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  • Simultaneous gas-chromatographic urinary measurement of sugar probes to assess intestinal permeability: use of time course analysis to optimize its use to assess regional gut permeability.

Simultaneous gas-chromatographic urinary measurement of sugar probes to assess intestinal permeability: use of time course analysis to optimize its use to assess regional gut permeability.

Clinica chimica acta; international journal of clinical chemistry (2015-01-17)
Maliha Shaikh, Kumar Rajan, Christopher B Forsyth, Robin M Voigt, Ali Keshavarzian
ABSTRACT

Measurement of intestinal permeability is important in several diseases but currently several methods are employed. We sought to: (1) develop a new GC based method to measure urinary mannitol, lactulose and sucralose to assess regional and total gut permeability; (2) analyze the kinetics of these sugars in the urine to determine which ratio is useful to represent intestinal permeability; and (3) determine whether age, gender, race and BMI impact these values. Subjects drank a cocktail of sucrose, lactulose, mannitol and sucralose and these sugars were measured in the urine at 5, 12 and 24h with gas chromatography. Urinary mannitol exhibited significantly different kinetics than lactulose and sucralose which were similar to each other and varied little over the 24h. No permeability differences were observed for renal function, age, race, sex, or BMI. Our data do not support the use of the widely used L/M ratio as an accurate estimate of intestinal permeability. Our data support the use of: the sucralose/lactulose (S/M) ratio to measure: small intestine permeability (first 5h); small and large intestine (first 12h), and total gut permeability (24h). This was also found to be true in a Parkinson's disease model.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Acetic anhydride
Lactulose, European Pharmacopoeia (EP) Reference Standard
Sucrose, European Pharmacopoeia (EP) Reference Standard
Supelco
Lactulose, Pharmaceutical Secondary Standard; Certified Reference Material
USP
Lactulose, United States Pharmacopeia (USP) Reference Standard
USP
Glacial acetic acid, United States Pharmacopeia (USP) Reference Standard
Millipore
Bifido Selective Supplement B, suitable for microbiology
Supelco
5α-Androstan-17β-ol-3-one, VETRANAL®, analytical standard
Sigma-Aldrich
Acetic anhydride, Arxada quality, ≥99.5% (GC)
Sigma-Aldrich
Acetic anhydride, ACS reagent, ≥98.0%
USP
Sucrose, United States Pharmacopeia (USP) Reference Standard
Sigma-Aldrich
Acetic anhydride, 99.5%
Sigma-Aldrich
Acetic anhydride, ReagentPlus®, ≥99%
Supelco
Acetic anhydride, derivatization grade (GC derivatization), LiChropur, ≥99.0%
Millipore
Sucrose, suitable for microbiology, ACS reagent, ≥99.0%
Sigma-Aldrich
Sucrose, puriss., meets analytical specification of Ph. Eur., BP, NF
Sigma-Aldrich
Lactulose, ≥98.0% (HPLC)
Sigma-Aldrich
5α-Androstan-17β-ol-3-one, purum, ≥99.0% (TLC)
Sigma-Aldrich
Sucrose, BioUltra, Molecular Biology, ≥99.5% (HPLC)
Sigma-Aldrich
5α-Androstan-17β-ol-3-one, ≥97.5%
Sigma-Aldrich
Sucrose, ≥99.5% (GC), BioReagent, suitable for cell culture, suitable for insect cell culture
Sigma-Aldrich
Sucrose, ACS reagent
Supelco
Sucrose, analytical standard, for enzymatic assay kit SCA20
Sigma-Aldrich
Sucrose, ≥99.5% (GC), BioXtra
Sigma-Aldrich
Sucrose, ≥99.5% (GC), Grade II, suitable for plant cell culture
Sigma-Aldrich
Sucrose, ≥99.5% (GC)
Sigma-Aldrich
Sucrose, ≥99.5% (GC)
Sigma-Aldrich
Sucrose, Molecular Biology, ≥99.5% (GC)
Sigma-Aldrich
Sucrose, ≥99% (GC), Grade I, suitable for plant cell culture
Sigma-Aldrich
Lactulose, ≥95% (HPLC)