Simple and Reliable HPLC Method of Abacavir Determination in Pharmaceuticals, Human Serum and Drug Dissolution Studies from Tablets

Abstract

This work describes a new, fully validated, simple, rapid, selective, and sensitive HPLC method with UV detection for the direct determination of abacavir in pharmaceutical dosage forms, raw materials, spiked human serum, and drug dissolution studies without any time‐consuming extraction or evaporation steps prior to drug assay. The mobile phase employed was methanol∶acetonitrile: 0.015 M KH₂PO₄ (36∶2.6∶61.4 v/v/v) adjusted to pH 6.9 with 5 N NaOH. The samples of 20 µL were injected onto a Waters Spherisorb ODSI (250 × 4.6 mm, 5 µm particle size) column. Ketoprofen was used as internal standard. The flow rate was 1.0 mL min⁻¹. The retention times were 5.49 min for abacavir and 9.15 min for ketoprofen in mobile phase, 5.46 min for abacavir and 9.24 min for ketoprofen in serum samples. The samples were detected at 284 nm. The assay was linear in the concentration range 0.010–20 µgmL⁻¹ (r = 0.999) with a slope of 1.35 × 10⁻³; intercept of 0.0841 and the limit of detection was 0.00093 µg mL⁻¹ in mobile phase and 0.025–20 µg mL⁻¹ (r = 0.999) with a slope of 1.44 × 10⁻³; intercept of 0.0733 and the limit of detection was 0.00418 µg mL⁻¹ in human serum. The linearity of the detector response for abacavir was determined by plotting peak area ratios vs. concentration. It was successfully applied to the analysis of abacavir pharmaceutical preparations, and human serum samples without any interference by the excipients and endogenous substances. Moreover, the method can be used for the determination of abacavir for monitoring its concentration for in vitro dissolution studies.

Keywords:

• Abacavir determination
• drug dissolution study
• human serum
• pharmaceutical dosage form