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Microdosing Assessment to Evaluate Pharmacokinetics and Drug Metabolism in Rats Using Liquid Chromatography-Tandem Mass Spectrometry

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Abstract

Purpose

To evaluate the sensitivity requirement for LC-MS/MS as an analytical tool to support human microdosing study with sub-pharmacological dose, investigate proportionality of pharmacokinetics from the microdose to therapeutic human equivalent doses in rats and characterize circulating metabolites in rats administered with the microdose.

Materials and Methods

Five drugs of antipyrine, metoprolol, carbamazepine, digoxin and atenolol were administered orally to male Sprague–Dawley rats at 0.167, 1.67, 16.7, 167 and 1,670 μg/kg doses. Plasma samples were extracted using either solid phase extraction or liquid–liquid extraction, and analyzed using LC-MS/MS.

Results

Using 100 μl of plasma sample, the lower limit of quantitation for antipyrine (10 pg/ml), carbamazepine (1 pg/ml), metoprolol (5 pg/ml), atenolol (20 pg/ml), and digoxin (5 pg/ml) were achieved using an API 5000™. Proportional pharmacokinetics were observed from 0.167 μg/kg to 1,670 μg/kg for antipyrine and carbamazepine and from 1.67 to 1,670 μg/kg for atenolol and digoxin, while metoprolol exhibited a non-proportional pharmacokinetics relationship. Several metabolites of carbamazepine were characterized in plasma from rats dosed at 1.67 μg/kg using LC-MS/MS.

Conclusions

This study has shown the promise of sensitive LC-MS/MS method to support microdose pharmacokinetics and drug metabolism studies in human.

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Acknowledgments

The authors wish to thank Mr. Van Dinh and his group at Allergan for providing animal dosing and blood sample collection.

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Authors and Affiliations

  1. Department of Drug Safety Evaluation, Allergan, 2525 Dupont Drive, Irvine, California, 92612, USA

    Jinsong Ni, Hui Ouyang, Lisa Borbridge, Devin Welty & Andrew Acheampong

  2. Product Application Lab, Applied Biosystem/MDS Sciex, Concord, Ontario, Canada

    Mauro Aiello, Carmai Seto, Takeo Sakuma & Robert Ellis

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  1. Jinsong Ni
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  2. Hui Ouyang
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  3. Mauro Aiello
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  4. Carmai Seto
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  5. Lisa Borbridge
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  6. Takeo Sakuma
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  7. Robert Ellis
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  8. Devin Welty
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  9. Andrew Acheampong
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Correspondence to Jinsong Ni.

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Ni, J., Ouyang, H., Aiello, M. et al. Microdosing Assessment to Evaluate Pharmacokinetics and Drug Metabolism in Rats Using Liquid Chromatography-Tandem Mass Spectrometry. Pharm Res 25, 1572–1582 (2008). https://doi.org/10.1007/s11095-008-9555-x

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  • DOI: https://doi.org/10.1007/s11095-008-9555-x

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