Kapp Karmen, Gafriller Johannes, Kirchweger Benjamin, Rollinger Judith M, Grienke Ulrike
Drug Research Program, Division of Pharmaceutical Biosciences, Faculty of Pharmacy, University of Helsinki, 00014 Helsinki, Finland; Division of Pharmacognosy, Department of Pharmaceutical Sciences, Faculty of Life Sciences, University of Vienna, Josef-Holaubek-Platz 2, 1090 Vienna, Austria.
Division of Pharmacognosy, Department of Pharmaceutical Sciences, Faculty of Life Sciences, University of Vienna, Josef-Holaubek-Platz 2, 1090 Vienna, Austria; Vienna Doctoral School of Pharmaceutical, Nutritional and Sport Sciences, University of Vienna, Josef-Holaubek-Platz 2, 1090 Vienna, Austria.
J Chromatogr A. 2025 Sep 13;1758:466197. doi: 10.1016/j.chroma.2025.466197. Epub 2025 Jul 5.
Inonotus obliquus is a fungal pathogen that parasitizes the trunks of broadleaved tree species. The sclerotia of this fungus, so-called chaga, show a broad range of traditional and medicinal uses. The increasing market for chaga supplements, promoted for their antioxidant, anti-inflammatory, and antidiabetic properties, highlights the need for standardized identity and quality control methodologies. In this study, the first supercritical fluid-based protocol for the extraction, isolation, identification, and quantitation of six characteristic chaga constituents [lanosterol (1), ergosterol (2), inotodiol (3), trametenolic acid B (4), betulin (5) and inonotsutriol A (6)] was developed and validated. Semi-preparative supercritical fluid chromatography was utilized, to obtain lanosterol (1), inotodiol (3) and inonotsutriol A (6). Ultra-high-performance supercritical fluid chromatography allowed for a separation of the standard compounds within 10 min. Good results regarding linearity (R ≥ 0.99) were obtained for photodiode array detection, with a lowest detection limit of 0.424 μg/mL. A 20-minute exhaustive SFE method was applied to 14 commercial chaga samples, yielding extracts ranging from 0.17 % to 0.58 %. The qualitative composition of the extracts was consistent, with inotodiol (3), trametenolic acid B (4), and lanosterol (1) as the most abundant constituents. Notably, inonotsutriol A (6) was quantified in commercial chaga samples for the first time. The use of supercritical fluid technology enabled a fast, selective, and environmentally friendly extraction, isolation, and analysis of compounds. This protocol offers a sustainable alternative to traditional chromatographic methods and could serve as a valuable tool for the authenticity and quality control of chaga raw materials and supplements.
桦褐孔菌是一种寄生于阔叶树种树干的真菌病原体。这种真菌的菌核，即所谓的桦褐孔菌，具有广泛的传统和药用用途。桦褐孔菌补充剂市场不断增长，因其具有抗氧化、抗炎和抗糖尿病特性而受到推广，这凸显了对标准化身份识别和质量控制方法的需求。在本研究中，开发并验证了首个基于超临界流体的方案，用于提取、分离、鉴定和定量六种桦褐孔菌特征成分[羊毛甾醇(1)、麦角甾醇(2)、氧化白腐菌醇(3)、栓菌酸B(4)、桦木醇(5)和桦褐孔菌醇A(6)]。采用半制备超临界流体色谱法获得羊毛甾醇(1)、氧化白腐菌醇(3)和桦褐孔菌醇A(6)。超高效超临界流体色谱法可在10分钟内分离标准化合物。二极管阵列检测的线性关系良好(R≥0.99)，最低检测限为0.424μg/mL。采用20分钟的全量超临界流体萃取法对14个市售桦褐孔菌样品进行处理，提取物含量在0.17%至0.58%之间。提取物的定性组成一致，氧化白腐菌醇(3)、栓菌酸B(4)和羊毛甾醇(1)为最主要成分。值得注意的是，首次在市售桦褐孔菌样品中对桦褐孔菌醇A(6)进行了定量。超临界流体技术的应用实现了化合物的快速、选择性和环境友好型提取、分离和分析。该方案为传统色谱方法提供了一种可持续的替代方法，可作为桦褐孔菌原材料和补充剂真伪鉴定和质量控制的有价值工具。
