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Development and validation of a HPLC‐PDA bioanalytical method for the simultaneous estimation of Aliskiren and Amlodipine in human plasma
Author(s) -
Mannemala Sai Sandeep,
Nagarajan Janaki Sankarachari Krishnan
Publication year - 2015
Publication title -
biomedical chromatography
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.4
H-Index - 65
eISSN - 1099-0801
pISSN - 0269-3879
DOI - 10.1002/bmc.3279
Subject(s) - chemistry , chromatography , protein precipitation , amlodipine , hydrochlorothiazide , detection limit , high performance liquid chromatography , bioanalysis , extraction (chemistry) , human plasma , medicine , blood pressure , radiology
A simple, unique and selective HPLC‐PDA method was developed and validated for the simultaneous estimation of aliskiren (ALS) and amlodipine (AML) in human plasma. Extraction of the sample was accomplished by protein precipitation. Plasma proteins were precipitated by employing acetonitrile containing hydrochlorothiazide as internal standard. The compounds were analyzed by HPLC by using PDA detector on a Hibar C 18 (250 × 4.6 mm) column with a mobile phase comprising acetonitrile and phosphate buffer (pH 4.2 and 25 m m ; 60:40 v/v) with a flow rate of 0.8 mL/min. Different sample pretreatment techniques were evaluated but protein precipitation was found to be satisfactory, offering good recovery values of 97.11–98.45% for ALS and 97.5–99.12% for AML. The within‐day precisions for ALS were 96.66, 99.16 and 99.41% at 90, 240 and 480 ng/mL, respectively, and for AML they were 97.27, 99.54 and 99.31% at 3.3, 8.8 and 17.6 ng/mL, respectively. The between‐day precisions for ALS were 96.66, 99.16 and 99.41% at 90, 240 and 480 ng/mL, respectively and the between‐day precisions for AML were 98.18, 99.20 and 99.40% at 3.3, 8.8 and 17.6 ng/mL, respectively. The limit of quantitation was 30 and 1.0 ng/mL for ALS and AML respectively. Different constituents of plasma proteins did not interfere with the absolute recovery of ALS and AML. Copyright © 2014 John Wiley & Sons, Ltd.