  Citation: Ansari, M.H.R.; Khan, W.; Parveen, R.; Saher, S.; Ahmad, S. Pharmacokinetic, Metabolomic, and Stability Assessment of Ganoderic Acid H Based Triterpenoid Enriched Fraction of Ganoderma lucidum P. Karst. Metabolites 2022, 12, 97. https://doi.org/10.3390/ metabo12020097 Academic Editors: Alba Rodríguez-Nogales, Maria Elena Rodriguez-Cabezas and Leonardo Tenori Received: 30 September 2021 Accepted: 9 December 2021 Published: 21 January 2022 Publisher’s Note: MDPI stays neutral with regard to jurisdictional claims in published maps and institutional affil- iations. Copyright: © 2022 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https:// creativecommons.org/licenses/by/ 4.0/). metabolites H OH OH Article Pharmacokinetic, Metabolomic, and Stability Assessment of Ganoderic Acid H Based Triterpenoid Enriched Fraction of Ganoderma lucidum P. Karst Mohd Hafizur Rehman Ansari 1 , Washim Khan 1 , Rabea Parveen 1,2 , Sadia Saher 3 and Sayeed Ahmad 1, * 1 Bioactive Natural Product Laboratory, School of Pharmaceutical Education and Research, Jamia Hamdard, New Delhi 110062, India; rehman.ansari1988@gmail.com (M.H.R.A.); khan.washim78@gmail.com (W.K.); rabea_nd62@yahoo.co.in (R.P.) 2 Human Genetics Laboratory, Department of Bioscience, Jamia Millia Islamia, New Delhi 110025, India 3 Viral Research Development Laboratory, Department of Microbiology, J.N.M.C, A.M.U, Aligarh 202002, India; sadia.micro@gmail.com * Correspondence: sahmad_jh@yahoo.co.in Abstract: Ganoderma lucidum P. karst is an edible fungus that is used in traditional medicine and contains triterpenoids as the major phytoconstituents. Ganoderic acids are the most abundant triter- penoids that showed pharmacological activity. As Indian varieties contain ganoderic acid H (GA-H), we aimed to prepare GA-H-based triterpenoid enriched fraction (TEF) and evaluated its pharmacoki- netics, metabolomics, and stability analysis. A high-performance liquid chromatography (HPLC) method was developed to quantify GA-H in TEF and rat plasma. Based on GA-H content, a stability assessment and pharmacokinetic study of TEF were also performed. After its oral administration to rats, TEF’s the metabolic pattern recognition was performed through ultra-performance liquid chromatography mass spectroscopy (UPLC–MS). The developed HPLC method was found to be simple, sensitive, precise (<15%), and accurate (>90% recovery) for the quantification of GA-H. Phar- macokinetic analysis showed that GA-H reached its maximum plasma concentration (C max 2509.9 ng/mL) within two hours and sustained quantifiable amount up to 12 h with a low elimination rate (K el ) 0.05 L/h. TEF contained ten bioavailable constituents. The prepared TEF was found to be stable for up to one year at room temperature. The prepared TEF, enriched with ganoderic acid, is stable, contains bioavailable constituents, and can be explored as phytopharmaceuticals for different pharmacological properties. Highlights: (1). Preparation of triterpenoid enriched fraction (TEF) from Ganoderma lucidum. (2). Major triterpenoid in TEF is ganoderic acid H (GA-H). (3). TEF contains several bioavailable phytoconstituents. (4). TEF (considering only GA-H) is stable for up to one year at room temperature. (5). GA-H is rapidly absorbed and has high systemic exposure. Keywords: triterpenoid; ganoderic acid H; quantitative analysis; UPLC–MS; stability 1. Introduction The intake of dietary supplements has been increased alarmingly due to several health benefits and trend toward healthy lifestyles. There are numerous herbs and living or- ganisms used as dietary supplements. Fungi are the most widely distributed organisms on earth and are of great medicinal importance. Ganoderma lucidum, a fungus known as ‘Mushroom’ in India or ‘Lingzhi’ in China or ‘Reishi’ in Japan, is a well-known medicinal mushroom and has a great future as a dietary supplement. Over 2000 years, G. lucidum has been used in several traditional systems as rejuvenation of health by providing vital en- ergy, strengthening the immune system, anti-aging, and strengthening body resistance [1]. Nowadays, it is widely used as a dietary supplement in the United States and in European countries. It is enriched with different classes of secondary metabolites, such as triter- penoids, polysaccharides, peptides, alkaloids, steroids, flavonoids, iridoids, and lignans, Metabolites 2022, 12, 97. https://doi.org/10.3390/metabo12020097 https://www.mdpi.com/journal/metabolites