article · Scientific Reports
New spectrofluorimetric techniques offer a sensitive, simple, and cost-effective way to measure pholcodine alone and alongside ephedrine. The first approach relies on measuring the native fluorescence of pholcodine at 337 nm after excitation at 284 nm, achieving low detection and quantitation limits across a range of 0.01 to 2.4 micrograms per millilitre. The second approach uses synchronous fluorimetry for the concurrent determination of pholcodine and ephedrine without mutual interference, recording emissions at 286 nm and 304 nm respectively. Both drugs showed strong linear responses at low concentrations. The synchronous method proved effective when tested on commercial syrup formulations. Furthermore, both methods functioned successfully in laboratory plasma samples, confirming their utility for bioanalytical testing and drug abuse screening. Both procedures met official international validation guidelines and demonstrated favourable environmental profiles according to standard greenness assessment tools.
Accurate and affordable measurement of active pharmaceutical ingredients is essential for quality control and toxicology. Pholcodine and ephedrine require careful monitoring, particularly in cases of potential drug abuse. By providing sensitive detection in biological fluids and pharmaceutical syrups using standard fluorescence equipment, these techniques present environmentally friendly alternatives to complex chromatographic testing.
These analytical procedures could be adopted by pharmaceutical quality control laboratories, clinical toxicology services, and drug abuse monitoring facilities. Tested on pharmaceutical syrup and laboratory-spiked plasma samples, the methods represent applied research ready for laboratory protocol integration. Because they rely on cost-effective spectrofluorimetry and follow standard international validation criteria, analytical testing providers could implement them directly without requiring expensive chromatographic machinery.
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In this study, sensitive, facile, and cost-effective spectrofluorimetric approaches were developed for the determination of pholcodine and ephedrine. Method I is a novel spectrofluorimetric method depending on measuring the native fluorescence of pholcodine at 337 nm after excitation at 284 nm over a concentration range of 0.01-2.4 μg/mL. The method sensitivity reached quantitation and detection limits down to 10.0 and 5.0 ng/mL, respectively. Method II relied on the simultaneous estimation of pholcodine and ephedrine using synchronous fluorimetry for the first time. The cited drugs were measured concurrently at 286 and 304 nm for pholcodine and ephedrine, respectively at Δλ of 40 nm without interference. Excellent linear relationship between concentration and response was obtained over the ranges of 0.05-6.0 μg/mL and 0.02-1.0 μg/mL for pholcodine and ephedrine, respectively. The method showed distinct sensitivity and exhibited quantitation limits of 20.0 and 10.0 ng/mL and detection limits of 10.0 and 5.0 ng/mL, respectively. The method was successfully applied to the syrup dosage form. The two developed approaches were also applied to in-vitro plasma samples, showing good bioanalytical applicability and providing further insights for monitoring drug abuse. The proposed methods were validated according to ICHQ2(R1) guidelines. The proposed methodologies' greenness profiles were evaluated using two greenness assessment tools.
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DOI: 10.1038/s41598-022-13194-1
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