构.     

高速逆流法对刺五加有效成分刺五加苷E的分离制备

 采用高速逆流法分离纯化刺五加粗品中的刺五加苷E。溶剂系统为氯仿 甲醇 异丙醇 水,体积比为5∶6∶1∶4,上相为固定相,下相为流动相,转速为800r/min,流速为2 0mL/min。所得刺五加苷E经高效液相分析,测定纯度为98%(峰面积百分比)。这为其他苷类等极性物质的分离纯化提供了依据。     

Separation and purification of six compounds from the flowers of Cerasus yedoensis (Matsum.) by high-speed countercurrent chromatography in stepwise elution mode

Six compounds from the flower of Cerasus yedoensis (Matsum.) were successfully isolated by high-speed countercurrent chromatography (HSCCC) and preparative high performance liquid chromatography using stepwise elution with a pair of two-phase solvent systems composed of ethyl acetate–n-butanol–formic acid–water at volume ratio of 4:1.5:0.15:5 and ethyl acetate–ethanol–formic acid–water at volume ratio of 4:1:0.15:5 for the first time. This separation process produced (a) 141 mg of 1-O-caffeoyl-β-D-glucopyranoside, (b) 28 mg of p-coumaric acid glucoside, (c) 13 mg of chlorogenic acid, (d) 21 mg of quercetin-3-O-β-D-glucopyranoside, (e) 19 mg of kaempferol 3-O-β-D-glucopyranoside, and (f) 25 mg of caffeic acid from 400 mg of crude sample with the purities of 96.51, 98.82, 94.96, 99.01, 82.51, and 82.45%, respectively. MS, ¹H NMR, and ¹³C NMR analyses were used for the chemical structure identification.     

The Biological and Chemical Ameliorative Effects of Bread Substituted with Dried Moringa Leaves

This study conducted to evaluate the effect of additional dried moringa leaves on bread and their effects on the chemical and biological of rats fed it. Wheat flour with dried moringa leaves powder (DMLB) is used at different levels (6 and 9%) to produce moringa bread. We performed a sensory evaluation of wheat bread substituted with dried moringa leaves, chemical properties (approximate composition and antioxidants), microbiology (total bacteria, spore-forming bacteria, fungi, and yeast) and biological analysis (Triglycerides, total cholesterol, HDL and LDL, ALT, AST, urea, creatinine). Therefore, histopathology examination for kidney and livers of male albino rats fed fortified bread with dried moringa leaves compared with the control sample. The results of the approximate analysis showed significant differences by adding moringa leaves and the percentage of ash, protein, fat, and fiber increased with the moisture content. Moreover, carbohydrates decreased from fortified bread with dried moringa leaves compared to the control sample. The best treatment was fortified bread with 6% dried moringa leaves for all sensory evaluations compared to the other samples and moringa bread (DMLB 9%) helped reduce the microbial load during storage. The effect of moringa bread on liver cirrhosis was evaluated in rats induced by carbon tetrachloride CCl₄. We found out that the effect of moringa leaves on liver and kidney functions in a state of improvement, as for high-density lipoprotein (HDL) and low-density lipoprotein (LDL), the proportion of LDL was less than HDL compared to the control sample. At the same time, moringa leaves have a significant effect on reducing cholesterol, triglycerides, urea, and creatinine in the serum of rats, which may be attributed to the presence of biologically active plant components. It can be concluded that moringa leaves can improve biological and histological of rats feed it.     

芦荟色酮的高速逆流色谱分离制备方法研究

芦荟色酮是在芦荟叶中特有的一类具有抗炎和抑制酪氨酸酶等作用的活性物质。以芦荟全叶为原料,经过一系列的预处理手段,从脱色剂活性炭中获得芦荟色酮粗提物,再经过溶剂分配和富集后采用高速逆流色谱（HSCCC）对其中的色酮成分进行分离纯化。研究结果表明,采用氯仿-甲醇-水(体积比为4∶3∶2)混合溶液和二氯甲烷-甲醇-水(体积比为5∶4∶2)混合溶液作为溶剂分离系统,经过两步HSCCC可以分离纯化出色谱纯度在95％以上的芦荟色酮单体。经过紫外检测、快原子轰击质谱及核磁共振等方法的结构分析鉴定,证实分离所得物质为肉桂酰基-C-葡萄糖甙芦荟色酮。     

Sprouts of Moringa oleifera Lam.: Germination,Polyphenol Content and Antioxidant Activity

(1) Background: In recent years, the consumption of sprouts, thanks to their high nutritional value, and the presence of bioactive compounds with antioxidant, antiviral and antibacterial properties, is becoming an increasingly widespread habit. Moringa oleifera Lam. (Moringa) seems to be an inexhaustible resource considering that many parts may be used as food or in traditional medicine; on the other hand, Moringa sprouts still lack a proper characterization needing further insights to envisage novel uses and applications. (2) Methods: In this study, a rapid and easy protocol to induce the in vivo and in vitro germination of Moringa seeds has been set up to obtain sprouts and cotyledons to be evaluated for their chemical composition. Moreover, the effects of sprouts developmental stage, type of sowing substrate, and gibberellic acid use on the chemical characteristics of extracts have been evaluated. (3) Results: Moringa seeds have a high germinability, both in in vivo and in vitro conditions. In addition, the extracts obtained have different total phenolic content and antioxidant activity. (4) Conclusions: This research provides a first-line evidence to evaluate Moringa sprouts as future novel functional food or as a valuable source of bioactive compounds.