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获得关于样品的可靠且可解释的信息，首先需对该分析方法进行验证，以保证结果可信且可溯源。本研究针对美洛昔康载聚乙二醇化纳米胶囊（M-PEGNC）的定量分析，提出采用高效液相色谱法（HPLC）进行方法验证。我们开展了验证实验，对专属性、线性、定量限、检测限、准确度、精密度及耐用性等参数进行了评价。采用预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、多分散性指数、Zeta电位、pH值及包封率进行了表征。所建立的HPLC方法具有良好专属性，在1.0~40.0 μg/mL范围内呈良好线性响应；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为1.35%，中间精密度分别为0.41%和0.61%；准确度介于99.23%~101.79%之间；用于M-PEGNC定量分析时的耐用性介于97.13%~98.45%之间。美洛昔康载聚乙二醇化纳米胶囊（M-PEGNC）与空白聚乙二醇化纳米胶囊（B-PEGNC）的平均粒径分别为261±13 nm和249±20 nm，多分散性指数分别为0.15±0.07和0.17±0.06，pH值分别为5.0±0.2和5.2±0.1，Zeta电位分别为-37.9±3.2 mV和-31.8±2.8 mV。综上，所建立的分析方法适用于M-PEGNC的质量控制。此外，纳米混悬剂呈现单峰粒径分布与低多分散性指数，表明制剂均一性高且粒径分布窄，同时具备适宜的pH值与Zeta电位。萃取过程可高效从纳米结构体系中释放美洛昔康。