SEQUENTIAL INJECTION CHROMATOGRAPHY FOR BIOFLUIDIC ANALYSIS: APPLICATION TO PROMETHAZINE ASSAY

Abstract

In the current study, a newly developed sequential injection chromatograph (SIC) system is introduced to biochemical analysis. A new SIC method for promethazine assay is provided. The method was optimized, validated, and applied to human urine and serum, in addition to pharmaceutical formulations. Trifluphenazine was used as an internal standard. The 2³ full-factorial design was adopted to screen the effect of mobile phase composition on separation efficiency, retention time, peak height, and baseline. The separation was conducted onto C18 monolithic column (4.6 × 25 mm) using 30 mmol/L phosphate:acetonitrile (50:50, v/v) at pH 4.0. The detection was carried out by miniaturized fiber optic spectrometric devices set at 250 nm. The limit of detection in biofluids was ≤23 ng/mL. The sensitivity was improved by optimizing the mobile phase composition, using a short monolithic column and a large sample volume (30 μL), in addition to preconcentration obtained by solid-phase and liquid–liquid extractions. Additionally, good recoveries (90.7–98.3%) in all types of samples were obtained. The remarkable advantages of the proposed SIC method are rapidity and simplicity, in addition to affordability in terms of instrumentation cost and reagent consumption.

Keywords:

• biofluidic analysis
• chemometric optimization
• method validation
• pharmaceutical analysis
• promethazine
• sequential injection chromatography

ACKNOWLEDGMENT

The author expresses his gratitude to the financial support from King Abdulaziz City for Science and Technology, Saudi Arabia, award # MT-3-6. The author also thanks the Department of Chemistry, College of Science, King Khalid University for allowing him to conduct a portion of this research.

Notes

Note: −1 and +1 refer to the minimum and maximum, respectively, as identified in Table 2.

^a Concentration.

^b Absorbance unit.

^a Recovery was calculated using spiked concentration as a reference value.

^b Relative standard deviation for four replicates.

^c Recovery was calculated using reference value obtained from the results of the British Pharmacopeia as a reference value.