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High-performance liquid chromatography method for the quantification of entacapone in human plasma
N V S Ramakrishna1, K N Vishwottam, S Wishu
1Biopharmaceutical Research, Suven Life Sciences Ltd., Serene Chambers, Road #7, Banjara Hills, Hyderabad 500034, India. nvsrk@suven.com
Summary
A new HPLC method accurately quantifies entacapone in human plasma. This validated method is suitable for pharmacokinetic studies of entacapone, an antiparkinsonian agent.
Area of Science:
- Analytical Chemistry
- Pharmacology
Background:
- Entacapone is a novel antiparkinsonian agent.
- Accurate quantification of entacapone in biological matrices is crucial for pharmacokinetic studies.
Purpose of the Study:
- To develop and validate a simple, sensitive, and selective High-Performance Liquid Chromatography (HPLC) method for entacapone quantification in human plasma.
Main Methods:
- Liquid-liquid extraction (LLE) using ethyl acetate/n-hexane.
- Reverse-phase C18 column chromatography with UV detection at 315 nm.
- Isocratic mobile phase: 30 mM phosphate buffer (pH 2.75)/acetonitrile (62/38, v/v).
Main Results:
- Lower limit of quantitation (LLOQ): 25 ng/mL with <8% relative standard deviation.
- Linear range: 25-2500 ng/mL.
- High precision (within-batch: 1.7-7.8%, between-batch: 2.2-4.2%) and accuracy (within-batch: 97.5-106.0%, between-batch: 98.7-107.5%).
- No interference from coadministered drugs and excellent analyte stability.
Conclusions:
- The developed HPLC method is validated, demonstrating high sensitivity, simplicity, and repeatability.
- This method is suitable for routine pharmacokinetic analyses of entacapone in human plasma.