Validation of an HPLC-DAD Method for Quercetin Quantification in Nanoparticles

Area of Science:

• Analytical Chemistry
• Pharmaceutical Sciences
• Nanotechnology

Background:

• Accurate quantification of quercetin is vital for pharmaceutical formulation development and quality control.
• Evaluating quercetin delivery systems and pharmacokinetics necessitates validated analytical methods.
• Existing methods may lack the sensitivity or efficiency required for complex matrices.

Purpose of the Study:

• To validate a high-performance liquid chromatography with diode array detection (HPLC-DAD) method for quercetin quantification in nanoparticles.
• To optimize mobile phase conditions and detection wavelengths for sensitive and selective quercetin analysis.
• To assess key validation parameters including precision, accuracy, linearity, sensitivity, stability, and selectivity.

Main Methods:

• Development and validation of an HPLC-DAD method.
• Testing various mobile phase compositions and detection wavelengths (254 nm and 368 nm).
• Assessment of method performance based on established validation parameters.

Main Results:

• Optimal analysis achieved at 368 nm with a mobile phase of 1.5% acetic acid and a water/acetonitrile/methanol ratio of 55:40:5.
• The method demonstrated excellent linearity (R² > 0.995), specificity, and repeatability (CV 2.4–6.7%).
• High accuracy (88.6–110.7%), low detection (0.046 µg/mL) and quantification (0.14 µg/mL) limits, and improved stability at 4 °C were observed.

Conclusions:

• The validated HPLC-DAD method is sensitive, efficient, and suitable for quantifying quercetin in nanoparticles.
• This method surpasses recent analytical approaches in bias assessment parameters and offers advantages over spectrophotometric techniques.
• The validated method provides a reliable tool for quality control in pharmaceutical development and future analysis in complex biological matrices.