Transl Clin Pharmacol.  2016 Mar;24(1):30-36. 10.12793/tcp.2016.24.1.30.

Development and validation of a UPLC-MS/MS method for the quantification of acetaminophen in human plasma and its application to pharmacokinetic studies

• Cha J
• Kim BK
• Gwon MR
• Lee J
• Ohk B
• Kang WY
• Lim MS
• Seong SJ
• Kim HJ
• Lee HW
• Yoon YR

• KMID: 2383597
• DOI: http://doi.org/10.12793/tcp.2016.24.1.30

Abstract

We developed an ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method for the determination of acetaminophen concentration in human plasma. Following protein precipitated extraction, the analytes were separated and analyzed using an UPLC-MS/MS in the multiple reaction monitoring (MRM) mode with the respective [M+H]+ ions, m/z 152.06 → 110.16 for acetaminophen and m/z 180.18 → 138.12 for phenacetin (internal standard, IS). The method showed a linear response from 1 to 100 µg/mL (r > 0.9982). The limit of quantitation for acetaminophen in plasma was 1 µg/mL. The intra- and inter-day accuracy ranged in the ranges of 94.40-99.56% and 90.00-99.20%, respectively. The intra- and inter-day precision ranged in the ranges of 2.64-10.76% and 6.84-15.83%, respectively. This method was simple, reliable, precise and accurate and can be used to determine the concentration of acetaminophen in human plasma. Finally, this fully validated method was successfully applied to a pharmacokinetic study of acetaminophen in healthy volunteers following oral administration.

Keyword

Acetaminophen; Human plasma; UPLC-MS/MS; Validation; Pharmacokinetic study

MeSH Terms

Acetaminophen*
Administration, Oral
Healthy Volunteers
Humans*
Ions
Mass Spectrometry
Phenacetin
Plasma*
Acetaminophen
Ions
Phenacetin

Figure

• 

  Figure 1. Product ion mass spectra are given for (a) acetaminophen and (b) phenacetin (IS).

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  Figure 2. Typical MRM chromatograms of acetaminophen (upper panel) and phenacetin (lower panel) in human plasma samples obtained from (a) a drug-free blank plasma sample, (b) a plasma sample spiked with acetaminophen at the LLOQ (1 μg/mL) and 1 μg/mL phenacetin and (c) plasma from a volunteer 1 h after oral administration of a 1,000 mg dose of acetaminophen spiked with phenacetin (experimental conditions were same as in text).

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  Figure 3. Calibration curve of acetaminophen.

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  Figure 4. Mean plasma concentration-time profile of acetaminophen after a 1,000 mg oral dose in 8 healthy volunteers. The vertical bars show the standard errors.

Reference

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