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A simple and rapid ion-pair HPLC method for simultaneous quantitation of 4-nitrophenol and its glucuronide and sulfate conjugates.
J Biochem Biophys Methods. 2006 Nov 30; 69(1-2):43-50.JB

Abstract

Because of its simple and well characterized metabolic profile, 4-nitrophenol is widely used as a model substrate to investigate the influence of drug therapy, disease, nutrient deficiencies and other physiologically altered conditions on conjugative drug metabolism in animal studies. For simultaneous determination of 4-nitrophenol (PNP), 4-nitrophenyl-beta-D-glucuronide (PNP-G) and 4-nitrophenyl-sulfate (PNP-S) in samples generated in rat small intestine luminal perfusion experiments, an ion-pair HPLC assay coupled with UV detection was set up. The RP-HPLC separation was achieved with a methanol-water mixture (50:50, v/v) containing 0.01 M tetrabutyl-ammonium-bromide with UV detection of the analytes at 290 nm. The isocratic system was operated at ambient temperature and required less than 7 min of chromatographic time. The method provided good enough within-day precision, between-day precision and linearity in the target concentration ranges of 6-1200 microM (PNP) and 2.5-100 microM (PNP-G and PNP-S). The instrumental limit of quantification for PNP-G and PNP-S was found to be 2.7 microM and 2.1 microM, respectively. The assay was applied for determination of PNP, PNP-G and PNP-S in rat small intestine perfusates.

Authors+Show Affiliations

Institute of Pharmaceutical Chemistry, University of Pécs, Rókus str. 2, H-7624 Pécs, Hungary.No affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article

Language

eng

PubMed ID

16844228

Citation

Almási, Attila, et al. "A Simple and Rapid Ion-pair HPLC Method for Simultaneous Quantitation of 4-nitrophenol and Its Glucuronide and Sulfate Conjugates." Journal of Biochemical and Biophysical Methods, vol. 69, no. 1-2, 2006, pp. 43-50.
Almási A, Fischer E, Perjési P. A simple and rapid ion-pair HPLC method for simultaneous quantitation of 4-nitrophenol and its glucuronide and sulfate conjugates. J Biochem Biophys Methods. 2006;69(1-2):43-50.
Almási, A., Fischer, E., & Perjési, P. (2006). A simple and rapid ion-pair HPLC method for simultaneous quantitation of 4-nitrophenol and its glucuronide and sulfate conjugates. Journal of Biochemical and Biophysical Methods, 69(1-2), 43-50.
Almási A, Fischer E, Perjési P. A Simple and Rapid Ion-pair HPLC Method for Simultaneous Quantitation of 4-nitrophenol and Its Glucuronide and Sulfate Conjugates. J Biochem Biophys Methods. 2006 Nov 30;69(1-2):43-50. PubMed PMID: 16844228.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - A simple and rapid ion-pair HPLC method for simultaneous quantitation of 4-nitrophenol and its glucuronide and sulfate conjugates. AU - Almási,Attila, AU - Fischer,Emil, AU - Perjési,Pál, Y1 - 2006/05/07/ PY - 2005/10/31/received PY - 2006/03/18/revised PY - 2006/05/01/accepted PY - 2006/7/18/pubmed PY - 2006/12/9/medline PY - 2006/7/18/entrez SP - 43 EP - 50 JF - Journal of biochemical and biophysical methods JO - J Biochem Biophys Methods VL - 69 IS - 1-2 N2 - Because of its simple and well characterized metabolic profile, 4-nitrophenol is widely used as a model substrate to investigate the influence of drug therapy, disease, nutrient deficiencies and other physiologically altered conditions on conjugative drug metabolism in animal studies. For simultaneous determination of 4-nitrophenol (PNP), 4-nitrophenyl-beta-D-glucuronide (PNP-G) and 4-nitrophenyl-sulfate (PNP-S) in samples generated in rat small intestine luminal perfusion experiments, an ion-pair HPLC assay coupled with UV detection was set up. The RP-HPLC separation was achieved with a methanol-water mixture (50:50, v/v) containing 0.01 M tetrabutyl-ammonium-bromide with UV detection of the analytes at 290 nm. The isocratic system was operated at ambient temperature and required less than 7 min of chromatographic time. The method provided good enough within-day precision, between-day precision and linearity in the target concentration ranges of 6-1200 microM (PNP) and 2.5-100 microM (PNP-G and PNP-S). The instrumental limit of quantification for PNP-G and PNP-S was found to be 2.7 microM and 2.1 microM, respectively. The assay was applied for determination of PNP, PNP-G and PNP-S in rat small intestine perfusates. SN - 0165-022X UR - https://www.unboundmedicine.com/medline/citation/16844228/A_simple_and_rapid_ion_pair_HPLC_method_for_simultaneous_quantitation_of_4_nitrophenol_and_its_glucuronide_and_sulfate_conjugates_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0165-022X(06)00093-5 DB - PRIME DP - Unbound Medicine ER -