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Sci. Pharm. 2011, 79(4), 837-848; doi:10.3797/scipharm.1106-22

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.
Received: 26 June 2011 / Accepted: 18 August 2011 / Published: 18 August 2011
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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
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

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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.

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