Acta Pharm. 58 (2008) 243-256
Original research paper
Use of chemometrics for development and
validation of an RP-HPLC method for simultaneous determination of haloperidol
and related
compounds
RUMENKA PETKOVSKA and ANETA DIMITROVSKA
rupe@ff.ukim.edu.mk
Department of Chemistry,
Faculty of Pharmacy, “Ss. Cyril and Methodius” University, 1000 Skopje,
Macedonia
Accepted July 21, 2008
A rapid resolution reversed-phase high performance liquid chromatographic (RR RP-HPLC) method has been
developed and validated for simultaneous determination of haloperidol and six
related compounds. Investigated
matrix was a laboratory mixture of the
therapeutic active substance haloperidol and its six related compounds in a concentration
ratio 300:1. Experimental design was used during method optimization (full
factorial 23 design) and robustness testing (Central Composite
Circumscribed design). Three
factors: organic phase variation
during gradient elution, flow rate and gradient rise time were independent variables. To
estimate the system response during the optimization procedure and robustness
testing, resolution (Rs)
and a chromatographic response function (CRF)
were used. Chromatography was performed with the mobile phase containing
phosphate buffer pH 6.5 and acetonitrile as organic modifier. Separation was achieved using gradient
elution (organic phase fraction changed linearly from 20 to 72%) over 7 min. A Zorbax Eclipse XDB C18 Rapid
Resolution HT 4.6 mm × 50 mm, 1.8 μm particle size, column was used at 25 °C
at a flow rate of 1.5 mL min−1. UV detection was performed at
230 nm. The total time for chromatographic separation was 5.5 min with a total
analysis time of 7.0 min. The method was validated for its linearity, precision,
modal recovery and robustness.
Keywords: haloperidol, impurities, RP-HPLC, validation, experimental
design