Sequential Injection Chromatography for Separation and Quantification of Chlorpromazine in Human Urine and Pharmaceutical Formulations

Abstract

Sequential injection chromatography (SIC) is a recent, simple, and inexpensive green miniaturized separation technique. In the current study, SIC was exploited for the first time for biochemical analysis. A new SIC method for the separation and quantification of chlorpromazine in human urine, as well as pharmaceutical formulations, was developed and validated. Clozapine was used as an internal standard. Chlorpromazine was successfully separated on a C18 monolithic column (25x4.6 mm id). The UV detection was carried out at 250 nm using miniaturized fiber optic spectrometric devices. The optimum mobile phase composition was 30 mmol/L phosphate-acetonitrile-methanol (55.0 + 31.5 + 13.5, v/v/v) at pH 3.0. The sample volume was 40 microL and flow rate was 40 microL/s. Acceptable chromatographic results were obtained. The resolution was 2.7, peak symmetry was 1.1, and number of theoretical plates was more than 1 x 10(6). Good linearity (r = 0.9997) in the range of 25-100 microg/mL was also obtained. The method offered acceptable recovery for both human urine (89.6-93.1%) and pharmaceutical formulations (96.9-98.5%), which was sensitive enough to detect chlorpromazine. The LOD and LOQ in human urine were 61 and 204 ng/mL, respectively. The method was rapid and reagent-saving, and hence safe for the environment. The sample throughput was 26.3 samples/h and the total volume of consumed reagents was 4.0 mL.