Biochemical study of root extract of Withania somnifera (L.)plant through HPLC analysis

Author(s):  
Naveen Gaurav ◽  
Arun Kumar ◽  
Aditi Grover ◽  
Deepak Som ◽  
U. K. Chauhan ◽  
...  

Secondary metabolite contents of W. somnifera varied remarkably between seasons and genotypes under ex vitro condition. In vitro studies provide an optimum culture condition for steady and quality production of bioactive chemicals throughout the year without involvement of environmental stresses. Mass production of micro-shoots and plantlets by exploring the organogenic totipotency of shoot tip explants (ex vitro and in vitro grown) considering two elite genotypes (Poshita and Jawahar 22) of W. somnifera, and assessment of their capability in production and accumulation of bioactive metabolites (total alkaloid and withanolides amount were quantified; withanolide A and withaferin A contents were estimated by High Performance Liquid Chromatography-HPLC).

2018 ◽  
Vol 4 (3) ◽  
pp. 114-119
Author(s):  
Quang Ung Le ◽  
◽  
Horng Liang Lay ◽  
Ming Chang Wu ◽  
◽  
...  

Agrimonia pilosa Ledeb (AL) has received considerable attention as a herbal medicine for its applications in ethnopharmacology with heath benefits. This study aimed to investigate antioxidant activities and A549 growth inhibitory capacity from its root extract (RE) and aerial parts extract (AE). The 50% ethanol extracts were used for the tests. The total polyphenol content and the antioxidant effects comprising ABTS+ and DPPH free radical scavenging activities were evaluated. Phenolic compounds in the extracts were isolated using high performance liquid chromatography (HPLC). Lactate dehydrogenase released in medium was also evaluated. Total phenolic and flavonoid content, and in vitro antioxidant potential of the RE were higher (p<0.01) than that of the AE. Two individual phenolic compounds consisting of 4-hydroxybenzoic acid and p-coumaric acid were firstly identified in both by HPLC. The RE exhibited higher A549 inhibitory capacity compared to the AE and activated the apoptotic proteins of bcl-2, bax, bad, caspase-3 and caspase-9 in A549. In conclusion, the AL extracts were more effective in antioxidant and A549 cells inhibitory capacity.


2008 ◽  
Vol 63 (5-6) ◽  
pp. 409-412 ◽  
Author(s):  
Farzana Sabir ◽  
Neelam S. Sangwan ◽  
Narayan D. Chaurasiya ◽  
Laxmi N. Misra ◽  
Rajender S. Sangwan

In vitro multiple shoots, root, callus and cell suspension cultures of Withania somnifera exhibited the potentiality to produce pharmacologically active withanolides. Multiple shoots cultures exhibited an increase in withanolide A accumulation compared to shoots of the mother plant. In vitro generated root cultures as well as callus and suspension cultures also produced withanolides albeit at lower levels.


Author(s):  
Somanatha Jena ◽  
Ram C. Jena ◽  
Rasmita Bhol ◽  
Khusbu Agarwal ◽  
Ansuman Sarangi ◽  
...  

<p><strong>Objective: </strong>The present investigation explores the possibilities of using the <em>in vitro</em> and <em>in vivo </em>root and leaf extracts of <em>Withania somnifera </em>for anti-diabetic and anti-hyperlipidaemic effects on streptozotocin-induced diabetic rats.</p><p><strong>Methods: </strong><em>In vitro </em>shoot cultures of <em>Withania somnifera</em> were raised by the axillary proliferation in nodal explants from a garden grown plant using Murashige and Skoog medium then <em>in</em><em> vitro</em> raised roots and shoots were used for the anti-hyperglycemic and anti-hyperlipidaemic experiment. After 72 h of STZ administration, the fasting blood glucose levels were measured and the rats showing FBG level&gt;220 mg/dl were considered to be diabetic and were used for the hyperglycemic study. <em>In vitro</em> and <em>in vivo</em> methanolic root and leaf extracts were orally administered daily to diabetic rats for eight weeks. After the treatment period, blood glucose and serum enzymes like aspartate transaminase (AST), alanine transaminase (ALT), alkaline phosphatase (ALP), total cholesterol, triglycerides, HDL-c high density lipoprotein-bound cholesterol, LDL-c low density lipoprotein-bound cholesterol, LDH, serum protein level, total phenolics and anti-oxidative analysis (DPPH and FRAP) were determined.</p><p><strong>Results: </strong>The levels of blood glucose, AST, ALT, ALP, LDH, HDL-c significantly increased by the use of <em>in vitro</em> methanolic root extracts compared to normal control rats. However, remarkable loss of total protein, albumin, albumin: globulin (A: G) ratio was reported in streptozotocin-induced diabetic rats by using <em>in vitro</em> root extracts. Methanolic <em>in vitro</em> root extract at the dose levels of 300 mg/kg body weight produced a significant decrease in fasting blood glucose (FBG) level by 102.65 with respect to initial fasting blood glucose level after 30 d of the treatment. <em>In vitro</em> root extract demonstrated highest DPPH and FRAP free radical scavenging activity, i.e. 86.55±1.77 and 48.87±2.55 than other extracts.</p><p><strong>Conclusion: </strong>It may be concluded that methanolic <em>in vitro</em> root extract <em>W. somnifera </em>at the dose (300 mg/kg) has more potent anti-hyperglycaemic activity than the other <em>in vitro</em> and <em>in vivo </em>extracts of leaf and root on streptozotocin induced diabetic rats and was also found to be similar in effect to that of the standard drug ‘Glibenclamide’.</p>


2020 ◽  
Vol 26 (4) ◽  
pp. 281-291
Author(s):  
Natália Carolina Vieira ◽  
Patrícia Cardoso Cortelo ◽  
Ian Castro-Gamboa

Fungi are an important source of natural products found in a variety of plant species. A wide range of methods for the detection of metabolites present in fungi have been reported in the literature. The search for methodologies that allow the rapid detection of compounds present in crude extracts is crucial to enable the metabolite annotation doing a qualitative analysis of the complex matrix. Mass spectrometry is an important ally when it comes to in silico detection of previously reported metabolites. In this work, the ethyl acetate extract of Fusarium solani was analyzed by gas chromatography coupled to mass spectrometry (GC/MS) after derivatization process. The ethyl acetate extract was also investigated by liquid chromatography coupled with high-resolution tandem mass spectrometry assisted by the UNIFI software system. A library containing previously reported metabolites from the Fusarium genus was added to the UNIFI platform. Simultaneously, the extract was analyzed through anticholinesterase and antifungal assays. The analysis of the derivatized extract by GC/MS led to the putative identification of five metabolites, and the investigation using Ultra-High Performance Liquid Chromatography - Quadrupole Time-of-Flight Mass Spectrometry (UPLC-QTOF) analysis in data-independent acquisition mode (mass spectrometry) led to the annotation of 15 compounds present in the built-in Fusarium library added to the UNIFI system. The Fusarium solani extract showed potential anticholinesterase and in vitro antifungal activity supported by the detection of bioactive metabolites.


2018 ◽  
Vol 96 (4) ◽  
pp. 433-440 ◽  
Author(s):  
T.R. Anju ◽  
S. Smijin ◽  
M. Jobin ◽  
C.S. Paulose

Temporal lobe epilepsy involves a sequence of events that can lead to neurotransmitter signalling alterations. There are many herbal extracts considered to be alternative therapeutic methods to manage epilepsy. In this study, we investigated the effect of Withania somnifera (WS) root extract and withanolide A (WA) in the management of temporal lobe epilepsy. Confocal imaging of TOPRO-3-stained cortical sections showed severe damage in the epileptic brain. We also observed a reduced antioxidant potential and increased peroxide levels in the epileptic test group of rats. Oxidative stress resulted in the down-regulation of CREB, NF-κB, and TNF-α, and with up-regulation of the apoptotic factors caspases 8 and 3 and Bax in the epileptic group. Epileptic condition also resulted in increased muscarinic receptor binding and mRNA expression in the cerebral cortex. Withania somnifera and withanolide A significantly reversed the altered muscarinic receptor expression and reversed the oxidative stress and resultant derailment in cell signalling. Thus our studies suggest that Withania somnifera and withanolide A play important roles in central muscarinic receptor functional balance and activation of the antioxidant system in the cerebral cortex in temporal lobe epilepsy. These findings can be of immense therapeutic significance for managing epilepsy.


2018 ◽  
Vol 4 (3) ◽  
pp. 120-122
Author(s):  
M.N.F. Rizniya ◽  
◽  
A.H.M. Mawjood ◽  
L.D.A.M. Arawwawala ◽  
◽  
...  

Agrimonia pilosa Ledeb (AL) has received considerable attention as a herbal medicine for its applications in ethnopharmacology with heath benefits. This study aimed to investigate antioxidant activities and A549 growth inhibitory capacity from its root extract (RE) and aerial parts extract (AE). The 50% ethanol extracts were used for the tests. The total polyphenol content and the antioxidant effects comprising ABTS+ and DPPH free radical scavenging activities were evaluated. Phenolic compounds in the extracts were isolated using high performance liquid chromatography (HPLC). Lactate dehydrogenase released in medium was also evaluated. Total phenolic and flavonoid content, and in vitro antioxidant potential of the RE were higher (p<0.01) than that of the AE. Two individual phenolic compounds consisting of 4-hydroxybenzoic acid and p-coumaric acid were firstly identified in both by HPLC. The RE exhibited higher A549 inhibitory capacity compared to the AE and activated the apoptotic proteins of bcl-2, bax, bad, caspase-3 and caspase-9 in A549. In conclusion, the AL extracts were more effective in antioxidant and A549 cells inhibitory capacity.


2007 ◽  
Vol 55 (9) ◽  
pp. 1371-1375 ◽  
Author(s):  
Rajender Singh Sangwan ◽  
Narayan Das Chaurasiya ◽  
Payare Lal ◽  
Laxminarain Misra ◽  
Girish Chandra Uniyal ◽  
...  

2020 ◽  
Vol 15 ◽  
pp. 263310552093769 ◽  
Author(s):  
Priya Kashyap ◽  
Heera Ram ◽  
Sunil Dutt Shukla ◽  
Suresh Kumar

Alzheimer’s disease (AD) is characterized by depositions of amyloid β (Aβ) peptides aggregates resulting in plaques formation in the central nervous system (CNS). This study evaluates the disease-modifying potential of scopoletin against multiple factors associated with AD such as cholinesterase enzymes, Aβ peptides, and neuroprotective properties against Aβ- and H2O2-induced cytotoxicity under in vitro conditions. Scopoletin was identified and quantified using UPLC-QTOF (ultra-high performance liquid chromatography-quadrupole time-of-flight) and high-performance liquid chromatography (HPLC), respectively. The antiamyloidogenic potential was evaluated by thioflavin T and congo red binding assay. Inhibition of key enzymes, that is, acetylcholinesterase and butyrylcholinesterase, was investigated by Ellman’s assay. UPLC-QTOF analysis showed that most abundant phytoconstituent present in Argyreia speciosa hydroalcoholic root extract was scopoletin followed by festuclavine and ergometrine. Scopoletin was further quantified using novel reverse phase (RP)-HPLC method developed in this study. The neuroprotective potential of scopoletin was found to be 69% against Aβ42-induced neurotoxicity and 73% against H2O2-induced cytotoxicity in PC12 cell culture at 40 μM final concentration. At the same concentration, scopoletin inhibited Aβ42 fibril formation up to 57%. The IC50 concentration for AChE and BuChE enzyme inhibition by scopoletin was 5.34 and 9.11 μM, respectively. The antiaggregation and enzyme inhibition results were complemented with strong molecular interactions of scopoletin with target proteins validated by in silico molecular docking analysis. Based on this study, it can be concluded that scopoletin can be used as a lead for amelioration of symptoms and disease-modifying effects in AD.


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