Simultaneous Quantification of Antioxidant Compounds in Phellinus igniarius Using Ultra Performance Liquid Chromatography-Photodiode Array Detection-Electrospray Ionization Tandem Mass Spectrometry

Abstract

Natural antioxidants are widely used in the life sciences. Phellinus igniarius is a historically used natural antioxidant containing a variety of active compounds. Phenols, particularly Inoscavin A and Hypholomine B, are found in the high concentrations. Better quantitative methods are needed to perform quality control in order to support further research of this mushroom. An ultra-performance liquid chromatography method coupled to photodiode-array detection and an electrospray ionization tandem mass spectrometry method (UPLC-PAD-MS) was developed to simultaneously quantify Inoscavin A and Hypholomine B levels in the medicinal fungus Phellinus igniarius. The two compounds were quantified using UPLC-PAD and UPLC-MS. The methods were accurate (mean accuracy for spiked matrix ranged from 101.5% to 105.8%), sensitive (limit of detection ranged from 0.28 to 1.14 mg L-1) and precise (the relative standard deviations ranged from 0.13 to 2.8%). Inoscavin A and Hypholomine B were purified using high-speed counter-current chromatography (HSCCC), structural evaluated to meet the request of standard substances. UPLC separation was performed on a reversed-phase C18 column using gradient elution with acetonitrile and 0.1% formic acid over 10 min. The developed method was successfully applied to determine Inoscavin A and Hypholomine B in twelve Phellinus igniarius samples of different origins and the results showed that it was suitable for the analysis of these active components in Phellinus igniarius samples.

Anti-Gout Effects of the Medicinal Fungus Phellinus igniarius in Hyperuricaemia and Acute Gouty Arthritis Rat Models

“…Furthermore, to elucidate the main active components in WPP and CPP better, UPLC-ESI-qTOF-MS, which is characterized by fast separation, high sensitivity, and accurate relative molecular mass calculation ( Ye et al, 2021 ), was used for identification. Surprisingly, similar active ingredients, such as protocatechuic aldehyde, hispidin, davallialactone, phelligridimer A, hypholomine B and inoscavin A, were identified in both WPP and CPP by UPLC-ESI-qTOF-MS. As members of the polyphenol family, the antioxidant and anti-inflammatory activities of protocatechuic aldehyde ( Chang et al, 2011 ), hispidin ( Jin et al, 2021 ), davallialactone ( Lee et al, 2008 ; Lee et al, 2011 ), phelligridimer A ( Wang et al, 2005b ), hypholomine B ( Shou et al, 2016 ) and inoscavin A ( Lee et al, 2006 ; Shou et al, 2016 ) have been widely reported. A derivative of protocatechuic aldehyde effectively inhibited XO activity and reduced serum uric acid levels in hyperuricaemia mice ( Lü et al, 2013 ).…”

A Combined Phytochemistry and Network Pharmacology Approach to Reveal the Potential Antitumor Effective Substances and Mechanism of Phellinus igniarius

“…Phellinus igniarius ( P. igniarius ), a well-known mushroom belonging to the genus Phellinus in the polyporaceae family, is a physiologically functional food and exemplary source of natural medicine that has been widely used in China, Japan, Korea, and other countries ( Ma et al, 2016 ). P. igniarius possesses high anti-inflammatory, antioxidant and antitumor biological activities due to its accumulation of various secondary metabolites, including polysaccharides, flavonoids, polyphenols, steroids and organic acids ( Dong et al, 2015 ; Shou et al, 2016 ). Notably, recent studies have creatively focused on the potential functions of P. igniarius extracts and their constituent compounds in the prevention and treatment of cancer ( Zhou et al, 2014 ; Sangdee et al, 2017 ).…”

Development of RP HPLC‐PDA method for simultaneous quantitative analysis of Inoscavin A and Meshimakobnol A and application on some Phellinus mushroom species

“…The other study by Dan Shou et al. ( 2016 ) (Shou et al., 2016 ) quantitated Inoscavin A without Meshimakobnol A which could be a differentiation profiling between Phellinus and other mushrooms. Besides, our experimental data shows that Inoscavin A is one of the major pigments in Phellinus species in Vietnam.…”

“…While, the chromatographic observations of the above study also revealed differences between wild, cultivated, and commercial mushrooms not only in the composition of Meshimakobnol A but also in Inoscavin A which could be a composition to distinguish among wild and cultivated mushrooms. The other study byDan Shou et al (2016)(Shou et al, 2016) quantitated Inoscavin A without Meshimakobnol A which could be a differentiation profiling between Phellinus and other mushrooms. Besides, our experimental data shows that Inoscavin A is one of the major pigments in Phellinus species in Vietnam.…”