Isolation of Mirificin and Other Bioactive Isoflavone Glycosides from the Kudzu Root Lyophilisate Using Centrifugal Partition and Flash Chromatographic Techniques

Abstract

Pueraria lobata (Willd.) Ohwi is a legume taxon native to Southeast Asia and widely used in traditional medicine systems of that region. The therapeutic applications of the underground parts of this species (known as kudzu root) are related to its high content of isoflavones, mainly the characteristic C-glycoside derivatives. Within this group, the most explored compound both phytochemically and pharmacologically is puerarin. However, current scientific findings document important anti-biodegenerative effects for some of the minor isoflavones from kudzu roots. Therefore, the main objective of the study was to develop an original preparative method that allowed the efficient isolation of closely related hydrophilic daidzein C-glycosides, including mirificin, from vacuum-dried aqueous-ethanolic extracts of kudzu roots. For this purpose, the combined centrifugal partition (CPC) and flash chromatographic (FC) techniques were elaborated and used. The optimized biphasic solvent system in CPC, with ethyl acetate, ethanol, water, and 0.5% (V/V) acetic acid as a mobile phase modifier, enabled the purification and separation of the polar fraction containing bioactive isoflavones and ultimately the isolation of mirificin, 3′-hydroxy- and 3′-methoxypuerarin, puerarin, and daidzin using FC. The identity of isoflavones was confirmed using spectroscopic (UV absorption and nuclear magnetic resonance) and mass spectrometric methods. The determined purity of isolated mirificin was 63%.

Optimization of the Zhou Tian Formula extraction technology based on AHP‐CRITIC method and analysis of transfer efficiency of key components based on HPLC fingerprinting

“…In addition, the peak area data from the granule fingerprint showed that the peak area of puerarin and mirificin was the largest among all the peak areas, indicating their remarkable significance in terms of quality control. A few studies have shown that calycosin is a component of Astragali Radix, and puerarin and mirificin are important components of Pueraria Lobata 25,32 . These findings underscore the necessity for stringent quality control measures for fried Astragali Radix and Pueraria Lobata to ensure the stability of ZTF granules.…”

“…31 Components with VIP values greater than 1 played a crucial role in of Pueraria Lobata. 25,32 These findings underscore the necessity for stringent quality control measures for fried Astragali Radix and Pueraria Lobata to ensure the stability of ZTF granules.…”

“…), with its peak area ranking second among all peak areas. 25 Given the pharmacological mechanisms of action of these components and their substantial quantity, they were utilized as index components for evaluation. The weight coefficients of these indicators were calculated using the AHP-CRITIC method.…”

Approaches, Strategies and Procedures for Identifying Anti-Inflammatory Drug Lead Molecules from Natural Products

“…Ohwi. by combined centrifugal partition (CPC) and FC methods [ 171 ]. The current advancements include fully automated flash chromatography equipment equipped with robotic fraction collectors and online detection units, significantly enhancing the efficiency of separating, isolating, and purifying constituent compounds within a complex mixture of crude extract and identification [ 172 ].…”

Advances in Extraction, Purification, and Analysis Techniques of the Main Components of Kudzu Root: A Comprehensive Review

“…Kudzu root (Pueraria lobate (Willd.) Ohwi, KR), a legume taxon indigenous to Southeast Asia, boasts a rich variety of species and can be found across China [ 1 ], including in Hebei, Guangxi, Anhui, Liaoning, and Shandong. In fact, it has become one of the most prevalent twining shrub plants in urban forests.…”