Modified colorimetric method of plasma prednisolone

Area of Science:

• Analytical Chemistry
• Pharmacology
• Veterinary Medicine

Background:

• Prednisolone is a widely used corticosteroid in veterinary medicine.
• Accurate quantification of prednisolone in biological matrices is crucial for therapeutic monitoring and pharmacokinetic studies.
• Existing analytical methods may lack the simplicity, speed, or sensitivity required for routine analysis.

Purpose of the Study:

• To develop and validate a simple, precise, rapid, and sensitive colorimetric method for the quantitative analysis of prednisolone in dog plasma.
• To adapt and optimize a paper chromatographic procedure using thin-layer chromatography (TLC) for this purpose.

Main Methods:

• Dog plasma samples were extracted using methylene chloride after separation from cellular elements.
• Extracted prednisolone was purified through acid-base washing and concentrated.
• The concentrated sample was applied to a thin-layer chromatography plate coated with microcrystalline cellulose.
• Bands were visualized under UV light, scraped, and eluted with methanol.
• The eluate was reacted with the Porter-Silber reagent (phenylhydrazine) for colorimetric detection.
• Absorbance was measured at 410 nm.

Main Results:

• The adapted method demonstrated high recovery rates, with a mean of 97.5%.
• The method exhibited good precision, indicated by a coefficient of variation of 5.13%.
• The colorimetric analysis at 410 nm provided a sensitive and specific measure of prednisolone concentration.

Conclusions:

• The developed colorimetric method is suitable for the accurate and reliable quantitative analysis of prednisolone in dog plasma.
• This method offers a practical alternative for therapeutic drug monitoring and pharmacokinetic research in veterinary settings.
• The technique combines the advantages of chromatography with a sensitive colorimetric detection system.