Identification of an isoflavone glycoside hydrolyzing b-glucosidase in the seeds of the desert legume Prosopis cineraria

2017 ◽  
Author(s):  
V. Asati ◽  
P. K. Sharma
Keyword(s):  
High Performance ◽  
Arid Zones ◽  
Prosopis Cineraria ◽  
Methanolic Extracts ◽  
Isoflavone Glycoside ◽  
Microbe Interactions ◽  
Metabolic Screening ◽  
Glycoside Hydrolysis ◽  
Layer Chromatography ◽  
Temperature Optima

Legumes, in general, are rich in glycosidic isoflavone conjugates, whereby specific b-glucosidases act to release the free aglycones that could serve in plant-microbe interactions and defense. Prosopis cineraria is an important legume tree, bearing medicinally useful and edible pods, generally growing in extreme arid/semi-arid zones of Rajasthan. However, specific phytochemical-cum-metabolic profiles are lacking for the same. Therefore, the present investigation undertook phytochemical and metabolic screening of the pods/seeds of P. cineraria for the presence of putative isoflavonoids viz. genistein and daidzein, their glycosides and b-glucosidase(s) capable of catalyzing the glycoside hydrolysis. Extraction and identification of these two aglycone forms of the above isoflavonoids were performed with solvent partition chromatography and Fluorescent/High Performance Thin Layer Chromatography, respectively. Furthermore, optimization of the isoflavone conjugate-specific b-glucosidase activity with respect to pH optima, temperature and time were carried out. The partially purified enzyme showed a temperature optima of 50°C and pH optima of 4.5. The enzyme also demonstrated activity towards natural substrates daidzin and genistin which are glycosides of isoflavonoids daidzein and genistein respectively. The methanolic extracts of the seeds of P. cineraria indicated the presence of related isoflavonoids which needs to be further validated.

scholarly journals Physiologically Active Compounds in Four Species of Phellinus

2017 ◽  
Vol 12 (3) ◽  
pp. 1934578X1701200 ◽  
Author(s):  
Katarzyna Sułkowska-Ziaja ◽  
Anna Maślanka ◽  
Agnieszka Szewczyk ◽  
Bożena Muszyńska
Keyword(s):  
Phenolic Acids ◽  
High Performance ◽  
Total Content ◽  
Dry Weight ◽  
Hplc Method ◽  
Indole Compounds ◽  
Methanolic Extracts ◽  
Free Phenolic Acids ◽  
Layer Chromatography ◽  
Physiologically Active Compounds

The content of two groups of compounds with biological activity (non-hallucinogenic indole compounds and free phenolic acids) were analyzed in extracts of fruiting bodies of four species of Phellinus: P. igniarius, P. pini, P. pomaceus and P. robustus. The presence of indole compounds in methanolic extracts was analyzed by high-performance liquid chromatography and thin-layer chromatography coupled with densitometric detection. Three metabolites (serotonin, tryptamine, and L-tryptophan) were identified. The contents of individual indole compounds ranged from 1.70 (tryptamine in P. robustus) to 8.32 mg x 100 g1 dry weight (L-tryptophan in P. robustus). Four free phenolic acids were detected in methanolic extracts by the HPLC method. The total content ranged from 9.9 mg x 100 g1 DW (P. igniarius) to 32.5 mg x 100 g1 DW (P. robustus).

scholarly journals HPTLC METHOD FOR ESTIMATION AND QUANTIFICATION OF β-SITOSTEROL FROM IN-VIVO AND IN-VITRO SAMPLES OF MERREMIA AEGYPTIA AND MERREMIA DISSECTA

2020 ◽  
pp. 162-165
Author(s):  
RIDHI JOSHI ◽  
RISHIKESH MEENA ◽  
PREETI MISHRA ◽  
VIDYA PATNI
Keyword(s):  
High Performance ◽  
Normal Phase ◽  
Leaf Sample ◽  
Plant Parts ◽  
Methanolic Extracts ◽  
Aluminum Plates ◽  
Layer Chromatography ◽  
Hptlc Method

Objective: A normal-phase high-performance thin-layer chromatography (HPTLC) method has been developed and validated for estimation and quantitation of beta-sitosterol from the methanolic fraction of different plant parts of two medicinally important plants viz. Merremia aegyptia and Merremia dissecta. These plants have been reported to possess antimicrobial, antioxidant, and anti-inflammatory activities. Methods: Chromatographic separation of beta-sitosterol from the methanolic extracts of plant parts of M. aegyptia and M. dissecta was performed on TLC aluminum plates pre-coated with silica gel 60F254 using a suitable mobile phase. The densitometric scanning was done after derivatization at ????-580 nm for ????-sitosterol. Result: Only M. dissecta leaf sample was reported to contain ????-sitosterol (4.6 ng/μl), whereas other samples such as seed, stem, and callus extracts of M. aegyptia and M. dissecta did not showed its presence. Conclusion: The developed HPTLC method is simple, rapid, and precise and can be used for routine analysis and quantification of ????-sitosterol and other useful plant bioactives that are phytopharmaceutically important.

Determination of α-, β and γ-Amanitin by High Performance Thin-Layer Chromatography in Amanita phalloides (Vaill. ex Fr.) Seer, from Various Origin

1979 ◽  
Vol 34 (12) ◽  
pp. 1133-1138 ◽  
Author(s):  
Tjakko Stijve ◽  
Ruth Seeger
Keyword(s):  
Thin Layer ◽  
Thin Layer Chromatography ◽  
High Performance ◽  
Limit Of Detection ◽  
Dry Weight ◽  
Amanita Phalloides ◽  
Prolonged Storage ◽  
Methanolic Extracts ◽  
Layer Chromatography

A fast, sensitive high performance thin-layer chromatographic method for the determination of α-, β-, and γ-amanitin in crude, methanolic extracts of Amanita phalloides is described. The limit of detection is 50 ng of each amanitin. With this method amanitin was determined in 24 pooled samples of Amanita phalloides, collect­ed between 1970 and 1977 in Germany and Switzerland. The total amanitin content varied be­tween 2010 and 7300 mg/kg dry weight and the average value was 4430 mg/kg of which 43% was α-amanitin, 49% β-amanitin and 8% γ-amanitin. The origin of the fungi hardly influenced their amanitin content: in samples collected during the same year at different sites it fluctuated within a factor of 1.7. The amanitin content of samples from the same site, but collected in different years, maximally varied within a factor of 3.7. The partial decomposition of amanitins during prolonged storage of the lyophilized samples undoubtedly contributed to this variation. Phalloidin, which was determined by conventional thin-layer-chromatography, could not be de­tected in a sample from 1970, whereas its concentration in material collected during 1977 amount­ed to 2400 mg/kg dry weight. The toxicity of the samples (LD50 of lyophilized defatted methanolic extracts intravenously for mice) varied within a factor of 2.5.

scholarly journals Compounds from Vernonia arborea Buch.-Ham. Inhibit Microbes that Impair Wound Healing

2021 ◽  
pp. 103-113
Author(s):  
Lalitha Vaidyanathan ◽  
T. Sivaswamy Lokeswari
Keyword(s):  
Wound Healing ◽  
High Performance ◽  
Healing Process ◽  
Bioactive Compound ◽  
Wound Management ◽  
Methanolic Extracts ◽  
Agar Well Diffusion Assay ◽  
Microtiter Assay ◽  
Layer Chromatography ◽  
Antimicrobial Potency

Aims: To identify the antimicrobial potency of the leaf fractions of Vernonia arborea against selected wound microbes viz., Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa, Klebsiella pneumoniae and Stenotrophomonas maltophilia. Background: Wound healing is often delayed due to the presence of polymicrobial load, that have to be abolished to facilitate the healing process. A major class of antimicrobial phytocompound reported to occur in Vernonia arborea species include sesquiterpenes. Reports on the wound healing potency of V. arborea in wound models of Wistar rats however did not report antimicrobial activity of the aqueous or methanolic extracts. Methodology: The column fractions of the hexane leaf extract were tested against the selected strains by agar well diffusion assay and the zone of inhibition confirmed with TLC bioautography at specific Rf. The minimum inhibitory concentration (MIC) of the bioactive fractions was identified using resazurin microtiter assay (REMA) and the minimum bactericidal concentration (MBC) was determined. HPTLC quantification was also performed. Results: Out of the 30 pooled fractions, six showed antimicrobial potency against all the five tested wound microbes. The minimum inhibitory concentrations of these fractions were determined, ranging from 15.62 µg/mL to 500 µg/mL for the different microbes. Quantitative High-Performance Thin Layer Chromatography (HPTLC) revealed two compounds (a and b) in the bioactive fraction10 with yields of 633 mg (63%) and 97 mg (9.7%) per gram of the extract. Conclusion: The findings suggest the potential use of the bioactive compound in chronic infectious wound management therapy.

scholarly journals LUPEOL VALIDATION AND QUANTIFICATION IN HETEROPOGON CONTORTUS (L.) BEAUV. (SPEAR GRASS) THROUGH HIGH-PERFORMANCE THIN-LAYER CHROMATOGRAPHY

2017 ◽  
Vol 10 (12) ◽  
pp. 392 ◽  
Author(s):  
Navjot Kaur ◽  
Raghbir Chand Gupta
Keyword(s):  
Thin Layer ◽  
Thin Layer Chromatography ◽  
High Performance ◽  
Marker Compound ◽  
Heteropogon Contortus ◽  
Methanolic Extracts ◽  
Validation And Quantification ◽  
Two Samples ◽  
Layer Chromatography

Objective: This is aimed to study the chromatographic evaluation of triterpenoid, i.e., lupeol from methanolic extracts of leaves, stem, and inflorescence of Heteropogon contortus.Methods: The high-performance thin-layer chromatography (HPTLC) densitometry determination of lupeol was performed using optimized mobile phase toluene:methanol:formic acid (7:3:0.3 v/v) with a derivatization of freshly prepared anisaldehyde-sulfuric acid. For densitometry measurements, the plates were scanned at 530 nm absorbance/reflectance wavelength. Quantification of lupeol marker compound in H. contortus leaves, stem, and inflorescence is estimated using 2-12 μg/spot.Results: The appearance of light purple bands on the chromatograms confirmed the lupeol component in plant samples. Further, the confirmation of the compound is done from the densitometric scanning by comparing λmax values. From this, it is reported that lupeol is present in leaf samples, i.e., 10 mg/g of dry wt., while in rest of the two samples, it is found absent.Conclusion: The leaves of H. contortus (spear grass) are a good source of lupeol and can be used as an alternate natural source to synthesize herbal drugs to control cancer and other anti-inflammatory agents. The presently selected HPTLC is validated and most accurate for the quantification and identification of lupeol present in the selected plant. The leaves of the species which are rich in lupeol can be used in pharmaceutical industry.

scholarly journals Detection and Estimation of alpha-Amyrin, beta-Sitosterol, Lupeol, and n-Triacontane in Two Medicinal Plants by High Performance Thin Layer Chromatography

2014 ◽  
Vol 2014 ◽  
pp. 1-7 ◽  
Author(s):  
Saikat S. Mallick ◽  
Vidya V. Dighe
Keyword(s):  
Thin Layer ◽  
Thin Layer Chromatography ◽  
High Performance ◽  
Simultaneous Estimation ◽  
Pluchea Lanceolata ◽  
Methanolic Extracts ◽  
Whole Plant ◽  
Plant Powders ◽  
Layer Chromatography ◽  
Hptlc Method

A normal phase high performance thin layer chromatography (HPTLC) method has been developed and validated for simultaneous estimation of four components, namely, alpha-amyrin, beta-sitosterol, lupeol, and n-triacontane from two medicinally important plants, Leptadenia reticulata Wight & Arn. and Pluchea lanceolata (DC.) CB. Clarke. In Ayurveda, both plants have been reported to possess immunomodulatory activity. Chromatographic separation of the four components from the methanolic extracts of whole plant powders of Leptadenia reticulata Wight & Arn. and Pluchea lanceolata (DC.) CB. Clarke. was performed on TLC aluminium plates precoated with silica gel 60F254 using a suitable mobile phase. The densitometric scanning was done after derivatization at λ = 580 nm for α-amyrin, β-sitosterol, and lupeol, and at 366 nm for n-triacontane. The developed HPTLC method has been validated and used for simultaneous quantitation of the four components from the methanolic extracts of whole plant powders of Leptadenia reticulata Wight & Arn. and Pluchea lanceolata (DC.) CB. Clarke. The developed HPTLC method is simple, rapid, and precise and can be used for routine quality control.

scholarly journals Quantitative Analysis of Rutin by HPTLC and In Vitro Antioxidant and Antibacterial Activities of Phenolic-Rich Extracts from Verbesina sphaerocephala

Plants ◽  
2021 ◽  
Vol 10 (3) ◽  
pp. 475
Author(s):  
Kathia Yanelly Rodríguez-Valdovinos ◽  
Rafael Salgado-Garciglia ◽  
Monserrat Vázquez-Sánchez ◽  
Dioselina Álvarez-Bernal ◽  
Ernesto Oregel-Zamudio ◽  
...  
Keyword(s):  
Antimicrobial Activity ◽  
Antioxidant Capacity ◽  
High Performance ◽  
Antibacterial Activities ◽  
Wild Plants ◽  
Methanolic Extracts ◽  
Layer Chromatography ◽  
Phenolic And Flavonoid ◽  
And Staphylococcus Aureus

Verbesina sphaerocephala A. Gray, like other wild plants of the genus Verbesina, has been used in herbal medicine. There is information for other species of the genus related to their phenolic content, antioxidant capacity, and isolation of bioactive compounds with antimicrobial activity. However, there are no reports for V. sphaerocephala, although it has an important presence in the state of Michoacán, México. In this study, the phenolic composition, quantification of rutin, and in vitro antioxidant and antibacterial activities of methanolic extracts from V. sphaerocephala leaves and flowers were determined. The results showed that all the investigated extracts have high phenolic and flavonoid contents. The flavonoid rutin was identified in all the extracts from V. sphaerocephala by high-performance thin-layer chromatography (HPTLC). The V. sphaerocephala extracts showed scavenging activity against DPPH• and ABTS•+ radicals (IC50 and 5.83 ± 0.50 and 0.93 ± 0.01 mg/mL, respectively) as well as relevant antioxidant capacity (51.05 ± 0.36 mg of ascorbic acid/g of dry tissue). The experimental results show that V. sphaerocephala extracts possessed a strong antimicrobial activity against Escherichia coli and Staphylococcus aureus bacteria. This research indicates that V. sphaerocephala could be considered as a potential source of natural compounds from the point of ethnopharmacological usage.

scholarly journals HPTLC analysis of berberine in stem extracts of Fibraurea darshani

2018 ◽  
Vol 7 (2) ◽  
pp. 2021
Author(s):  
Sheema Dharmapal ◽  
Bindu T.K. ◽  
Elyas K.K.
Keyword(s):  
High Performance ◽  
Uv Light ◽  
Solvent System ◽  
Plant Sample ◽  
Phytochemical Analysis ◽  
Methanolic Extracts ◽  
Uv Reflectance ◽  
The Family ◽  
Rf Values ◽  
Layer Chromatography

The present study is a first report on the phytochemical analysis of the plant Fibraurea darshani which is endemic to Western Ghats. The plant is a woody dioecious climber belonging to the family Menispermaceae. Preliminary phytochemical screening of methanolic extracts of the stem of F. darshani revealed the presence of secondary metabolites like alkaloids, carbohydrates, anthraquiones, terpenoids, flavonoids, phenolics, sterols etc. A simple and reproducible high performance thin layer chromatography was developed to evaluate the presence of berberine in methanol extract of stem of F. darshani. This method involves separation of compounds by HPTLC on pre-coated silica gel 60F 254 plates with a solvent system of Chloroform: Ethyl acetate: Methanol: Formic acid (4:5:4:0.3) and scanned using densitometric scanner in UV reflectance photo mode at 254 and 366nm. The Rf values (0.97) for berberine in the plant sample and the reference standard were found comparable under UV light at 366nm. The HPTLC method developed was simple, accurate and specific.

scholarly journals HPTLC Fingerprints of Various Secondary Metabolites in the Traditional Medicinal Herb Hypochaeris radicata L.

2016 ◽  
Vol 2016 ◽  
pp. 1-11 ◽  
Author(s):  
Jamuna Senguttuvan ◽  
Paulsamy Subramaniam
Keyword(s):  
Secondary Metabolites ◽  
Western Ghats ◽  
High Performance ◽  
Leaf Extract ◽  
Root Extract ◽  
Root Extracts ◽  
Methanolic Extracts ◽  
Hypochaeris Radicata ◽  
Layer Chromatography ◽  
The Western Ghats

The aim of this work was to elucidate the various secondary metabolites such as alkaloids, flavonoids, glycosides, saponins, and terpenoids in the methanolic leaf and root extracts of Hypochaeris radicata, a most important traditional medicinal plant species in Nilgiris, the Western Ghats, India, using high performance thin layer chromatography (HPTLC). This study was carried out using CAMAG HPTLC system equipped with LINOMAT 5 applicator, TLC scanner 3, Reprostar 3, and winCATS 1.3.4 software. A comprehensive assortment of phytoconstituents in methanolic extracts through HPTLC fingerprinting profiles displayed the existence of alkaloids (3 in leaf and 1 in root extract), flavonoids (4 in leaf extract and 5 in root extract), glycosides (1 in leaf extract and 3 in root extract), saponins (1 in root extract), and terpenoids (1 in leaf and root extracts, resp.). The current study overlays boulevard for H. radicata to provide a direction for further exploration in precluding communicable and noncommunicable ailments.

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