Phyto-constituents profiling of Luffa echinata and in vitro assessment of antioxidant,anti-diabetic,anticancer and anti-acetylcholine esterase activities

Luffa echinata Roxb. is one of the neglected medicinal plants. It is an important source of bioactive metabolites and used in several Ayurvedic formulations. In the present analysis, mature leaves and fruits were extracted with acetone, ethanol, acetonitrile, methanol and water. Phytochemicals like total phenolic (TPC), flavonoid (TFC), tannin (TTC), alkaloid (TAC) and terpenoid (TTEC) content were analysed. Further, antioxidant (AOX) activities like 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity, 2,2′-azino-bis-(3-ethyl) benzothiazoline-6-sulfonic acid (ABTS) radical scavenging, metal chelating activity (MC), ferric reducing antioxidant power (FRAP) and phosphomolybdenum assay (PMA) were studied. Highest TPC and TFC (189.57 ± 1.9 mg TAE/g extract, 30.48 ± 0.7 mg CE/g extract, respectively) were reported from acetone extract of the leaves. Ethanolic fruit extract showed the highest TTC (13.79 ± 0.2 mg CE/g extract). Acetone and acetonitrile fruit extract revealed maximum TTEC (602.79 ± 3.5 mg UAE/g extract) and moderate TAC (19.96 ± 0.9 mg GE/g extract), respectively. In AOX, highest DPPH (50.52 ± 0.03 mg AAE/g extract) and ABTS (26.78 ± 0.03 mg TE/g extract) radical scavenging reported in methanolic extract of fruit; however, acetone extract of leaf showed highest FRAP (376.89 ± 1.95 mg Fe(II)/g extract) and PMA (326.54 ± 4.73 mg AAE/g extract). In contrast, aqueous extract of leaf and fruit revealed highest metal chelating activity (41.67 ± 0.49 mg EDTA/g extract). In anti-diabetic studies, acetonitrile extract of leaves and fruits exhibited appreciable inhibition of α-amylase (83.33%) and α-glycosidase (77.42%) enzymes. Similarly, acetyl cholinesterase (AChE) inhibition was highest in water (88.91%) and acetone (81.87%) extracts of leaf and fruits. Fruit extracts showed potent anticancer activity against breast (MCF-7) and colon (HT-29) cancer cell lines (LC₅₀ 329.36 and 385.17 µg/mL, respectively). RP-HPLC analysis revealed highest cucurbitacin B (CuB) (196.24 ± 1.4 mg/g DW), followed by cucurbitacin I (CuI) and cucurbitacin E (CuE) in the fruits (57.14 ± 4.9 and 2.03 ± 0.03 mg/g DW, respectively). RP-HPLC analysis of extracts revealed presence of gallic acid (GA), catechin (CA), vanillic acid (VA), chlorogenic acid (CHLA) and coumaric acid (COA), in which highest GA found in the fruits (1.26 ± 0.07 mg/g DW). Liquid chromatography and mass spectrometry (LC-MS) and gas chromatography-mass spectrometry (GC–MS) analysis revealed presence of bioactive compounds from various groups. Based on the present findings, it was revealed that the fruit and leaf of L. echinata can be used as potent bioresource for natural antioxidants, anti-diabetic, and anticancer drug.     

Total phenolics,flavonoids and antioxidant properties of three Ceropegia species from Western Ghats of India

The aim of present work was to assess the total phenolic content (TPC), total flavonoid content (TFC), phenolic compounds and antioxidant properties of various extracts of three Ceropegia spp.: Ceropegia spiralis, Ceropegia panchganiensis and Ceropegia evansii from Western Ghats of India. TPC of the samples varied from 0.3 ± 0.2 to 28.5 ± 0.3 mg TAE/g FW, whereas, TFC of the samples ranged between 0.1 ± 0.1 and 15.3 ± 0.3 mg RE/g FW. The major phenolic compounds identified were gallic acid, vanillin, cathechol and ferulic acid. All the extracts possess 1,1-diphenyl-2-picryl hydrazyl (DPPH) radical scavenging activity, ferric reducing antioxidant power (FRAP) as well as metal chelating ability and this was also supported by significant correlation with TPC and TFC. To the best of our knowledge, this is the first paper presenting comprehensive data on TPC, TFC, phenolic profile and antioxidant properties of the Ceropegia spp.     

Influence of plant and environment parameters on phytochemical composition and biological properties of Pistacia atlantica Desf.

The aim of the current study is to analyze the phytochemical, antioxidant and antimicrobial activities of 34 extracts prepared from Pistacia atlantica Desf. subsp. atlantica, according to gender, organ type (roots, buds and fruits), geographical location and stage of ripening. Bud extracts exhibited the highest phenolic content (565.74 ± 9.84 mg GAE/g DM), followed by fruit and root extracts. TFC and TTC ranged from 0.38 ± 0.03 to 1.92 ± 0.11 mg CE/g DM and from 0.37 ± 0.03 to 16.54 ± 0.94 mg CE/g DM, respectively. For DPPH and TAC assays, the values varied from 0.038 ± 0.000 to 1.331 ± 0.114 mg/mL and 1.58 ± 0.06 to 43.64 ± 2.58 mg AAE/g DM, respectively. Besides, bud extracts showed the highest bioactivity against pathogenic bacteria and a slight antifungal effect. Additionally, HPLC-DAD analysis revealed that the caffeic acid and the dimethyl-allyl caffeic acid characterized the bud extract, while the rutin and the hydroxytyrosol were abundant in the red fruit extract. The present evidence suggests that P. atlantica may be considered as a potential source of new additives for therapeutic, food and cosmetic products.     

Preliminary screening of antioxidant and cytotoxic potential of green seaweed,Halimeda opuntia (Linnaeus) Lamouroux

Marine natural products have displayed numerous advantageous effects on biological activities, including antioxidants and cytotoxicity. The total lipids, carotenoids, chlorophyll a and b content, total phenolic content (TPC), total flavonoid content (TFC), and antioxidant activity of methanolic crude extract of the green seaweed Halimeda opuntia were all measured in this study. The TPC of the extracts was determined according to the Folin-Ciocalteu method, yielding a result of 55.04 ± 0.98 mg GAE/g of extract. As determined by the aluminium chloride colorimetric method, the TFC of the extract was 40.02 ± 0.02 mg QE/g of extract. Antioxidant activity was determined by using a 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay with different concentrations that ranged between 200 and 1000 µg/mL, noted H. opuntia as the highest in DPPH reduction (63.61 %) at 1000 µg/mL concentration. Total antioxidant capacity (TAC) of the extract was 57.36 ± 0.004 mg AAE/g extract at concentration of 1.0 mg/mL. The cytotoxic activity of this seaweed was pre-screened against a panel of cell lines including estrogen receptor-positive human breast adenocarcinoma (MCF-7), estrogen negative human breast adenocarcinoma (MDA-MB-231), human colorectal adenocarcinoma (HT-29), human hepatocellular carcinoma (HepG2), and mouse embryonic fibroblast (3T3) using the MTT assay. The content of total lipids in H. opuntia was 1.60 ± 0.002 %. Total carotenoids were 115.57 ± 0.98 µg/g, while chlorophyll a and b were 148.73 ± 2.60 µg/g and 290.83 ± 9.46 µg/g, respectively. In terms of cytotoxicity activity, methanolic extract of H. opuntia was found to be highly cytotoxic to MCF-7 cells, with an IC₅₀ of 25.14 ± 1.02 g/mL, and slightly less so to 3T3 cells (IC₅₀ 65.23 ± 0.25 µg/mL). This study's findings suggest that natural pigments (carotenoids and chlorophyll), phytochemicals like phenolic and flavonoid compounds found in this species may play an important role and could be used as a natural cancer treatment.     

Comparative analysis of antioxidant activity,toxicity, and mineral composition of kernel and pomace of apricot (Prunus armeniaca L.) grown in Balochistan,Pakistan

The present study aims to investigate some physical attributes, total phenolics content, total flavonoids content, mineral composition, bioluminescence toxicity assay and antioxidant activity in terms of DPPH, HPS, TAC and FRAP assays in the kernel and pomace samples of six apricot cultivars grown in Balochistan, Pakistan. TFC and TPC determined by the AlCl₃ and Folin-Ciocalteu assays in apricot kernel extracts of six cultivars varied from 1797.5 (Chagali) to 4778.9 (Badoghur) mg QUE/100 g DW and from 1750.0 (Chagali) to 5005.8 (Badoghur) mg GAE/100 g DW. Apricot kernels exhibited higher antioxidant activity than pomace; antioxidant activity in terms of IC₅₀ in kernels ranged from 24.88 to 98.61 μg/ml for DPPH, 334.84 to 516.63 μg/ml for HPS, from 22.02 to 110.80 μg/ml for TAC and from 96.27 to 163.35 μg/ml for FRAP. The apricot kernels showed higher TPC, TFC, bioluminescence toxicity to V. logei and antioxidant activity than the pomace. The correlation analysis demonstrated substantial contributions of polyphenols and flavonoids to antioxidant assays. The sample type was the leading factor affecting the amounts of K, Na, Ca, Fe, and Mn in the tested samples; mineral contents were higher in pomace than kernels. The highest inhibition to V. logei was found in the kernels of Badoghur (IC₅₀ = 1.61 mg/ml). The PCA analysis showed significant contributions of phenolic and flavonoid contents towards antioxidant bioluminescence toxicity assays. Our results suggest Badoghur, Shakarpara and Sardai kernels are rich sources of secondary metabolites and possess remarkable antioxidant and antiluminescence activity and can make a significant contribution to the treatment and prevention of chronic health problems.     

Phytochemical profiling,antioxidant and antibacterial efficacy of a native Himalayan Fern: Woodwardia unigemmata (Makino) Nakai

Present work elucidates the antioxidant and antibacterial activity of Woodwardia unigemmata (Makino) Nakai along with chemical characterization using its aqueous (AEW), methanol (MEW), and hexane (HEW) extracts. Chemical profile of different extracts was illustrated by using Gas chromatography and mass spectrometry (GC-MS) analysis. Antioxidant activities were tested using DPPH and FRAP assays, total phenolic and flavonoid content by Folin-Ciocalteu and aluminum chloride method, respectively. Further, antibacterial activity against six plant and four animal pathogenic bacteria was analyzed by employing the disc diffusion assay. GC-MS analysis revealed the presence of catechol (21.96%), glycerol (20.22%), n-pentadecanoic acid (6.95%), glyceryl monoacetate (6.35 %), ethyl acetimidate (5.39 %) and 3-hydroxy-2,3-dihydromaltol (5.36%) in AEW; β-sitosterol (17.39%), pentadecanoic acid (9.81%), vitamin E (7.82%) and glycerol (7.05%) in MEW; γ-sitosterol (33.45%), vitamin E (10.04%) and campesterol (7.32%) in HEW as major constituents. Maximum phenolics (873 ± 6.01 mgGAE/g dry extract) as well as flavonoids (151 ± 11.44 mgQE/g dry extract) content was found in MEW, which also showed remarkable antioxidant potential (IC₅₀ 6.07 ± 1.4 µg/ml for DPPH and 768 ± 10.4 mg AAE/g dry extract for FRAP assay. In antibacterial activity, maximum inhibition (15 ± 0.9 mm) was observed for HEW against R. solanacearum, followed by AEW against A. tumefaciens and X. phaseoli (11 ± 0.3 mm each). MEW was found positive only against A. tumefaciens. Significant minimum inhibitory concentration (MIC) value observed for AEW against L. monocytogenes (10 mg/ml). Polar extracts had remarkable antioxidant potential, while non-polar extract did show significant antibacterial activity. Further, GC- MS reports indicated that this traditionally useful fern species can be an excellent source of biologically active compounds.     

Total phenolics, flavonoids and antioxidant activity of Saptarangi (Salacia chinensis L.) fruit pulp

Salacia chinensis L. has various beneficial properties including antidiabetic and antioxidant activity. The S. chinensis fruit pulp (SCFP) was extracted with four different solvents (Methanol, ethanol, acetone and water) and was screened for total phenolic content (TPC), total flavonoid content (TFC) and antioxidant activity (AOA). The AOA was assessed by evaluating the 1, 1-diphenyl-2-picrylhydrazyl (DPPH), ferric reducing antioxidant power (FRAP) and metal chelating assay. Methanolic SCFP extract exhibited the highest phenolic content (3.20?±?0.12 mg GAE/g FW) whereas, ethanolic extract showed highest flavonoid content (0.31?±?0.68 mg RE/g FW). The methanolic extract possesses highest antioxidant activity towards DPPH (92.44 %), FRAP (1.939 O.D) and metal chelating activity (74.16 %). AOA (DPPH and FRAP) was significantly correlated with TPC. The results indicated that SCFP is a good natural source of antioxidant compounds for use in food and pharmaceutical industry.     

Analyses of Elaeocarpus sphaericus Extract for Antioxidant,Antiproliferative and Gene Repression Activities against HIF-1α and VEGF

The study presents antioxidant, phytochemical, anti-proliferative, and gene repression activities against Hypoxia-inducible factor (HIF-1) alpha and Vascular endothelial growth factor (VEGF) of Elaeocarpus sphaericus extract. Elaeocarpus sphaericus dried and crushed plant leaves were extracted using water and methanol by ASE (Accelerated Solvent Extraction) method. Total phenolic content (TPC) and total flavonoid content (TFC) were used to measure the extracts′ phytochemical activity (TFC). Antioxidant potential of the extracts was measured through DPPH, ABTS, FRAP, and TRP. Methanolic extract of the leaves of E. sphaericus has shown a higher amount of TPC (94.666±4.040 mg/gm GAE) and TFC value (172.33±3.21 mg/gm RE). The antioxidant properties of extracts in the yeast model (Drug Rescue assay) showed promising results. Ascorbic acid, gallic acid, hesperidin, and quercetin were found in the aqueous and methanolic extracts of E. sphaericus at varying amounts, according to a densiometric chromatogram generated by HPTLC analysis. Methanolic extract of E. sphaericus (10 mg/ml) has shown good antimicrobial potential against all bacterial strains used in the study except E. coli. The anticancer activity of the extract in HeLa cell lines ranged from 77.94±1.03 % to 66.85±1.95 %, while it ranged from 52.83±2.57 % to 5.44 % in Vero cell lines at varying concentration (1000 μg/ml–31.2 μg/ml). A promising effect of extract was observed on the expression activity of HIF-1 and VEGF gene through RT-PCR assay.     

Profiling the phyto-constituents of Punica granatum fruits peel extract and accessing its in-vitro antioxidant,anti-diabetic,anti-obesity,and angiotensin-converting enzyme inhibitory properties

This context was investigated to assess the in vitro antioxidant, anti-diabetic, anti-obesity, and angiotensin-converting enzyme (ACE) inhibition traits of Punica granatum fruits peel extract. Initially, among various extracts tested, aqueous and ethanolic peel extracts depicted the presence of diverse phytoconstituents. In vitro antioxidative properties of peel extracts were determined using standard methodologies. Results showed that aqueous and ethanolic extracts had IC₅₀ values of 471.7 and 509.16 μg/mL, respectively in terms of 1,1,diphenyl 2,2,picrylhydrazyl scavenging. Likewise, IC₅₀ values of aqueous and ethanol extract were obtained as 488.76 and 478.47 μg/mL towards the degradation of hydrogen peroxide. The ethanolic extract exhibited the highest inhibition of α-glucosidase by showing activity of 53.34 ± 2.0 to 15.18 ± 1.4 U/L in a dose dependent manner (100–1000 µg/mL). Ethanolic extract was reported as the most active inhibitor of lipase with an IC₅₀ value of 603.50 µg/mL. Ethanolic extract showed increased inhibition of ACE in a concentration dependent manner (100–1000 µg/mL) with IC₅₀ value of 519.45 µg/mL. Fourier transform-infrared spectrum revealed the availability of various functional groups in the ethanolic extract of peel. Gas chromatography-mass spectrometry chromatogram of peel extract illustrated 23 diversified chemical constituents including 1,2,3,4-butanetetrol, Dimethyl sulfone, 9-octadecenamide, and Pentadecanoic acid as predominant compounds. In summary, P. granatum fruits peel extract revealed promising antioxidant, anti-diabetic, anti-obesity, and anti-hypertensive properties.     

Phytochemical analysis and bioactivity assessment of five medicinal plants from Pakistan: Exploring polyphenol contents,antioxidant potential,and antibacterial activities

Plants have always been the prime focus in medicine industries due to their enormous ethnobotanical uses and multitude of biological and therapeutic properties. In the current study, preliminary phytochemical composition, Total phenolic content (TPC), and total flavonoid content (TFC) with the antioxidant and antibacterial activity of hydroalcoholic extract and n-hexane, chloroform and n-butanol fractions of five selected medicinal plants [Tephrosia purpurea (L.) Pers., Lavandula stoechas L., Aesculus indica (Wall. ex Cambess.) Hook, Iris ensata Thunb., and Kalanchoe pinnata (Lam.) Pers.] from Pakistan, have been evaluated. TPC and TFC were determined by Folin-Ciocalteu’s and AlCl₃ methods respectively. The antioxidant activity was performed by DPPH, ABTS, FRAP, and CUPRAC while the antibacterial potential of these plants was determined by agar well diffusion assay. K. pinnata (Lam.) Pers. exhibited the highest TPC (695 ± 13.2 mg.GA.Eq.g^-1DE ± SD) in n-butanol fraction and the highest TFC in its chloroform faction (615 ± 6.31 mg Q.Eq.g⁻¹ DE ± SD). The n-butanol fraction and hydroalcoholic extract of I. ensata Thunb. exhibited strong antioxidant potential by DPPH and CUPRAC assays respectively, whereas K. pinnata (Lam.) Pers. n-butanol fraction exhibited the strongest reducing potential. The hydroalcoholic extract of all tested plants exhibited significant antibacterial activity against tested bacterial strains with ZI (12–18 mm). Conclusively, K. pinnata (Lam.) Pers. (Family: Crassulaceae) and I. ensataThunb. (Family: Iridaceae) exhibited the highest antioxidant and antibacterial potential. They can be explored for the isolation of phytoconstituents responsible for this potential and serve as a lead for the production of new natural antioxidants and antibacterial agents that can be used to cure various diseases.     

Antioxidant properties and phenolic profiling by UPLC-QTOF-MS of Ajwah,Safawy and Sukkari cultivars of date palm

Date palm (P. dactylifera) plays a vital role in ethnomedicinal practices in several parts of the world. There are over 2000 cultivars of date palm that differ in chemical composition and extent of bioactivity. The present study was undertaken to comparatively evaluate the antioxidant potential of three cultivars of date palm (Ajwah, Safawy and Sukkari) from Saudi Arabia and analyze their phenolic constituents in order to draw a rationale for their activity. Antioxidant activities of the date cultivars were evaluated by different quantitative methods including 2,2-diphenyl-1-picrylhydrazyl (DPPH) and hydroxyl radical scavenging assay, total antioxidant capacity, reducing power, total phenolic (TPC), flavonoid (TFC) and tannin content (TTC), while qualitative phenolic composition was determined using ultra performance liquid chromatography coupled to quadropole time of flight mass spectrometry (UPLC-QTOF-MS). All the three date extracts showed good DPPH radical scavenging (IC₅₀ 103–177 μg/mL) and hydroxyl radical scavenging (IC₅₀ 1.1–1.55 mg/mL) activity and total antioxidant capacity (IC₅₀ 87–192 μg/mL). The reducing power was also comparable to that of ascorbic acid, used as standard in above experiments. All the three samples contain significant amount of major antioxidant components (phenolic, flavonoid and tannin) that successfully correlates with the results of radical scavenging assays. UPLC-QTOF-MS revealed a total of 22 compounds in these date cultivars classified into common phenolics, flavonoids, sterols and phytoestrogens. Significant variation in the degree of antioxidant activity of these three date cultivars can be attributed to the difference in the content and composition of phenolic compounds.     

Q-TOF LC-MS compounds evaluation of propolis extract derived from Malaysian stingless bees,Tetrigona apicalis,and their bioactivities in breast cancer cell,MCF7

Propolis is known to exhibit various phytochemical compounds that aid in several biological activities. The current study investigates the phytochemical compounds of ethanolic extract of propolis of Tetrigona apicalis (EEP) using Q-TOF LC-MS, its antioxidant properties using DPPH and ABTS⁺ radical scavenging assays, total phenolic (TPC) and flavonoid content (TFC), using Folin-Ciocalteu and Aluminium Chloride method, respectively, as well as proapoptotic effects, based on the selected IC₅₀ of the cytotoxic study conducted for EEP using annexin V-FITC assay. Terpene and polyphenol were among of 17 identified compounds. The EC₅₀ of EEP for DPPH and ABTS⁺ was 1.78 mg/mL and 1.68 mg/mL, while the EEP exhibited TPC and TFC values of 31.99 mgGAE/g and 66.4 mgQCE/g, respectively in which the parameters were strongly correlated. The IC₅₀ of EEP effectively induces apoptosis in MCF7 cells. In conclusion, EEP possessed important phytochemical compounds that work excellently as antioxidants and anticancer agents.     

Analysis of bioactive composites and antiviral activity of Iresine herbstii extracts against Newcastle disease virus in ovo

The study was implemented to actuate the qualitative and quantitative phyto constituents of Iresine herbstii extracts and its antiviral efficacy against avian ND virus. Among four tested solvents, the ethanolic extract of Iresine herbstii revealed the presence of highest quantity of all tested phytochemicals while petroleum ether extract showed the least. Folin-Ciocalteu method assessed the range of TPC extended from 81.01 ± 0.67 to 126.35 ± 0.45 µg GAE/mg. Acetonic extract showed the highest amount among all extracts and petroleum ether possessed the lower quantity. TFC ranged from 54.37 ± 0.45 to 88.12 ± 0.26 µg QE/mg followed by colorimetric method. From all extract ethanolic extract showed highest quantity and petroleum ether revealed the lower. HPLC analysis of ethanolic extract of I. herbstii confesses the presence six bioactive components by using the HP5-MS column. To check the antiviral potential of plants, different prepared treatments of plant extract and live virus were inoculated at 9 days old SPF embryonated chicken eggs. Results exposed that all plant extracts produce antiviral activity against NDV in ovo according to their potential and phytochemical profile. The highest survival rate was observed in the ethanolic extract at 400 µg/mL and acetonic extract at 300 µg/mL as it controls the NDV activity completely, evidence of absence of embryo death and HA titre. Dichloromethane and petroleum ether could not inhibit the virus completely. 600 µg/mL concentration was proved as toxic in all extracts except petroleum ether extract which showed a dose dependent pattern.     

Antioxidant and Proteinase Inhibitory Activities of Selected Poppy (Papaver somniferum L.) Genotypes

Poppy seeds (Papaver somniferum L.) belong to tasty food ingredients however, they should be considered also as valuable source of biologically active compounds. Content of selected metabolites, antioxidant and proteinase inhibitory activities were analyzed in vitro in extracts from seeds of fifteen poppy genotypes. Considerable variation in all parameters was detected within the set of analyzed poppy genotypes. The genotype Major expressed the highest antioxidant activity determined by all four methodological approaches (DPPH, ABTS, FRAP, RP). The genotype MS 423 exhibited the highest inhibitory activities against trypsin, thrombin and collagenase. Very specific position among all had the genotype Redy . Its grain extract reached significantly high levels in 9 out of 14 measured parameters (TPC, TFC, TTC, TAC, FRAP, RP, inhibitory activities against trypsin, thrombin, collagenase). Edible grains of poppy are valuable source of natural compounds which may be beneficial in pathological states associated with oxidative stress or increased proteinase activities.     

Antiangiogenic,anti-inflammatory and their antioxidant activities of Turnera subulata Sm. (Turneraceae)

Turnera subulata is a substantial medicinal plant used in folk medicine to treat various ailments. The current study was assess the total phenolic and flavonoid contents to evaluate the antioxidant and anti-inflammatory activities of the sequentially extracted T. subulata plant samples. In vitro anti-angiogenic activity was evaluated by chick chorioallantoic membrane (CAM) model for chloroform, ethyl acetate and ethanol extracts. The results obtained revealed that total phenolic content of the chloroform extract (24.13 ± 0.27 mg/g) and total flavonoid content (TFC) of the chloroform extract (22.28 ± 0.40 mg/g) were found to be suggestively higher than the other extracts. A strong antioxidant property was observed for all the six extracts. A study anti-inflammatory activity was observed in chloroform and ethanol extracts, with IC₅₀ ranging from 79 ± 1.01 μg/mL to 81 ± 1.01 μg/mL for protein denaturation assay and from 74 ± 0.11 μg/mL to 76 ± 1.11 μg/mL for HRBC membrane stabilization assay, respectively. The chloroform and ethanol extracts have exhibited good antiangiogenic property. Eventually, these results justified that the chloroform and ethanol extracts of T. subulata with great antioxidant, anti-inflammatory and antiangiogenesis potentials could be promising candidates for the development of a cost effective, potent anticancer drug with minimal side effects.     

HPTLC-profiling of eleutherosides,mechanism of antioxidative action of eleutheroside E1, the PAMPA test with LC/MS detection and the structure–activity relationship

Human body is constantly generating free radicals, which causes oxidative stress. Despite naturally occurring antioxidant systems in human body, free radicals cause lipid, proteins and DNA oxidation. New antioxidants are still urgent as well as their mechanisms of action should be explained. In this study, we investigated the mechanism by which eleutherosides B, E and E1 may act as antioxidants, identified eleutherosides in Eleutherococcus lasiogyne and Eleutherococcus giraldii, and explained in vitro the absorption of eleutheroside E1 based on passive transport. The DPPH¹ and DB-HPTLC tests were used to assess the antioxidant activity. Of the three eleutherosides, only eleutheroside E1 exhibited a strong anti-DPPH¹ activity (EC₅₀ 37.03 μg/mL; 63 mMol) compared to the raw extracts (EC₅₀ 170 and 180 μg/mL for E. lasiogyne and E. giraldii). This activity was also confirmed by the DB-HPTLC autography technique. According to Za?uski’s hypothesis, the antioxidant mechanism of eleutheroside E1 is based on the complexation of DPPH¹ molecule with its aryl radical. During this reaction, the aryl radical of eleutheroside E1 (E1¹) and DPPHH are created. Next, the aryl radical (E1¹) is complexed with another DPPH¹ molecule. Additionally, the aryl radical can be stabilized by the presence of the methoxy groups in the aromatic ring, which increases its antioxidative action. The HPTLC-identification of extracts showed the presence of eleutherosides B, E and E1 in both species. The PAMPA test coupled with LC/MS detection showed a low permeability of eleutheroside E1 across artificial membrane. Because eleutherosides belong to the polyphenols, the TPC and TFC were quantified. The TPC and TFC varied from 51.4 to 49.3 mg/g dry extract for TPC, and from 5.73 to 4.91 mg/g dry extract for TFC, for E. giraldii and E. lasiogyne, respectively. In conclusion, eleutheroside E1 in its pure form could be a chemopreventive ingredient of new pharmacological or dietary products, stimulating the GALT. These findings can explain partially the adaptogenic activity of eleutheroside E1 on the GALT, which has been still unknown.     

Free Radical Scavenging Activity and Comparative Metabolic Profiling of In Vitro Cultured and Field Grown Withania somnifera Roots

Free radical scavenging activity (FRSA), total phenolic content (TPC), and total flavonoid content (TFC) of in vitro cultured and field grown Withania somnifera (Ashwagandha) roots were investigated. Withanolides analysis and comprehensive metabolic profiling between 100% methanol extracts of in vitro and field grown root tissues was performed using high performance thin layer chromatography (HPTLC) and gas chromatography-mass spectrometry (GC-MS), respectively. Significantly higher levels of FRSA, TPC, and TFC were observed in in-vitro cultured roots compared with field grown samples. In addition, 30 day-cultured in vitro root samples (1MIR) exhibited a significantly higher FRSA (IC₅₀ 81.01 μg/mL), TPC (118.91 mg GAE/g), and TFC (32.68 mg CE/g) compared with those in 45 day-cultured samples (1.5MIR). Total of 29 metabolites were identified in in vitro cultured and field grown roots by GC-MS analysis. The metabolites included alcohols, organic acids, purine, pyrimidine, sugars, and putrescine. Vanillic acid was only observed in the in vitro cultured root samples, and higher level of the vanillic acid was observed in 1MIR when compared to 1.5MIR. Therefore, it is suggested that 1MIR might serve as an alternative to field grown roots for the development of medicinal and functional food products.     

Antioxidant and Pulmonary Protective Potential of Fraxinus xanthoxyloides Bark Extract against CCl4-Induced Toxicity in Rats

Fraxinus xanthoxyloides is a perennial shrub belonging to family Oleaceae, traditionally used for malaria, jaundice, pneumonia, inflammation, and rheumatism. Our study is aimed to assess the total phenolics (TPC), flavonoids (TFC), terpenoids contents (TTC) and antioxidant profiling of F. xanthoxyloides methanol bark extract (FXBM) and its fractions, hexane, chloroform, ethyl acetate and aqueous, along with high-performance liquid chromatography with diode-array detection (HPLC-DAD). Further, the antioxidant and pulmonary protective potential was explored against carbon tetrachloride (CCl₄)-induced CCl4-induced pulmonary tissue damage in rats. The highest TPC, TFC and TTC were found in FXBM (133.29±4.19 mg/g), ethyl acetate fraction (279.55±10.35 mg/g), and chloroform fraction (0.79±0.06 mg/g), respectively. The most potent antioxidant capacity was depicted by FXBM (29.21±2.40 μg/mg) and ethyl acetate fraction (91.16±5.51 μg/mg). The HPLC-DAD analysis revealed the predominance of gallic, chlorogenic, vanillic and ferulic acid in FXBM. The administration of CCl₄ induced oxidative stress, suppressed antioxidant enzymes′ activities including catalase, peroxidase, superoxide dismutase, glutathione peroxidase, glutathione-S-transferase, and glutathione reductase. Further, it increased thiobarbituric acid reactive substances (TBARS) and H₂O₂ levels, induced DNA injuries and reduced the total protein and glutathione content in lung tissues. The treatment of rats with FXBM restored these biochemical parameters to the normal level. Moreover, the histopathological studies of lung tissues demonstrated that FXBM protected rats′ lung tissues from oxidative damage restoring normal lung functions. Thus, F. xanthoxyloides bark extract is recommended as adjuvant therapy as protective agent for patients with lung disorders.     

Phytochemical analysis,antioxidant and antimicrobial activity of wild and in vitro derived plants of Ceropegia thwaitesii Hook – An endemic species from Western Ghats,India

Ceropegia thwaitesii Hook (Asclepiadaceae), an endemic plant species, due to habitat destruction and over exploitation has a very restricted distribution in the Western Ghats of Tamil Nadu, India. The present wrok aimed to determine the chemical composition, the total phenolic (TPC), flavonoid (TFC) and tannin content (TEC), and to assess the antioxidant properties of various extracts of in vivo plants (IVP) and in vitro regenerated plants (IRP) of C. thwaitesii. Some phenolic compounds like gallic acid, cathechol, vanillin and salicylic acid were identified and quantified by HPLC. All the extracts possessed relevant radical scavenging activity on DPPH, Superoxide radical scavenging activity, and Nitric oxide radicals as well as total antioxidant ability. DPPH assay of in vitro methanol stems extracts and ethanol leaves extracts revealed the best antioxidant properties with important IC₅₀ values of 0.248?±?0.45?µg/mL and 0.397?±?0.67?µg/mL, respectively, whereas in vivo chloroform stems extracts showed a lower antioxidant activity (IC₅₀ of 10.99?±?0.24?µg/mL). The IRP methanol extracts of stem and leaves had good inhibitory activity against all tested microorganisms in a dose-dependent manner. These results suggested that in vitro raised plants of C. thwaitesii are an excellent source of antioxidant compounds to be exploited on an industrial level as food additive.     

A new isoflavonol and other constituents from Cameroonian propolis and evaluation of their anti-inflammatory,antifungal and antioxidant potential

Propolis is rich in diverse bioactive compounds. Propolis samples were collected from three localities of Cameroon and used in the study. Column chromatography separation of propolis MeOH:DCM (50:50) extracts yielded a new isoflavonol, 2-hydroxy-8-prenylbiochanin A (1) alongside 2′,3′-dihydroxypropyltetraeicosanoate (2) and triacontyl p-coumarate (3) isolated from propolis for first time together with seven compounds: β-amyrine (4), oleanolic acid (5), β-amyrine acetate (6), lupeol (7), betulinic acid (8), lupeol acetate (9) and lupenone (10). These compounds were tested for their inhibitory effect on oxidative burst where intracellular reactive oxygen species (ROS) were produced from zymosan stimulated human whole blood phagocytes and on production of nitric oxide (NO) from lipopolysaccharide (LPS) stimulated J774.2 mouse macrophages. The cytotoxicity of these compounds was evaluated on NIH-3 T3 normal mouse fibroblast cells, antiradical potential on 2,2-diphenyl-1-picrylhydrazylhydrazyl (DPPH·) as well as their anti-yeast potential on four selected candida species. Compound 1 showed higher NO inhibition (IC₅₀ = 23.3 ± 0.3 µg/mL) than standard compound L-NMMA (IC₅₀ = 24.2 ± 0.8 µg/mL). Higher ROS inhibition was shown by compounds 6 (IC₅₀ = 4.3 ± 0.3 µg/mL) and 9 (IC₅₀ = 1.1 ± 0.1 µg/mL) than Ibuprofen (IC₅₀ = 11.2 ± 1.9 µg/mL). Furthermore, compound 1 displayed moderate level of cytotoxicity on NIH-3 T3 cells, with IC₅₀ = 5.8 ± 0.3 µg/mL compared to the cyclohexamide IC₅₀ = 0.13 ± 0.02 µg/mL. Compound 3 showed lower antifungal activity on Candida krusei and Candida glabrata, MIC of 125 μg/mL on each strain compared to 50 μg/mL for fuconazole. The extracts showed low antifungal activities ranging from 250 to 500 μg/mL on C. albicans, C. krusei and C. glabrata and the values of MIC on Candida parapsilosis were 500 μg/mL and above. DPPH* scavenging activity was exhibited by compounds 1 (IC₅₀ = 15.653 ± 0.335 μg/mL) and 3 (IC₅₀ = 89.077 ± 24.875 μg/mL) compared to Vitamin C (IC₅₀ = 3.343 ± 0.271 μg/mL) while extracts showed moderate antiradical activities with IC₅₀ values ranging from 309.31 ± 2.465 to 635.52 ± 11.05 µg/mL. These results indicate that compounds 1, 6 and 9 are potent anti-inflammatory drug candidates while 1 and 3 could be potent antioxidant drugs.