Determination of Acetaminophen in Pharmaceutical Preparations and Body Fluids by High-Performance Liquid Chromatography with Electrochemical Detection

Riggin, R.M.; Schmidt, Allen; Kissinger, Peter T.

doi:10.1002/jps.2600640423

Cited by 79 publications

(20 citation statements)

References 23 publications

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“…Another advantage is that the assay uses tyrosine as an internal standard, eliminating the need to determine the absolute amount of protein hydrolyzed. This assay also can detect and quantify APAP, as others have previously reported (Riggin et al, 1975;Munson et al, 1978;Miner and Kissinger, 1979). However, dialysis or protein precipitation before proteolysis is effective in removing any contamination of free compounds.…”

Section: Discussionmentioning

confidence: 89%

“…Acetaminophen-and glutathione-derived metabolites have previously been assayed by HPLC-ECD methods (Riggin et al, 1975;Munson et al, 1978;Miner and Kissinger, 1979;Hall et al, 1986;Whitcomb and Block, 1994;Alonso et al, 1995;Bonkovsky, 1995;Rivera-Penera et al, 1997). The advantage of the HPLC-ECD assay for acetaminophen-cysteine in proteins is its ability to quantify conjugates in serum protease digestion.…”

mentioning

confidence: 99%

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Determination of Acetaminophen-Protein Adducts in Mouse Liver and Serum and Human Serum after Hepatotoxic Doses of Acetaminophen Using High-Performance Liquid Chromatography with Electrochemical Detection

Muldrew¹,

James²,

Coop³

et al. 2002

Drug Metab Dispos

178

174

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This article is available online at http://dmd.aspetjournals.org ABSTRACT:Acetaminophen-induced hepatotoxicity has been attributed to covalent binding of the reactive metabolite N-acetyl-p-benzoquinone imine to cysteine groups on proteins as an acetaminophen-cysteine conjugate. We report a high-performance liquid chromatography with electrochemical detection (HPLC-ECD) assay for the conjugate with increased sensitivity compared with previous methods. Previous methods to quantitate the protein-bound conjugate have used a competitive immunoassay or radiolabeled acetaminophen. With HPLC-ECD, the protein samples are dialyzed and then digested with protease. The acetaminophen-cysteine conjugate is then quantified by HPLC-ECD using tyrosine as an internal reference. The lower limit of detection of the assay is approximately 3 pmol/mg of protein. Acetaminophen protein adducts were detected in liver and serum as early as 15 min after hepatotoxic dosing of acetaminophen to mice. Adducts were also detected in the serum of acetaminophen overdose patients. Analysis of human serum samples for the acetaminophen-cysteine conjugate revealed a positive correlation between acetaminophen-cysteine conjugate concentration and serum aspartate aminotransferase (AST) activity or time. Adducts were detected in the serum of patients even with relatively mild liver injury, as measured by AST and alanine aminotransferase. This assay may be useful in the diagnostic evaluation of patients with hepatotoxicity of an indeterminate etiology for which acetaminophen toxicity is suspect.Acetaminophen (N-acetyl-p-aminophenol; APAP 1 ; Paracetamol) is the most commonly used drug for the treatment of pain and fever. Although safe at therapeutic doses, in overdose, acetaminophen produces severe hepatotoxicity. The mechanism of toxicity is by initial metabolism to the reactive metabolite N-acetyl-p-benzoquinone imine (NAPQI) by cytochrome P450 . At therapeutic doses, the reactive metabolite is detoxified by glutathione, but following overdoses, glutathione is depleted and the metabolite covalently binds to proteins as 3-(cystein-S-yl)-acetaminophen (APAP-CYS) (Streeter et al., 1984). Covalent binding to protein is thought to be a critical step in the development of hepatotoxicity.A number of methods have been used to quantify the amount of acetaminophen covalently bound to proteins. In initial studies showing the correlation between acetaminophen toxicity and covalent binding, Jollow and coworkers (1973) used radiolabeled acetaminophen. Subsequently, polyclonal antisera were raised that recognized the acetaminophen-cysteine adducts (Bartolone et al., 1987;Roberts et al., 1987), and the relationship between covalent binding and toxicity was studied extensively. coworkers (1989, 1990) developed a competitive enzyme-linked immunosorbent assay to quantify acetaminophen covalent binding in the liver and serum of treated mice. In addition, Western blot assays were performed. Although these latter assays were not quantitative, they were useful in comparing t...

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Section: Discussionmentioning

confidence: 89%

mentioning

confidence: 99%

Determination of Acetaminophen-Protein Adducts in Mouse Liver and Serum and Human Serum after Hepatotoxic Doses of Acetaminophen Using High-Performance Liquid Chromatography with Electrochemical Detection

Muldrew¹,

James²,

Coop³

et al. 2002

Drug Metab Dispos

178

174

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show abstract

“…Methods reporting the sensitivity necessary to quantify pharmacokinetics acetaminophen levels accurately have also been published [280][281][282]. Most of these methods used HPLC in conjunction with electrochemical detection, solid-phase extraction and sample extraction procedures that involve multiple time-consuming organic extractions and solvent evaporation steps [282][283][284][285]. Only a few of these methods [280,282,286] are available for rapid measuring the large number of samples that have to be analysed in a pharmacokinetic study.…”

Section: Biological Fluidsmentioning

confidence: 99%

Determination of paracetamol: Historical evolution

Bosch

Ruiz-Sánchez

Rojas

et al. 2006

Journal of Pharmaceutical and Biomedical Analysis

215

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“…This system involved anionexchange chromatography with detection and quantitation of paracetamol and its metabolites using both ultraviolet and cerate oxidimetric detectors, the cycle time for the process being 24 h per sample. Riggin et al (1975) utilised a rather unusual detector system, a thin layer electrochemical transducer. Plasma proteins were precipitated with perchloric acid and the supernatant was extracted with ethyl acetate.…”

Section: High-performance Liquid Chromatographymentioning

confidence: 99%

A Review of Methods for Plasma Paracetamol Estimation

Wiener

1978

Ann Clin Biochem

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Determination of Acetaminophen in Pharmaceutical Preparations and Body Fluids by High-Performance Liquid Chromatography with Electrochemical Detection

Cited by 79 publications

References 23 publications

Determination of Acetaminophen-Protein Adducts in Mouse Liver and Serum and Human Serum after Hepatotoxic Doses of Acetaminophen Using High-Performance Liquid Chromatography with Electrochemical Detection

Determination of Acetaminophen-Protein Adducts in Mouse Liver and Serum and Human Serum after Hepatotoxic Doses of Acetaminophen Using High-Performance Liquid Chromatography with Electrochemical Detection

Determination of paracetamol: Historical evolution

A Review of Methods for Plasma Paracetamol Estimation

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