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Journal of Pharmaceutical Research International, ISSN: 2456-9119, ISSN: 2231-2919 (past),Vol.: 22, Issue.: 6


A Novel HPLC Method for Determination of Phenytoin in Human Plasma


Jesse Flores1, Sheril Alexander1 and Mariana Babayeva1*

1Department of Biomedical and Pharmaceutical Sciences, Touro College of Pharmacy, New York, USA.

Article Information


(1) Dr. Triveni Krishnan, National Institute of Cholera and Enteric Diseases, India.


(1) Minaketan Sahoo, Institute of Pharmacy & Technology, Salipur, India.

(2) Sheikh Shahnawaz, India.

(3) Márcio Rodrigues, Polytechnic of Guarda, Portugal.

(4) Alexander Chernonosov, Institute of Chemical Biology and Fundamental Medicine SB RAS, Russia.

(5) Abhishek Raj, India.

Complete Peer review History: http://www.sciencedomain.org/review-history/25422


Aim: Aim of this research was to develop and validate a simple, efficient and reproducible high-performance liquid chromatography method to measure phenytoin concentrations in human plasma

Study Design: Linearity, selectivity, sensitivity, accuracy and precision of the analytical methods were validated according to ICH guidelines.

Methodology: The method employed a Phenomenex C18 column kept at 25ºC. The mobile phase consisted of a 0.05 M potassium dihydrogen phosphate buffer solution (pH 2.8) and methanol in a ratio of 60:40, respectively. The flow rate of the mobile phase was 0.7 mL/min. Phenytoin was detected at a wavelength of 250 nm.

Results: Phenytoin was eluted at 7.4 minutes with no interference with the other components of the human plasma. The method was linear in the range of 1-25 μg/mL (R2 = 0.9998). The LOD and LOQ were calculated as 0.07 μg/mL and 0.20μg/mL, respectively. Recovery, tested in the range of 5-20 μg/mL, was found to be 99.21 - 102.09%. Intraday and interday precision RSDs were 0.65 and 0.29%, respectively.

Conclusion: The proposed method is fast, reliable and reproducible, and can be recommended to measure free phenytoin levels in the blood.

Keywords :

Phenytoin; HPLC; validation; human plasma.

Full Article - PDF    Page 1-7

DOI : 10.9734/JPRI/2018/41885

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