Development and validation of a stability-indicating RP-HPLC method for the detection and quantification of azithromycin in bulk, and self-emulsifying drug delivery system (SEDDs) formulation

Abstract

The objective of this work is to develop and validate a simple, rapid and specific reverse phase HPLC-UV method for the determination of azithromycin (AZM) in bulk, and self-emulsifying drug delivery system (SEDDs). The separation was done using Hypersil GOLD C-18 analytical column packed with deactivated silica (250 mm x 4.6 mm ID x 5 µm) kept at 60 °C, ammonium acetate solution (30 mmolL −1 , pH= 6.8) and acetonitrile (18:82, v/v) as the mobile phase, and UV detection at 210 nm. Samples were eluted isocratically at a flow rate of 0.7 mL min −1 .Forced degradation studies on AZM in bulk and the developed formulation were carried out. The method was validated for system suitability, specificity, linearity, precision, and accuracy. Theoretical plates (N > 1500), tailing factor (T ≤ 1.5), and resolution (Rs > 3) were as per United States Pharmacopeia (USP). There were no interferences by SEDDs excipients and AZM degradation products. The linearity was observed over the concentration range of 5-200 μg mL -1 (R 2 > 0.9999).The limit of detection (LOD) and limit of quantification (LOQ) were 0.476 µg mL -1 and 1.443 µgmL -1 respectively. the developed method was statistically confirmed to be accurate, precise, and reproducible.