Full Validation of an HPLC-UV Analytical Method for Azithromycin Quantification Using Comparative Approaches: Total...

Area of Science:

• Analytical Chemistry
• Pharmaceutical Analysis
• Method Validation

Background:

• Accurate quantification of Azithromycin is crucial for reliable dosage control in conventional release tablets.
• Analytical method validation ensures the precision and trustworthiness of quantification results for informed decision-making.
• Azithromycin, a complex molecule derived from erythromycin, necessitates rigorous analytical scrutiny.

Purpose of the Study:

• To validate an innovative method for Azithromycin quantification utilizing accuracy profiles.
• To compare measurement uncertainty calculations using two distinct formulas (Feinberg et al. and Saffaj & Ihssane) against the ISO GUM approach.
• To assess the robustness and accuracy of the proposed quantification method for pharmaceutical applications.

Main Methods:

• Liquid chromatography with a reversed-phase C18 stationary phase and UV detection at 210 nm.
• Isocratic elution using a mobile phase of acetonitrile and dipotassium hydrogen phosphate buffer (60:40, v/v) at pH 8 and 40°C.
• Application of accuracy profiles and calculation of measurement uncertainty using specified formulas and the ISO GUM approach.

Main Results:

• Accuracy profiles demonstrated that anticipated findings remained within the 95% tolerance interval and ±5% acceptance limits.
• Measurement uncertainty values derived from the total error approach formulas were comparable.
• The ISO GUM approach tended to overestimate expanded uncertainty compared to empirical estimation from validation data.

Conclusions:

• The total error approach facilitates accurate routine uncertainty closure, emphasizing comprehensive method validation.
• The proposed quantification method is robust for pharmaceutical use, exhibiting good accuracy within predefined limits.
• The method ensures 95% tolerance and uncertainty limits are within the acceptable ±5% range, supporting reliable Azithromycin dosage.