Determination of p-aminohippuric acid in rat plasma by liquid chromatography-tandem mass spectrometry |
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Authors: | Hsien-Yuan Fan Chang-Ching LinLi-Heng Pao |
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Affiliation: | School of Pharmacy, National Defense Medical Center, Taipei, Taiwan, ROC |
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Abstract: | A rapid, simple and sensitive method was developed for the determination of para-aminohippuric acid (PAH) in rat plasma using liquid chromatography tandem mass spectrometry (LC-MS-MS). Acetaminophen was used as the internal standard. Chromatographic separation was performed using a Symmetry C18 column and the mobile phase was composed of A: 2 mM ammonium formate and 0.1% formic acid in water and B: 2 mM ammonium formate and 0.1% formic acid in acetonitrile (ACN) (A:B, 30:70, v/v). Detection was performed on a triple–quadrupole tandem mass spectrometer using positive ion mode electrospray ionization (ESI) in the multiple reaction monitoring (MRM) mode. The MS/MS ion transitions monitored were m/z 195.2 → 120.2 and 152.1 → 110.1 for PAH and acetaminophen, respectively. Good linearity is observed over the concentration range of 0.1–500 μg/ml. The method was proved to be accurate and reliable and was applied to a pharmacokinetic study in rat. |
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Keywords: | Para-aminohippuric acid Liquid chromatography-tandem mass spectrometry Pharmacokinetics Analytical validation Acetaminophen Rat plasma |
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