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Sci. Pharm. 2012, 80(1), 89-100; doi:10.3797/scipharm.1109-03

Validated HPLC Method for Concurrent Determination of Antipyrine, Carbamazepine, Furosemide and Phenytoin and its Application in Assessment of Drug Permeability through Caco-2 Cell Monolayers

1
Department of Pharmaceutical Technology (Formulations), National Institute of Pharmaceutical Education and Research (NIPER), S.A.S. Nagar, Mohali, Punjab-160 062, India
2
Department of Pharmaceutics, National Institute of Pharmaceutical Education and Research (NIPER), S.A.S. Nagar, Mohali, Punjab-160 062, India
*
Author to whom correspondence should be addressed.
Received: 16 September 2011 / Accepted: 22 October 2011 / Published: 22 October 2011
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Abstract

The present work explains the development and validation of a simple, rapid and sensitive liquid chromatographic method for the simultaneous determination of antipyrine (ANT), carbamazepine (CBZ), furosemide (FSD) and phenytoin (PHTN). Chromatographic analysis was carried out by a reversed phase technique on a C18 column, using water pH 3.0 and 50:50 mixtures of methanol and acetonitrile (58:42 v/v) as the mobile phase, at a flow-rate of 1.0 ml/min and a column temperature of 40°C. Detection was carried out at 205 nm for CBZ and PHTN and at 230 nm for ANT and FSD. The proposed method was evalu-ated for validation parameters including linearity, range, accuracy, precision, limit of detection (LOD), limit of quantification (LOQ) and specificity. Elution of drugs ANT, FSD, PHTN, and CBZ was observed at 4.1, 5.1, 12.3 and 13.5 min, respectively. The method was found to be linear (R2 ≥ 0.999) in the concen-tration range of 5–100 μM, with an acceptable accuracy and relative standard deviation. Results of intra- and inter-day validation (n=3) showed the method to be efficient for routine determination of these permeability markers in Caco-2 cell monolayer permeability studies. The method was successfully utilized for determination of standard compounds in Caco-2 permeability experiments.
Keywords: Validation; HPLC; Caco-2 cell monolayer; Phenazone; Carbamazepine; Furosemide; Phenytoin Validation; HPLC; Caco-2 cell monolayer; Phenazone; Carbamazepine; Furosemide; Phenytoin
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

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MDPI and ACS Style

PATIL, S.R.; KUMAR, L.; KOHLI, G.; BANSAL, A.K. Validated HPLC Method for Concurrent Determination of Antipyrine, Carbamazepine, Furosemide and Phenytoin and its Application in Assessment of Drug Permeability through Caco-2 Cell Monolayers. Sci. Pharm. 2012, 80, 89-100.

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