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Article

HPLC Quantification of 4-Nitrophenol and its Conjugated Metabolites from Bile

1
Institute of Pharmaceutical Chemistry, University of Pécs, Rókus Street 2, H-7624 Pécs, Hungary
2
Department of Pharmacology, University of Pécs, Szigeti Street 12, H-7624 Pécs, Hungary
*
Author to whom correspondence should be addressed.
Sci. Pharm. 2011, 79(4), 837-848; https://doi.org/10.3797/scipharm.1106-22
Submission received: 26 June 2011 / Accepted: 18 August 2011 / Published: 18 August 2011

Abstract

An isocratic ion pair RP-HPLC method with UV-Vis detection has been developed and validated for simultaneous analysis of 4-nitrophenol (PNP), 4-nitrophenyl β-glucuronide (PNP-G), and 4-nitrophenyl sulfate (PNP-S) in rat bile samples using 4-ethylphenol (ETP) as internal standard. Chromatographic separation was achieved on a C18 column by isocratic elution with a mobile phase consisted of methanol-0.01 M citrate buffer pH 6.2 (47:53 v/v) containing 0.03 M TBAB. The flow rate was 1.0 ml min−1, the detection was affected at 290 nm. Calibration plots were generated over the concentration range 1–100 μM PNP, PNP-G, PNP-S with a common lower limit of quantification of 2.5 μM. Intra- and inter-day precision and repeatability were determined at six different concentrations. Results obtained by application of the method for determination of PNP, PNP-G and PNP-S in bile fractions collected during intestinal perfusion of PNP in hyperglycemic rats are presented.
Keywords: 4-Nitrophenol; Glucuronic acid conjugate; Sulfate conjugate; RP-HPLC; Intestinal metabolism 4-Nitrophenol; Glucuronic acid conjugate; Sulfate conjugate; RP-HPLC; Intestinal metabolism

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MDPI and ACS Style

ALMÁSI, A.; FISCHER, E.; PERJÉSI, P. HPLC Quantification of 4-Nitrophenol and its Conjugated Metabolites from Bile. Sci. Pharm. 2011, 79, 837-848. https://doi.org/10.3797/scipharm.1106-22

AMA Style

ALMÁSI A, FISCHER E, PERJÉSI P. HPLC Quantification of 4-Nitrophenol and its Conjugated Metabolites from Bile. Scientia Pharmaceutica. 2011; 79(4):837-848. https://doi.org/10.3797/scipharm.1106-22

Chicago/Turabian Style

ALMÁSI, Attila, Emil FISCHER, and Pál PERJÉSI. 2011. "HPLC Quantification of 4-Nitrophenol and its Conjugated Metabolites from Bile" Scientia Pharmaceutica 79, no. 4: 837-848. https://doi.org/10.3797/scipharm.1106-22

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