title: Screening for in vitro metabolites of kakkalide and irisolidone in human and rat intestinal bacteria by ultra-high performance liquid chromatography/quadrupole time-of-flight mass spectrometry.
abstract: Kakkalide and irisolidone, the main isoflavones of Flos Puerariae, exhibit a wide spectrum of bioactivities. Intestinal bacteria biotransformation plays an important role in the metabolic pathways of flavones, and is directly related to the bioactivities of the prodrugs after oral administration. To the best of our knowledge, the metabolic pathways of kakkalide and irisolidone in vitro have not been comprehensively studied yet. This paper describes the strategy using ultra-high performance liquid chromatography/quadrupole time-of-flight mass spectrometry (UHPLC/Q-TOF MS) for the rapid analysis of the metabolic profiles of kakkalide and irisolidone after incubated with human and rat intestinal bacteria. Bacteria incubated samples were prepared and analyzed after incubated under anaerobic conditions for 48 h. A total of 17 metabolites, including parent compounds, were detected in human and rat intestinal bacteria incubated samples. The results obtained indicate that hydrolysis, dehydroxylation, demethoxylation, demethylation, hydroxylation, decarbonylation, and reduction were the detected metabolic pathways of kakkalide and irisolidone in vitro. The conversion rate of irisolidone in human and rat bacteria was 8.57% and 6.51%, respectively. Biochanin A was the relatively main metabolite of irisolidone, and the content of biochanin A in human and rat bacteria was 3.68% and 4.25%, respectively. The conversion rate of kakkalide in human and rat bacteria was 99.92% and 98.58%, respectively. Irisolidone was the main metabolite of kakkalide, and the content of irisolidone in human and rat bacteria was 89.58% and 89.38%, respectively. This work not only provides the evidence of kakkalide and irisolidone metabolites in vivo, but also demonstrates a simple, fast, sensitive, and inexpensive method for identification of metabolites of other compounds transformed by intestinal bacteria.
authors.affiliation: Department of Traditional Chinese Medicine, Shenyang Pharmaceutical University, 103 Wenhua Road, Shenyang 110016, China.
authors.forename: Guozhe
authors.initials: G
authors.lastname: Zhang
authors.affiliation: Department of Materials Engineering, University of British Columbia, Frank Forward Building, 6350 Stores Road, Vancouver, BC V6T 1Z4, Canada.
authors.forename: Tianxing
authors.initials: T
authors.lastname: Gong
authors.affiliation: Department of Traditional Chinese Medicine, Shenyang Pharmaceutical University, 103 Wenhua Road, Shenyang 110016, China.
authors.forename: Yoshihiro
authors.initials: Y
authors.lastname: Kano
authors.affiliation: Department of Traditional Chinese Medicine, Shenyang Pharmaceutical University, 103 Wenhua Road, Shenyang 110016, China. Electronic address: yuandan_kampo@163.com.
authors.forename: Dan
authors.initials: D
authors.lastname: Yuan
mesh_terms: D000328:Adult; D000818:Animals; D001419:Bacteria / Q000378:metabolism*; D001711:Biotransformation; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D005243:Feces / Q000382:microbiology; D005419:Flavonoids / Q000737:chemistry* / Q000378:metabolism; D035264:Flowers / Q000737:chemistry / Q000378:metabolism; D006027:Glycosides / Q000737:chemistry* / Q000378:metabolism; D006801:Humans; D007413:Intestinal Mucosa / Q000378:metabolism; D007422:Intestines / Q000382:microbiology*; D007529:Isoflavones / Q000737:chemistry* / Q000378:metabolism; D008297:Male; D013058:Mass Spectrometry / Q000379:methods*; D015394:Molecular Structure; D010936:Plant Extracts / Q000737:chemistry / Q000378:metabolism*; D029908:Pueraria / Q000737:chemistry / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley
chemical_list: D005419:Flavonoids; D006027:Glycosides; D007529:Isoflavones; D010936:Plant Extracts; C423902:irisolidone; C108698:kakkalide
keywords: BiA; Biotransformation; CLogP; ESI; Glc; Intestinal bacteria; Ir; Irisolidone; KA; Kakkalide; Metabolic profile; Te; UHPLC/Q-TOF MS; Xyl; biochanin A; calculated 1-octanol/water partition coefficient; electro spray ionization; glucose; irisolidone; kakkalide; tectorigenin; ultra-high performance liquid chromatography/quadrupole time-of-flight mass spectrometry; xylose