Pharmacokinetic Study of Withanosides and Withanolides from Withania somnifera Using Ultra-High Performance Liquid Chromatography-Tandem Mass Spectrometry (UHPLC-MS/MS)

Abstract

Withania somnifera is a traditional Indian herb described under the ‘Rasayana’ class in Ayurveda, which gained immense popularity as a dietary supplement in the USA, Europe, Asia, and the Indian domestic market. Despite enormous research on the pharmacological effect of withanosides and withanolides, bioanalytical method development and pharmacokinetics remained challenging and unexplored for these constituents due to isomeric and isobaric characteristics. In current research work, molecular descriptors, pharmacokinetic, and toxicity prediction (ADMET) of these constituents were performed using Molinspiration and admetSAR tools. A rapid, selective, and reproducible bioanalytical method was developed and validated for seven withanosides and withanolides as per USFDA/EMA guidelines, further applied to determine pharmacokinetic parameters of Withania somnifera root extract (WSE) constituents in male Sprague Dawley rats at a dose of 500 mg/kg. Additionally, an ex vivo permeability study was carried out to explore the absorption pattern of withanosides and withanolides from the intestinal lumen. In silico, ADMET revealed oral bioavailability of withanosides and withanolides following Lipinski’s rules of five with significant absorption from the gastrointestinal tract and the ability to cross the blood-brain barrier. Upon oral administration of WSE, Cmax was found to be 13.833 ± 3.727, 124.415 ± 64.932, 57.536 ± 7.523, and 7.283 ± 3.341 ng/mL for withanoside IV, withaferin A, 12-Deoxy-withastramonolide, and withanolide A, respectively, with Tmax of 0.750 ± 0.000, 0.250 ± 0.000, 0.291 ± 0.102, and 0.333 ± 0.129 h. Moreover, at a given dose, withanoside V, withanolide B, and withanone were detected in plasma; however, the concentration of these constituents was found below LLOQ. Thus, these four major withanoside and withanolides were quantified in plasma supported by ex vivo permeation data exhibiting a time-dependent absorption of withanosides and withanolides across the intestinal barrier. These composite findings provide insights to design a clinical trial of WSE as a potent nutraceutical.

In vitro effect of Withania somnifera, AYUSH-64, and remdesivir on the activity of CYP-450 enzymes: Implications for possible herb−drug interactions in the management of COVID-19

“…Both the test drugs might follow the BCS Class-I nature of one-compartment distribution ( Patil et al, 2014 ), and it is expected to achieve less than <100 μg/ml concentration for the extracts and <100 μM/ml for the individual plant phytoconstituents. It has been verified in in vivo experiments ( Patil et al, 2013 ; Modi et al, 2022 ). Therefore, the K i value can be calculated with the aforementioned formula.…”

“…Briefly, the Ayurvedic drug properties (rasa, guna, virya, vipaka, and prabhava) are helpful in deciding to use the test drugs and their putative PK-PD approaches [Gogte, 2000;Mishra et al, (Patil et al, 2014), and it is expected to achieve less than <100 μg/ml concentration for the extracts and <100 μM/ml for the individual plant phytoconstituents. It has been verified in in vivo experiments (Patil et al, 2013;Modi et al, 2022). Therefore, the K i value can be calculated with the aforementioned formula.…”

Evaluating anticancer properties of Withaferin A—a potent phytochemical

“…Furthermore, Withaferin A and 1,2deoxywithastramonolide constituents were detectable in plasma up till 10 h which depicted possibility of lower elimination rate for these constituents (Modi et al, 2022).…”

“…The study found reduction in LDL and overall cholesterol levels, increased strength of muscle activity, and normal organ function levels (Raut et al, 2012). Modi et al (2022) performed an ADMET study on Withania somnifera extracts and assessed genotoxicity and mutagenicity at 500 g/kg dose. The ADMET study revealed none of the seven constituents of Withania somnifera extract, including Withaferin A, to be hepatotoxic, mutagenic, or carcinogenic (Modi et al, 2022).…”

“…A pharmacokinetic study was conducted by Modi et al (2022) , by orally administering 500 mg/kg dose of Withania somnifera root extract to male Sprague Dawley rats at 500 mg/kg and determined the absorption patterns of various withanolides, including Withaferin A ( Modi et al, 2022 ). Rapid absorption of withanolide and withanoside constituents from the stomach was indicated by the results ( Modi et al, 2022 ).…”