A Simple, Robust, and Convenient HPLC Assay for Urinary Lactulose and Mannitol in the Dual Sugar Absorption Test

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dc.citation.issue9
dc.citation.volume27
dc.contributor.authorSequeira IR
dc.contributor.authorKruger MC
dc.contributor.authorHurst RD
dc.contributor.authorLentle RG
dc.contributor.editorLocatelli M
dc.coverage.spatialSwitzerland
dc.date.accessioned2023-07-18T03:22:18Z
dc.date.accessioned2023-09-04T01:40:45Z
dc.date.available2022-04-21
dc.date.available2023-07-18T03:22:18Z
dc.date.available2023-09-04T01:40:45Z
dc.date.issued2022-05
dc.date.updated2023-07-13T00:23:53Z
dc.descriptionCopyright: © 2022 by the authorsen_US
dc.description.abstractBACKGROUND: Heterogeneous laborious analytical methodologies for the determination of urinary lactulose and mannitol limit their utility in intestinal permeability testing. METHODS: We developed an assay using a Shimadzu HPLC system, an Aminex HPX87C column, and refractive index detection. The test was calibrated using a series of dilutions from standard stock solutions of lactulose and mannitol 'spiked' into urine samples. The utility to quantify urinary excretion during the dual sugar absorption test over 6 h was also determined. RESULTS: Lactulose and mannitol were eluted isocratically at 5.7 and 10.1 min, respectively, with water as a mobile phase at a flow rate of 0.3 mL min-1, 858 psi, 60 °C. The calibration curves for both sugars were linear up to 500 µg mL-1 with a limit of detection in standard solutions at 4 µg mL-1 and in 'spiked' urine samples at 15 µg mL-1. The intra-assay and inter-assay CVs were between 2.0-5.1% and 2.0-5.1% for lactulose and 2.5-4.4% and 2.8-3.9% for mannitol. The urinary profiles of the 6 h absorption of lactulose and mannitol showed similar peak-retention times to standard solutions and were well-resolved at 5.9 and 10.4 min, respectively. CONCLUSIONS: The assay was easy to automate, using commonly available equipment and convenient requiring no prior laborious sample derivatization. The simplicity, reproducibility, and robustness of this assay facilitates its use in routine clinical settings for the quantification of intestinal permeability.
dc.description.confidentialfalse
dc.edition.editionMay 2022
dc.format.extent2677-
dc.identifier2677
dc.identifiermolecules27092677
dc.identifierhttps://www.ncbi.nlm.nih.gov/pubmed/35566024
dc.identifier.citationSequeira IR, Kruger MC, Hurst RD, Lentle RG. (2022). A Simple, Robust, and Convenient HPLC Assay for Urinary Lactulose and Mannitol in the Dual Sugar Absorption Test.. Molecules. 27. 9. (pp. 2677-).
dc.identifier.doi10.3390/molecules27092677
dc.identifier.eissn1420-3049
dc.identifier.elements-typejournal-article
dc.identifier.harvestedMassey_Dark
dc.identifier.issn1420-3049
dc.identifier.urihttp://hdl.handle.net/10179/19873
dc.languageeng
dc.publisherMDPI (Basel, Switzerland)
dc.publisher.urihttps://www.mdpi.com/1420-3049/27/9/2677
dc.relation.isPartOfMolecules
dc.rightsCC BYen_US
dc.rights.urihttps://creativecommons.org/licenses/by/4.0/en_US
dc.subjectdual sugar absorption test
dc.subjecthigh-performance liquid chromatography
dc.subjectintestinal permeability
dc.subjectlactulose
dc.subjectmannitol
dc.subjectChromatography, High Pressure Liquid
dc.subjectIntestinal Absorption
dc.subjectLactulose
dc.subjectMannitol
dc.subjectPermeability
dc.subjectReproducibility of Results
dc.titleA Simple, Robust, and Convenient HPLC Assay for Urinary Lactulose and Mannitol in the Dual Sugar Absorption Test
dc.typeJournal article
pubs.elements-id453159
pubs.organisational-groupOther
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