Optimization and validation of an HPLC method for the quantification of Etodolac in liposomes and assessment of its pharmacokinetic profile.

OBJECTIVE

Aim: To develop and evaluate high performance liquid chromatogram method to simultaneously quantify Etodolac in liposomes.

PATIENTS AND

METHODS

Materials and

Methods

The chromatographic separation was carried out on the C18 column, mobile phase of methanol: acetonitrile: water at a ratio of (30:60:10) and a flow rate of (1ml.min-1), with a detection wavelength of 225 nm. Liposomes were prepared by thin film hydrating method using Distearoylphosphatidylcholine and cholesterol.

RESULTS

Results

Linear concentration range of the assay was 2.5-25 µg.ml-1 with a regression coefficient factor R2 ≥ 0.999 with a retention time of Etodolac 4.11 ± 0.081 min. The method was rigorously validated for linearity, accuracy, precision, specificity, and stability, ensuring its reliability. The concentrations of Etodolac 2.5, 5, 7.5, and 10 µg.ml-1 were used, with the intraday n=6 and intraday n=6 precision and accuracy for Etodolac meeting the accepted criteria of the United States Food and Drug Administration guideline. The limited detection and limited quantification values for Etodolac were 0.42 µg.ml-1 and 1.3 µg.ml-1, respectively. Conventional liposomes have characterization (PZ 88 ± 0.02, PDI 0.012 ± 0.01, %EE 89.5 ± 0.004, and Z-potential -10.2mV ± 0.001), while PEGylated liposomes (PZ 92 ± 0.003, PDI 0.01 ± 0.02, %EE 94.3% ± 0.01, and Z-potential -13.6mV ± 0.012).

CONCLUSION

Conclusions

The developed and validated method of HPLC was successfully applied to determine Etodolac in bulk and liposomes for routine quantitative analysis.