A rapid stability-indicating, fused-core HPLC method for simultaneous determination of beta-artemether and lumefantrine in anti-malarial fixed dose combination products

作者：Suleman Sultan; Vandercruyssen Kirsten; Wynendaele Evelien; D' Hondt Matthias; Bracke Nathalie; Duchateau Luc; Burvenich Christian; Peremans Kathelijne; De Spiegeleer Bart^*

来源：Malaria Journal, 2013, 12(1): 145.

DOI：10.1186/1475-2875-12-145

• Anti-malaria
• beta-artemether
• Lumefantrine
• Stability-indicating assay
• HPLC-UV
• Fused-core
• Finished pharmaceutical product
• Quality-by-design (QbD)

Background: Artemisinin-based fixed dose combination (FDC) products are recommended by World Health Organization (WHO) as a first-line treatment. However, the current artemisinin FDC products, such as beta-artemether and lumefantrine, are inherently unstable and require controlled distribution and storage conditions, which are not always available in resource-limited settings. Moreover, quality control is hampered by lack of suitable analytical methods. Thus, there is a need for a rapid and simple, but stability-indicating method for the simultaneous assay of beta-artemether and lumefantrine FDC products. %26lt;br%26gt;Methods: Three reversed-phase fused-core HPLC columns (Halo RP-Amide, Halo C18 and Halo Phenyl-hexyl), all thermostated at 30 degrees C, were evaluated. beta-artemether and lumefantrine (unstressed and stressed), and reference-related impurities were injected and chromatographic parameters were assessed. Optimal chromatographic parameters were obtained using Halo RP-Amide column and an isocratic mobile phase composed of acetonitrile and 1mM phosphate buffer pH 3.0 (52:48; V/V) at a flow of 1.0 ml/min and 3 mu l injection volume. Quantification was performed at 210 nm and 335 nm for beta-artemether and for lumefantrine, respectively. In-silico toxicological evaluation of the related impurities was made using Derek Nexus v2.0 (R). %26lt;br%26gt;Results: Both beta-artemether and lumefantrine were separated from each other as well as from the specified and unspecified related impurities including degradants. A complete chromatographic run only took four minutes. Evaluation of the method, including a Plackett-Burman robustness verification within analytical QbD-principles, and real-life samples showed the method is suitable for quantitative assay purposes of both active pharmaceutical ingredients, with a mean recovery relative standard deviation (+/- RSD) of 99.7 % (+/- 0.7%) for beta-artemether and 99.7 % (+/- 0.6%) for lumefantrine. All identified beta-artemether-related impurities were predicted in Derek Nexus v2.0 (R) to have toxicity risks similar to beta-artemether active pharmaceutical ingredient (API) itself. %26lt;br%26gt;Conclusions: A rapid, robust, precise and accurate stability-indicating, quantitative fused-core isocratic HPLC method was developed for simultaneous assay of beta-artemether and lumefantrine. This method can be applied in the routine regulatory quality control of FDC products. The in-silico toxicological investigation using Derek Nexus (R) indicated that the overall toxicity risk for beta-artemether-related impurities is comparable to that of beta-artemether API.

• 出版日期2013-4-30