Abstract Artemether; a sesquiterpene lactone is widely used for the treatment of malaria as artemisinin-based combination therapy (ACT). The present work involves the development and validation of sensitive reversed-phase high-performance liquid chromatography (RP-HPLC) method for quantification of artemether (ART) in polymeric nanoparticles. ART was transformed to α, β-unsaturated decalones by pre-column acid treatment to enhance the sensitivity of chromophoric group lacking ART for quantification by HPLC-UV. Waters Spherisorb® 5 µm ODS(C18) column (4.6*250 mm) with gradient elution by mobile phase comprising of ACN and PBS (10 mM; pH 6.0) was used to separate acid-treated ART. The analysis was carried at λmax of 253 nm with 20 min and 20 µL run time and injection volume, respectively. The method was found to be linear in the concentration range of 0.5–10 µg mL−1 with 0.09 µg mL−1 and 0.27 µg mL−1 as LOD and LOQ respectively. Further, the method was also found to be specific for ART in presence of blank polymeric nanoparticles, accurate (% average recovery rate 101.7 ± 1.68%), precise (RSD <2%), and robust. The method was successfully used to determine % entrapment efficiency and in vitro release of ART-loaded polymeric nanoparticles with HPLC using a UV-visible detector. Graphical Abstract

中文翻译：

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使用紫外-可见检测器评估柱前酸处理后聚合物纳米颗粒中蒿甲醚的灵敏 RP-HPLC 方法

摘要蒿甲醚；倍半萜内酯作为基于青蒿素的联合疗法 (ACT) 被广泛用于治疗疟疾。目前的工作涉及开发和验证用于定量聚合物纳米粒子中蒿甲醚 (ART) 的灵敏反相高效液相色谱 (RP-HPLC) 方法。通过柱前酸处理将 ART 转化为 α, β-不饱和十酮，以提高缺乏 ART 的发色基团的灵敏度，以便通过 HPLC-UV 进行定量。Waters Spherisorb® 5 µm ODS(C18) 柱（4.6*250 mm）通过包含 ACN 和 PBS（10 mM；pH 6.0）的流动相进行梯度洗脱，用于分离酸处理的 ART。分析在 253 nm 的 λmax 下进行，运行时间和进样体积分别为 20 分钟和 20 µL。发现该方法在 0.5–10 µg mL-1 的浓度范围内呈线性，其中 LOD 和 LOQ 分别为 0.09 µg mL-1 和 0.27 µg mL-1。此外，还发现该方法在空白聚合物纳米粒子的存在下对 ART 具有特异性、准确（% 平均回收率 101.7 ± 1.68%）、精确（RSD <2%）和稳健。该方法已成功用于确定负载 ART 的聚合物纳米粒子的 % 截留率和体外释放，并使用紫外-可见光检测器通过 HPLC。图形概要 该方法已成功用于确定负载 ART 的聚合物纳米粒子的 % 截留率和体外释放，并使用紫外-可见光检测器通过 HPLC。图形概要 该方法已成功用于确定负载 ART 的聚合物纳米粒子的 % 截留率和体外释放，并使用紫外-可见光检测器通过 HPLC。图形概要