HPLC-Fluorescence Detection Method for Concurrent Estimation of Domperidone and Naproxen. Validation and Eco-Friendliness Appraisal Studies

Abstract

This work demonstrates a simple and reliable HPLC method with fluorimetric detection for simultaneous estimation of domperidone (DOM) and naproxen (NAP). Successful chromatographic separation was accomplished using Inertsil ODS C18 column (5 μm, 4.6 × 150 mm) with gradient elution of the mobile phase consisting of 0.01 M phosphate buffer (pH 5.5) solution and acetonitrile. The gradient elution started with 25% acetonitrile increased linearly to 65% in 5 min, then kept at this percentage till the end of the run. The mobile phase was pumped at a flow rate of 1.0 mL/min. The excitation wavelength at 284 nm was found suitable for both DOM and NAP since it corresponds to a maximum for the minor component DOM and measurable excitation for NAP, while using 316 and 355 nm as emission wavelengths for DOM and NAP, respectively. Peaks eluted with excellent resolution at retention times 4.4 and 6.3 min for DOM and NAP, respectively. Performance of the proposed method was tested according to ICH guidelines in regard to linearity, ranges, precision, accuracy, robustness, detection and quantitation limits. Calibration curves were linear in the ranges of 0.8-3.6 and 1.0-2.5 µg/mL for DOM and NAP respectively with correlation coefficients not less than 0.9996. The validated method was successfully applied to the analysis of DOM and NAP in their laboratory prepared tablets resembling the commercial dosage form, and assay results were favorably compared with a published reference HPLC method. The method's greenness was assessed using the Analytical Eco-Scale and the novel Analytical Greenness metric (AGREE).

Keywords: Domperidone; Fluorimetric detection; Greenness assessment; HPLC; Naproxen.

Fig. 1

Chemical structures of naproxen (NAP) and domperidone (DOM)

Fig. 2

HPLC chromatogram at λ_ex 284 nm and λ_em 316 nm of 20 μL injection volume of a mixture containing: (1) 2.0 μg/mL DOM and (2) 2.0 μg/mL NAP

Fig. 3

HPLC chromatograms at λex 284 nm of solution containing (a) 25 µg/mL NAP and 1 µg/mL DOM at λem 316 nm and (b) 2 µg/mL NAP and 0.08 µg/mL DOM at λem 355 nm obtained from laboratory prepared tablet solution