Development and Validation of a Stability-Indicating RP-HPLC Method for the Quantitative Analysis of Anagrelide Hydrochloride

Sudhakar S Pujeri

doi:10.3797/scipharm.1112-22

Abstract

A simple, rapid, and stability-indicating reverse-phase liquid chromatographic assay method was developed for Anagrelide Hydrochloride (ANG) in the presence of its degradation products generated from forced decomposition studies. The HPLC separation was achieved on a C18 Inertsil column (250 mm × 4.6 mm i.d. particle size is 5 μm), using solution A, a mixture of 0.03 M potassium di-hydrogen phosphate pH-adjusted to 3.0 using ortho-phosphoric acid (buffer): methanol: acetonitrile (90:5:5, v/v/v), and solution B, which contains a mixture of buffer: acetonitrile (10:90, v/v). The UV detector was operated at 251 nm while column temperature was maintained at 40°C, and the gradient program had the flow rate of 1.0 mL min−1. The developed method was validated as per ICH guidelines with respect to specificity, linearity, precision, accuracy, robustness, and limit of quantification. The method was found to be simple, specific, precise, accurate, and reproducible. Selectivity was validated by subjecting the stock solution of ANG to acidic, basic, photolysis, oxidative, and thermal degradation. The calibration curve was found to be linear in the concentration range of 0.05–152 μg mL−1 (R2 = 0.9991). The peaks of degradation products did not interfere with that of pure ANG. The utility of the developed method was examined by analyzing the tablets containing ANG.

Highlights

Anagrelide (6,7-dichloro-1,5-dihydroimidazo[2,1-b]quinazolin-2(3H)-one, fig. 1), is a potent blood platelet reducing agent
The ICH guidelines were followed to calculate the values of Limit of Detection (LOD) and Limit of Quantification (LOQ) and were observed to be 0.019 μg mL−1 and 0.054 μg mL−1, respectively, and obtained %RSD at LOQ level was 1.2
Significant degradation was observed in oxidation of ANG sample (Fig. 3), 15% major degradation product was observed at retention time of 4.069 min and having maximum absorbance at 195.9 and 262 nm

Summary

Introduction

Anagrelide (6,7-dichloro-1,5-dihydroimidazo[2,1-b]quinazolin-2(3H)-one, fig. 1), is a potent blood platelet reducing agent. Anagrelide (ANG) is a drug used for the treatment of essential thrombocytosis [1]. It works by inhibiting the maturation of megakaryocytes into platelets [2]. Anagrelide hydrochloride was approved by the FDA in 1997 for the treatment of patients with thrombocythemia, secondary to myeloproliferative disorders, to reduce the elevated platelet count and the risk of thrombosis and to ameliorate associated symptoms including thrombo-hemorrhagic events. Significant inhibition of platelet aggregation is observed only at doses of ANG higher than those required to reduce platelet count [3, 4]

Results

Discussion

Conclusion