Université de Lyon, Malaria Research Unit, ICBMS, UMR 5246 CNRS-INSA-CPE-Université Lyon1, 69622 Villeurbanne, France - Groupement Hospitalier Nord, Institut de Parasitologie et Mycologie Médicale, Hospices Civils de Lyon, 69004 Lyon, France.
Université de Lyon, Malaria Research Unit, ICBMS, UMR 5246 CNRS-INSA-CPE-Université Lyon1, 69622 Villeurbanne, France.
Parasite. 2022;29:18. doi: 10.1051/parasite/2022019. Epub 2022 Mar 29.
Artesunate is the current most potent antimalarial drug widely used for the treatment of malaria. Considering the emergence of artemisinin resistance, several situations may require a simple method for artesunate quantification. We thus developed a quantitative and a semi-quantitative biological method for the determination of artesunate in liquid samples. The tests are based on the measurement of samples' antimalarial activity on Plasmodium falciparum 3D7 using a modified SYBR Green I drug susceptibility test. For the quantitative test, we established a standard curve that resulted from a dose-response curve and evaluated its performances using controls samples. Whereas the linear regression analysis between artesunate concentration and antimalarial activity showed promising results (linearity range 1.5-24.6 ng/mL, r = 0.9373), we found that artesunate content of the controls was significantly overestimated (p = 0.0313). For the semi-quantitative test, we compared the antimalarial activities of samples collected during permeation studies of artesunate to that of a reference (artesunate IC) by statistical analysis. We demonstrated that antimalarial activities of samples from permeation tests using a powder formulation of artesunate were greater than those of samples from tests using a solution formulation. Bioassays can be simple techniques to assess artesunate in liquid samples, particularly in resource-limited settings. Comparison with reference methods is still recommended when accurate drug quantification is required.
青蒿琥酯是目前最有效的抗疟药物，广泛用于治疗疟疾。鉴于青蒿素耐药性的出现，在某些情况下可能需要一种简单的青蒿琥酯定量方法。因此，我们开发了一种定量和半定量的生物方法，用于测定液体样本中的青蒿琥酯。这些测试基于用改良的 SYBR Green I 药物敏感性试验测量样本对恶性疟原虫 3D7 的抗疟活性。对于定量测试，我们建立了一个标准曲线，该曲线来自剂量反应曲线，并使用对照样品评估其性能。虽然青蒿琥酯浓度与抗疟活性之间的线性回归分析显示出有希望的结果（线性范围 1.5-24.6ng/mL，r=0.9373），但我们发现对照品中的青蒿琥酯含量被显著高估（p=0.0313）。对于半定量测试，我们通过统计分析比较了青蒿琥酯渗透研究中收集的样本与参考品（青蒿琥酯 IC）的抗疟活性。我们证明，使用青蒿琥酯粉末制剂进行渗透试验的样本的抗疟活性大于使用溶液制剂进行试验的样本的抗疟活性。生物测定可以是评估液体样本中青蒿琥酯的简单技术，特别是在资源有限的环境中。当需要准确的药物定量时，仍然建议与参考方法进行比较。
