Teixeira Maribel, Afonso Carlos M M, Pinto Madalena M M, Barbosa Carlos Maurício
Centro de Estudos de Química Orgânica, Fitoquímica e Farmacologia da Universidade do Porto e Laboratório de Química Orgânica-Faculdade de Farmácia do Porto, R. Aníbal Cunha, 164, 4050-047 Porto, Portugal.
J Chromatogr Sci. 2008 Jul;46(6):472-8. doi: 10.1093/chromsci/46.6.472.
A rapid and simple high-performance liquid chromatographic method for the analysis of 1,3-dihydroxy-2-methylxanthone (DHMXAN) in biodegradable poly(D,L-lactide-co-glycolide) (PLGA) nanosphere and nanocapsule formulations is developed and validated. The method does not require any complex sample extraction procedure. Chromatographic separation is made with a reversed-phase C18 column, using methanol-water (90:10, v/v) containing 1% (v/v) acetic acid as a mobile phase at a flow rate of 1 mL/min. Identification is made by UV detection at 237 nm. The isocratic system operates at ambient temperature and requires 7.5 min of chromatographic time. The developed method is statistically validated according to ICH guidelines and USP 29 for its specificity, linearity, accuracy, and precision. The assay method proposed in this study is specific for DHMXAN in the presence of nanosphere and nanocapsule excipients. Diode-array analyses confirm the purity of DHMXAN peak in stress conditions (> 99.0%). The method is shown to be linear (r > or = 0.999) over the concentration range of 0.25-3.0 microg/mL. Recovery ranges from 99.0% to 102.7% (RSD: 1.49%) and from 98.3% to 101.6% (RSD: 1.07%) for nanospheres and nanocapsules, respectively. Repeatability (intra-assay precision) and intermediate precision is acceptable with RSD values ranging from 0.6% to 1.9% and from 0.3% to 2.0%, respectively. The method is shown to be suitable for the evaluation of DHMXAN content entrapped in PLGA nanoparticles.
开发并验证了一种快速简便的高效液相色谱法，用于分析可生物降解的聚（D，L-丙交酯-共-乙交酯）（PLGA）纳米球和纳米囊制剂中的1,3-二羟基-2-甲基呫吨酮（DHMXAN）。该方法不需要任何复杂的样品提取程序。采用反相C18柱进行色谱分离，以含1%（v/v）乙酸的甲醇-水（90:10，v/v）为流动相，流速为1 mL/min。通过在237 nm处进行紫外检测进行鉴定。等度系统在室温下运行，色谱时间为7.5分钟。根据ICH指南和USP 29对所开发方法的特异性、线性、准确性和精密度进行了统计学验证。本研究中提出的测定方法对纳米球和纳米囊辅料存在下的DHMXAN具有特异性。二极管阵列分析证实了应激条件下DHMXAN峰的纯度（>99.0%）。该方法在0.25-3.0μg/mL的浓度范围内呈线性（r≥0.999）。纳米球和纳米囊的回收率分别为99.0%至102.7%（RSD：1.49%）和98.3%至101.6%（RSD：1.07%）。重复性（批内精密度）和中间精密度均可接受，RSD值分别为0.6%至1.9%和0.3%至2.0%。该方法被证明适用于评估PLGA纳米粒中包裹的DHMXAN含量。
