A UPLC-MS/MS Based Rapid, Sensitive, and Non-Enzymatic Methodology for Quantitation of Dietary Isoflavones in Biological Fluids

Faraz Rashid; Sudeep Ghimire; Ashutosh K. Mangalam; Shailendra Giri

doi:10.3390/molecules28186729

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A UPLC-MS/MS Based Rapid, Sensitive, and Non-Enzymatic Methodology for Quantitation of Dietary Isoflavones in Biological Fluids

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A UPLC-MS/MS Based Rapid, Sensitive, and Non-Enzymatic Methodology for Quantitation of Dietary Isoflavones in Biological Fluids

Faraz Rashid, Sudeep Ghimire, Ashutosh K. Mangalam and Shailendra Giri

Molecules (Basel, Switzerland), Vol.28(18), 6729

09/21/2023

DOI: 10.3390/molecules28186729

PMCID: PMC10534480

PMID: 37764503

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Abstract

Dietary isoflavones, a type of phytoestrogens, have gained importance owing to their health-promoting benefits. However, the beneficial effects of isoflavones are mediated by smaller metabolites produced with the help of gut bacteria that are known to metabolize these phytoestrogenic compounds into Daidzein and Genistein and biologically active molecules such as S-Equol. Identifying and measuring these phytoestrogens and their metabolites is an important step towards understanding the significance of diet and gut microbiota in human health and diseases. We have overcome the reported difficulties in quantitation of these isoflavones and developed a simplified, sensitive, non-enzymatic, and sulfatases-free extraction methodology. We have subsequently used this method to quantify these metabolites in the urine of mice using UPLC-MS/MS. The extraction and quantitation method was validated for precision, linearity, accuracy, recoveries, limit of detection (LOD), and limit of quantification (LOQ). Linear calibration curves for Daidzein, Genistein, and S-Equol were set up by performing linear regression analysis and checked using the correlation coefficient (r2 > 0.995). LOQs for Daidzein, Genistein, and S-Equol were 2, 4, and 2 ng/mL, respectively. This UPLC-MS/MS swift method is suitable for quantifying isoflavones and the microbial-derived metabolite S-Equol in mice urine and is particularly useful for large numbers of samples.

Details

Title: Subtitle: A UPLC-MS/MS Based Rapid, Sensitive, and Non-Enzymatic Methodology for Quantitation of Dietary Isoflavones in Biological Fluids
Creators: Faraz Rashid - Henry Ford Health System
Sudeep Ghimire - University of Iowa
Ashutosh K. Mangalam - University of Iowa
Shailendra Giri - Henry Ford Health System
Resource Type: Journal article
Publication Details: Molecules (Basel, Switzerland), Vol.28(18), 6729
DOI: 10.3390/molecules28186729
PMID: 37764503
PMCID: PMC10534480
NLM abbreviation: Molecules
ISSN: 1420-3049
eISSN: 1420-3049
Grant note: name: HFH, award: A31001; DOI: 10.13039/100000009, name: National Institutes of Health/NIAID, award: 1RO1AI137075; name: Veteran Affairs Merit Award, award: 1I01CX002212; DOI: 10.13039/100000890, name: National Multiple Sclerosis Society, award: RG-2111-38733; name: National Institutes of Health, award: NS112727, AI144004; name: Henry Ford Hospital Internal support, award: A10270, A30967; name: NIH Interdisciplinary Immunology training grant, award: T32AI007260
Language: English
Date published: 09/21/2023
Academic Unit: Pathology; Iowa Neuroscience Institute
Record Identifier: 9984472654802771

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