A Rapid and Reliable Spectrofluorimetric Method to Measure the Urinary Lactulose/Mannitol Ratio for Dysbiosis Assessment

Gut microbiota plays a crucial role in human health homeostasis, and the result of its alteration, known as dysbiosis, leads to several pathologies (e.g., inflammatory bowel disease, metabolic syndrome, and Crohn's disease). Traditional methods used to assess dysbiosis include the dual sugar ab...

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Bibliographic Details
Published in:Biomedicines 2024-07, Vol.12 (7), p.1557
Main Authors: Marino Cerrato, Lorenzo, Schiano, Elisabetta, Iannuzzo, Fortuna, Tenore, Gian Carlo, Summa, Vincenzo, Daglia, Maria, Novellino, Ettore, Stornaiuolo, Mariano
Format: Article
Language:English
Subjects:
Automation
Calibration
Carbohydrates
Crohn's disease
Dehydrogenases
Dysbacteriosis
dysbiosis
Environmental impact
Enzymes
Food
Glucose
High-throughput screening
Homeostasis
Inflammatory bowel diseases
Intestinal microflora
Lactose
Lactulose
LMR
Mannitol
Mass spectroscopy
Metabolic syndrome
Microbiota
NADPH-diaphorase
Permeability
spectrofluorimetry
Storage conditions
Urine
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Summary:Gut microbiota plays a crucial role in human health homeostasis, and the result of its alteration, known as dysbiosis, leads to several pathologies (e.g., inflammatory bowel disease, metabolic syndrome, and Crohn's disease). Traditional methods used to assess dysbiosis include the dual sugar absorption test and the urinary lactulose/mannitol ratio (LMR) measurement using mass spectrometry. Despite its precision, this approach is costly and requires specialized equipment. Hence, we developed a rapid and reliable spectrofluorimetric method for measuring LMR in urine, offering a more accessible alternative. This spectrofluorimetric assay quantifies the fluorescence of nicotinamide adenine dinucleotide (NADH) and nicotinamide adenine dinucleotide phosphate (NADPH) produced during the enzymatic oxidation of mannitol and lactulose, respectively. The assay requires 100 µL of urine samples and detects LMR values lower (eubiosis) and higher (dysbiosis) than 0.05, ultimately being amenable to high-throughput screening and automatization, making it practical for clinical and research settings. A validation of the method demonstrated its high precision, accuracy, and robustness. Additionally, this study confirmed analyte stability under various storage conditions, ensuring reliable results even with delayed analysis. Overall, this spectrofluorimetric technique reduces costs, time, and the environmental impact associated with traditional mass spectrometry methods, making it a viable option for widespread use in the assessment of dysbiosis.
ISSN:2227-9059
2227-9059
DOI:10.3390/biomedicines12071557