Furman Christophe, Carpentier Rodolphe, Barczyk Amélie, Chavatte Philippe, Betbeder Didier, Lipka Emmanuelle
Inserm, U995-LIRIC, Lille, France.
Faculté de Pharmacie, Plateforme de Binding, Université de Lille, U995-LIRIC, Lille, France.
Electrophoresis. 2017 Oct;38(19):2536-2541. doi: 10.1002/elps.201600552. Epub 2017 Apr 28.
A reversed-phase high-performance liquid chromatography (RP-HPLC) method has been developed and validated for the quantification of paclitaxel encapsulated in biodegradable poly(lactic-co-glycolic) (PLGA) copolymer nanoparticles. This simple (isocratic mode, without additive) and rapid (retention time of the paclitaxel under 4 min) methodology permits the detection of low quantities of paclitaxel in nanoparticulate formulations and the determination of the encapsulation efficiency (EE). Analysis was achieved on an octadecyl stationary phase. The isocratic mobile phase consisted of acetonitrile:water 80:20 (v/v) (flow rate = 0.8 mL/min). Stability of free paclitaxel was preliminary studied in those chromatographic conditions. The calibration curve was linear in the concentration range of 2-10 μg/mL (R = 0.9994). The method was specific with valuable trueness, repeatability (intra-day precision) and intermediate precision (inter-day precision) based on relative standard deviation (RSD) values (less than 2%). The limits of detection (LOD) and quantification (LOQ) were 0.56 and 1.85 ng/mL, respectively. This developed method was successfully employed for quantifying paclitaxel in PLGA 50:50 co-polymer nanoparticles. The accurate knowledge of the encapsulated paclitaxel concentration is essential to define the quantities of PLGA nanoparticles necessary to achieve the in vitro cell viability study.
已开发并验证了一种反相高效液相色谱（RP-HPLC）方法，用于定量测定包裹在可生物降解聚（乳酸-共-乙醇酸）（PLGA）共聚物纳米颗粒中的紫杉醇。这种简单（等度洗脱模式，无添加剂）且快速（紫杉醇保留时间在4分钟以内）的方法能够检测纳米颗粒制剂中低含量的紫杉醇，并测定包封率（EE）。分析在十八烷基固定相上进行。等度流动相由乙腈:水80:20（v/v）组成（流速 = 0.8 mL/min）。在这些色谱条件下初步研究了游离紫杉醇的稳定性。校准曲线在2-10 μg/mL浓度范围内呈线性（R = 0.9994）。基于相对标准偏差（RSD）值（小于2%），该方法具有良好的准确性、重复性（日内精密度）和中间精密度（日间精密度）。检测限（LOD）和定量限（LOQ）分别为0.56和1.85 ng/mL。所开发的方法成功用于定量PLGA 50:50共聚物纳米颗粒中的紫杉醇。准确了解包封的紫杉醇浓度对于确定实现体外细胞活力研究所必需的PLGA纳米颗粒数量至关重要。
