Teixeira Maribel, Afonso Carlos M M, Pinto Madalena M M M, Barbosa Carlos Maurício
Centro de Estudos de Química Orgânica, Fitoquímica e Farmacologia da Universidade do Porto-Faculdade de Farmácia do Porto, R. Aníbal Cunha 164, 4050 Porto, Portugal.
J Chromatogr Sci. 2003 Aug;41(7):371-6. doi: 10.1093/chromsci/41.7.371.
This work relates the development and validation of a simple reversed-phase high-performance liquid chromatographic (HPLC) method for the analysis of xanthone (XAN) and 3-methoxyxanthone (3-MeOXAN) in poly(D,L-lactide-co-glycolide) (PLGA) nanocapsule formulations. This method does not require any complex sample extraction procedure. Chromatographic separation is made with a reversed-phase C(18) column, using methanol-water (90:10, v/v) 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 min of chromatographic time. The developed method is statistically validated according to United States Pharmacopoeia 25 and International Conference on Harmonization guidelines for its specificity, linearity, accuracy, and precision. The assay method proposed in this study is specific for XAN and 3-MeOXAN in the presence of nanocapsule excipients. Diode-array analyses confirm the homogeneity of XAN and 3-MeOXAN peaks in stressed conditions. Standard curves are linear (r > 0.999) over the concentration range of 0.4-2.5 and 1.0-5.8 micro g/mL for XAN and 3-MeOXAN, respectively. Recovery from nanocapsules ranges from 99.6% to 102.8% for XAN and 98.8% to 102.4% for 3-MeOXAN. Repeatability (intra-assay precision) is acceptable with relative standard deviation values of 1.2% for XAN and 0.3% for 3-MeOXAN.
本研究建立并验证了一种简单的反相高效液相色谱（HPLC）法，用于分析聚（D，L-丙交酯-共-乙交酯）（PLGA）纳米胶囊制剂中的呫吨酮（XAN）和3-甲氧基呫吨酮（3-MeOXAN）。该方法无需任何复杂的样品提取程序。采用反相C(18)柱进行色谱分离，以甲醇-水（90:10，v/v）为流动相，流速为1 mL/min。通过237 nm处的紫外检测进行鉴定。等度洗脱系统在室温下运行，色谱分析时间为7分钟。根据美国药典25版和国际协调会议指南，对所建立的方法进行统计学验证，考察其特异性、线性、准确性和精密度。本研究提出的测定方法对纳米胶囊辅料存在下的XAN和3-MeOXAN具有特异性。二极管阵列分析证实了在强制条件下XAN和3-MeOXAN峰的均一性。XAN和3-MeOXAN的标准曲线在浓度范围分别为0.4 - 2.5和1.0 - 5.8μg/mL时呈线性（r > 0.999）。XAN从纳米胶囊中的回收率为99.6%至102.8%，3-MeOXAN为98.8%至102.4%。重复性（批内精密度）良好，XAN的相对标准偏差值为1.2%，3-MeOXAN为0.3%。
