Methods:
Sequential injection chromatography (SIC) with monolithic column has been
proposed with potential benefits for separation and quantification.
Objective:
To utilize SIC to develop a new assay method for the separation and quantification of some
phenothiazines (promethazine, chlorpromazine and perphenazine) in human urine and synthetic pharmaceutical
formulations.
Methods:
The 32 full-factorial design was adopted to study the effect of mobile phase composition on
separation efficiency, retention time, peak height and baseline. The separation was conducted on a C18
monolithic column (100 × 4.6 mm) using a mobile phase composition of phosphate:
acetonitrile:methanol (60:28:12) at pH 4.0. The detection was carried out using a miniaturized
fiber optic spectrometer at 250 nm.
Results:
Satisfactory analytical features, including number of theoretical plates (1809-6232), peak
symmetry (1.0-1.3), recovery (95.5-99.1% in pharmaceutical formulations and 91.6-94.7% in urine),
intra-day precision (0.36-1.60% for pharmaceutical formulation and 2.96-3.67 for urine), inter-day precision
(1.47-2.28% for pharmaceutical formulation), limits of detection (0.23-0.88 μg/ml) and limits of
quantification (0.77-2.90 μg/ml), were obtained.
Conclusion:
The remarkable advantages of the proposed SIC method are the inexpensiveness in terms
of instrumentation and reagent consumption.
