Validation of high performance liquid chromatography method for determination of meloxicam loaded PEGylated nanocapsules

abstract A method to ensure that an analytical method will produce reliable and interpretable information about the sample must first be validated, making sure that the results can be trusted and traced. In this study, we propose to validate an analytical high performance liquid chromatography (HPLC) method for the quantitation of meloxicam loaded PEGylated nanocapsules(M-PEGNC). We performed a validation study, evaluated parameters including specificity, linearity, quantification limit, detection limit, accuracy, precision and robustness. PEGylated nanocapsules were prepared by interfacial deposition of preformed polymer, and the particle size, polydispersity index, zeta potential, pH value and encapsulation efficiency were characterized. The proposed HPLC method provides selective, linear results in the range of 1.0-40.0 μg/mL; quantification and detection limits were 1.78 μg/mL and 0.59 μg/mL, respectively; relative standard deviation for repeatability was 1.35% and intermediate precision was 0.41% and 0.61% for analyst 1 and analyst 2, respectively; accuracy between 99.23 and 101.79%; robustness between 97.13 and 98.45% for the quantification of M-PEGNC. Mean particle diameters were 261 ± 13 nm and 249 ± 20 nm, polydispersity index was 0.15 ± 0.07 and 0.17 ± 0.06, pH values were 5.0 ± 0.2 and 5.2 ± 0.1, and zeta-potential values were -37.9 ± 3.2 mV e -31.8 ± 2.8 mV for M-PEGNC and placebo(B-PEGNC), respectively. In conclusion, the proposed analytical method is suitable for the quality control of M-PEGNC. Moreover, suspensions showed monomodal size distributions and low polydispersity index indicating high homogeneity of formulations with narrow size distributions, and appropriate pH and zeta potential. The extraction process was efficient for release of meloxicam from nanostructured systems.

摘要 为确保分析方法能够产生关于样品的可靠且可解释的信息，必须首先对该方法进行验证，以确保分析结果可被信赖且可溯源。本研究拟对用于定量测定载美洛昔康聚乙二醇化纳米胶囊（meloxicam loaded PEGylated nanocapsules, M-PEGNC）的高效液相色谱（high performance liquid chromatography, HPLC）分析方法开展验证。本研究完成了方法学验证实验，评估的参数包括专属性、线性、定量限、检测限、准确度、精密度及耐用性。采用预形成聚合物界面沉积法制备聚乙二醇化纳米胶囊，并对其粒径、多分散指数、Zeta电位、pH值以及包封率进行了表征。所建立的HPLC分析方法在1.0~40.0 μg/mL浓度范围内具有良好的选择性与线性关系；定量限与检测限分别为1.78 μg/mL和0.59 μg/mL；两名分析人员的重复性相对标准偏差分别为1.35%，中间精密度分别为0.41%与0.61%；载美洛昔康聚乙二醇化纳米胶囊的定量回收率处于99.23%~101.79%之间，耐用性指标处于97.13%~98.45%之间。载药纳米胶囊（M-PEGNC）与空白纳米胶囊（B-PEGNC）的平均粒径分别为261±13 nm与249±20 nm，多分散指数分别为0.15±0.07与0.17±0.06，pH值分别为5.0±0.2与5.2±0.1，Zeta电位分别为-37.9±3.2 mV与-31.8±2.8 mV。综上，所建立的分析方法适用于载美洛昔康聚乙二醇化纳米胶囊的质量控制。此外，纳米混悬剂呈现单峰粒径分布且多分散指数较低，表明制剂具有较高的均一性与较窄的粒径分布，且pH值与Zeta电位均符合要求。本研究采用的提取方法可高效地从纳米结构载体中释放美洛昔康。