Boiero Carolina, Allemandi Daniel, Longhi Marcela, Llabot Juan M
Departamento de Farmacia, Facultad de Ciencias Químicas, Universidad Nacional de Córdoba, Córdoba 5000, Argentina.
Departamento de Farmacia, Facultad de Ciencias Químicas, Universidad Nacional de Córdoba, Córdoba 5000, Argentina.
J Pharm Biomed Anal. 2015;111:186-9. doi: 10.1016/j.jpba.2015.03.031. Epub 2015 Apr 3.
An isocratic high-performance liquid chromatographic method was developed and validated for the simultaneous determination of human serum albumin (HSA) and timolol in albumin nanoparticles. This method involved a reversed-phase-C18 column thermostated at 25 °C, UV detection at 276 nm, flow rate of 1.0 ml/min and a mobile phase compounded by 0.05% (v/v) trifluoroacetic acid in water/0.05% (v/v) trifluoroacetic acid in an acetonitrile (40:60, v/v) solution. The elution times for albumin and timolol were 1.84 ± 0.05 min and 2.67 ± 0.04 min, respectively. Calibration curves were linear from 0.2 to 100 mg/ml for HSA and 0.01 to 1 mg/ml for timolol. Limits of quantification were 0.2 mg/ml for HSA and 0.01 mg/ml for timolol. The values of accuracy and precision of intra- and inter-day variation studies were within acceptable limits, according to the US Food and Drug Administration Guidance for Industry. The described method has proved to be useful to give accurate measurements of human serum albumin and timolol from albumin nanoparticles to determine the percentage of encapsulation and the process yield.
建立了一种等度高效液相色谱法并进行了验证，用于同时测定白蛋白纳米粒中的人血清白蛋白（HSA）和噻吗洛尔。该方法采用在25℃恒温的反相C18柱，在276nm处进行紫外检测，流速为1.0ml/min，流动相由0.05%（v/v）三氟乙酸水溶液/0.05%（v/v）三氟乙酸乙腈溶液（40:60，v/v）混合而成。白蛋白和噻吗洛尔的洗脱时间分别为1.84±0.05分钟和2.67±0.04分钟。HSA的校准曲线在0.2至100mg/ml范围内呈线性，噻吗洛尔的校准曲线在0.01至1mg/ml范围内呈线性。HSA的定量限为0.2mg/ml，噻吗洛尔的定量限为0.01mg/ml。根据美国食品药品监督管理局的行业指南，日内和日间变异研究的准确度和精密度值均在可接受范围内。所描述的方法已被证明可用于准确测量白蛋白纳米粒中的人血清白蛋白和噻吗洛尔，以确定包封率和工艺产率。
