National Center for Safety Evaluation of Drugs, National Institutes for Food and Drug Control, A8 Hong Da Zhong Lu, Beijing Economic-Technological Development Area, Beijing 100176, China.
J Chromatogr B Analyt Technol Biomed Life Sci. 2011 Mar 1;879(7-8):548-52. doi: 10.1016/j.jchromb.2011.01.012. Epub 2011 Jan 15.
A sensitive and selective liquid chromatographic-tandem mass spectrometric method was developed for quantification of triamcinolone acetonide (TA) in rabbit ocular tissues. After a simple liquid-liquid extraction using tert-butyl methyl ether, TA and internal standard methylprednisolone were separated on a C₁₈ column with a mobile phase of acetonitrile:water:formic acid (60:40:0.1, v/v/v) and quantified by the use of selected reaction monitoring mode with a total run time of 4 min. The method was validated in tissue homogenates with a daily working range of 1-1000 ng/mL with correlation coefficient of more than 0.99 and a sensitivity of 1 ng/mL as lower limit of quantification, respectively. The mean intra-day and inter-day precisions were less than 10% and accuracy values were higher than 90%. This method was fully validated for the accuracy, precision and stability studies. The method proved to be accurate and specific, and was applied to an in vivo biodistribution study of TA after intravitreal injection to rabbits. Values of mean residence time in vitreous humor, crystalline lens and aqueous humor were 27.7, 35.8 and 20.0 days, respectively.
建立了一种灵敏、选择性的液相色谱-串联质谱法，用于兔眼组织中曲安奈德（TA）的定量分析。采用叔丁基甲醚进行简单的液液萃取后，TA 和内标甲基强的松龙在 C₁₈ 柱上用乙腈：水：甲酸（60:40:0.1，v/v/v）作为流动相分离，采用选择反应监测模式定量，总运行时间为 4 分钟。该方法在组织匀浆中的验证结果表明，每日工作范围为 1-1000ng/mL，相关系数大于 0.99，灵敏度为 1ng/mL，为定量下限。日内和日间精密度均小于 10%，准确度值均高于 90%。该方法经过了准确性、精密度和稳定性研究的全面验证。该方法准确、特异，应用于兔眼内注射 TA 的体内生物分布研究。玻璃体液、晶状体和房水中的平均驻留时间分别为 27.7、35.8 和 20.0 天。
