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Simple and sensitive multi-sugar-probe gut permeability test by high-performance liquid chromatography with
D R Rooyakkers1, H M van Eijk, N E Deutz
1Department of Surgery, University of Limburg, Maastricht, The Netherlands.
Journal of Chromatography. A
|April 12, 1996
Summary
A new method using FMOC-hydrazine significantly enhances the sensitivity of gut permeability testing by accurately measuring sugar probes in urine. This allows for lower, safer doses of lactulose, reducing the risk of diarrhea and improving test reliability.
Area of Science:
- Biochemistry
- Analytical Chemistry
- Gastroenterology
Background:
- Gut permeability is assessed using non-metabolic sugar probes, but traditional methods lack sensitivity, requiring high doses that can cause side effects like diarrhea.
- Existing assays often struggle with sensitivity, necessitating large ingestions of probes such as lactulose, which can lead to adverse gastrointestinal effects and impact test accuracy.
Purpose of the Study:
- To develop a highly sensitive method for measuring reducing sugar probes in urine using 9-fluorenylethyl chloroformate hydrazine (FMOC-hydrazine).
- To reduce the required dose of orally administered probes, thereby minimizing patient discomfort and potential test interference.
Main Methods:
- Utilized FMOC-hydrazine to derivatize reducing sugars (lactulose, rhamnose, xylose, 3-O-methyl-D-glucose) for sensitive detection in aqueous samples.
- Validated the method's linearity, limit of detection, precision (coefficient of variation), and recovery in urine samples.
- Compared urinary probe ratios in human volunteers with literature values and assessed the optimal time for sample collection.
Main Results:
- The FMOC-hydrazine method demonstrated excellent linearity (R2 > 0.9997) and very low limits of detection (0.05 pmol for lactulose, 0.01 pmol for others).
- Sensitivity improvements allowed a reduction in lactulose intake from 6-10 g to 1.5 g, significantly minimizing diarrhea risk.
- High recovery rates (>100%) and low coefficients of variation (<3%) confirmed method precision and accuracy. Urinary lactulose-rhamnose ratios in volunteers were comparable to literature values after just 3 hours.
Conclusions:
- The developed FMOC-hydrazine based assay offers a highly sensitive and reliable method for assessing gut permeability.
- This method enables reduced probe dosages, enhancing patient comfort and minimizing test-induced complications.
- The improved sensitivity and accuracy of this assay provide a valuable tool for clinical diagnostics and research in gastrointestinal function.