LC-MS/MS method for the quantification of withaferin-A in plant extracts of<i>Withania</i>spp.

Mirjalili, Mohammad Hossein; Navarro, Anna; Hernández, Luís E.; Jáuregui, Olga; Bonfill, Mercedes

doi:10.1556/achrom.25.2013.4.12

Cited by 7 publications

(6 citation statements)

References 10 publications

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“…Capillary voltage was kept at 4.3 kv and capillary temperature set at 285°C. Collision induced energy range varied from 20-30 (Mirjalili et al, 2013).…”

Section: Epi-ms/ms Of Withania Coagulansmentioning

confidence: 99%

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2021

Pak. J. Pharm. Sci

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Herbal medicines are gaining importance due to more advantages and less toxic effects. Withania coagulans is natural plant that possesses multiple activities. Its main constituents are withaferin and withanolide. The purpose of present study is to identify main constituent of Withania coagulans and preparation of extract loaded micro emulsions. Withania coagulans fruit extract in methanol/chloroform (1:1) was collected in semisolid form and LCMS was done to identify active compound, and then micro emulsions were prepared using Tween 80: Transcutol (1:1) Frankincense oil, and water to enhance its stability for topical application. Five formulations were prepared by Pseudo ternary phase diagram and evaluated for pH, conductivity, viscosity, drug contents, particle size analysis, and polydispersity. Withania coagulans extract was evaluated for anti-bacterial activity against (Staphylococcus aureus, E. coli, and S. typhi) and antifungal activity against (Candida albicans and Aspergillus niger). Anti-inflammatory activity was checked for both extract and Extract based micro emulsion. Among all five formulations F5 shows best physiochemical properties with small globule size, good stability and high anti-inflammatory activity. Based on these results it was concluded that Withania coagulans extract loaded micro emulsions can be used for topical application with promising anti-inflammatory activities. Data for in-vivo studies for checking the topical effect of Withania coagulans is provided elsewhere.

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“…Capillary voltage was kept at 4.3 kv and capillary temperature set at 285°C. Collision induced energy range varied from 20-30 (Mirjalili et al, 2013).…”

Section: Epi-ms/ms Of Withania Coagulansmentioning

confidence: 99%

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2021

Pak. J. Pharm. Sci

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“…These compounds are specific for Solanaceae family, in particular, for the genus Withania, and thus they were used as marker compounds 13 . WA was estimated from WS and different extracts there of as well as from herbal formulations contained WS, as one of the ingredients, using TLC and HPLC [14][15][16][17][18][19][20][21][22][23][24][25][26][27][28][29][30][31] , HPLC-MS/MS 32 , UHPLC-MS/MS 33…”

Section: Fig 1: Chemical Structure Of Wamentioning

confidence: 99%

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2020

IJPSR

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Ashwagandhadi lehya is an Ayurveda formulation, official in Ayurvedic Formulary of India and used as aphrodisiac, tonic, rejuvenating agents etc. It is one of the major components of Ashwagandhadi lehya is Withania somnifera which contain Withaferin-A. The study was undertaken to develop an analytical method using HPLC to estimate Withaferin-A, being a marker from, methanolic extract of Ashwagandhadi lehya as one of the quality parameters. Method development involved simple sample preparation without any purification, as well as isocratic separation of Withaferin-A. The method was subjected to validation involving establishing linearity, precision, accuracy, specificity, and robustness.Optimized mobile phase was acetonitrile: water (45: 55 v/v) contained 1% v/v glacial acetic acid. The column used was Phenomenex (250 × 4.6 mm, 5 µm) with flow rate set at 1 ml/min and chromatogram was recorded at 216 nm. The studies showed that peak corresponding to Withaferin-A within concentration range of 1-75 µg/ml was found to be correlated with respective concentration with correlation coefficient (r 2) 0.99802 ± 0.0004. The developed analytical method was found precise, while accuracy was established using standard addition method. The average recovery was 95.59%, when determined at three different spiking levels (50%, 100% & 150%), in the preanalyzed sample solution. LOD was found to be 0.3 µg/ml and LOQ was 0.1 µg/ml. Robustness study suggested that increase in acetonitrile proportion, increased R T as well as increase in flow rate negatively impacted up on R T. The content of Withaferin-A was estimated as 0.03 ± 0.001 mg % w/w. The results obtained from the studies showed that method was found to be accurate, precise and validated for estimation of Withaferin-A from Ashwagandhadi lehya. INTRODUCTION: Ashwagandhadi lehya (AL) is a polyherbal formulation, official in Ayurvedic formulary of India. AL is indicated in the treatment of piles, psychosis, as tonic, rejuvenating agents and in treatment of disorders related to male reproductive system. One of the major ingredients of AL is dried powdered roots of Withania sominfera (WS).

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“…The literature on standardization of WS shows reports of high-performance liquid chromatography (HPLC), − high-performance thin-layer chromatography, − and liquid chromatography–mass spectrometry (LC–MS)-based − methodologies with the single or multiple bioactive or chemical markers. In these, withanolides and withanosides have been studied separately or together.…”

Section: Introductionmentioning

confidence: 99%

Investigating 11 Withanosides and Withanolides by UHPLC–PDA and Mass Fragmentation Studies from Ashwagandha (Withania somnifera)

et al. 2020

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Withania somnifera (WS), also known as ashwagandha or Indian ginseng, is known for its pharmacological significance in neurodegenerative diseases, stress, cancer, immunomodulatory, and antiviral activity. In this study, the WS extract (WSE) from the root was subjected to ultrahigh-performance liquid chromatography with photodiode array detection (UHPLC− PDA) analysis to separate 11 withanoside and withanolide compounds. The quantification validation was carried out as per ICHQ2R1 guidelines in a single methodology. The calibration curves were linear (r 2 > 0.99) for all 11 compounds within the tested concentration ranges. The limits of detection and quantification were in the range of 0.213−0.362 and 0.646−1.098 μg/mL, respectively. The results were precise (relative standard deviation, <5.0%) and accurate (relative error, 0.01−0.76). All compounds showed good recoveries of 84.77−100.11%. For the first time, withanoside VII, 27-hydroxywithanone, dihydrowithaferin A, and viscosalactone B were quantified and validated along with bioactive compounds withanoside IV, withanoside V, withaferin A, 12deoxywithastramonolide, withanolide A, withanone, and withanolide B simultaneously in WS. This UHPLC−PDA method has practical adaptability for ashwagandha raw material, extract, and product manufacturers, along with basic and applied science researchers. The method has been developed on UHPLC for routine analysis. The 11 withanosides and withanolides were confirmed using the fragmentation pattern obtained by the combined use of electrospray ionization and collision-induced dissociation in triplequadrupole tandem mass spectrometry (TQ−MS/MS) in the WSE.

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LC-MS/MS method for the quantification of withaferin-A in plant extracts ofWithaniaspp.

Cited by 7 publications

References 10 publications

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Investigating 11 Withanosides and Withanolides by UHPLC–PDA and Mass Fragmentation Studies from Ashwagandha (Withania somnifera)

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