摘要
目的:建立HPLC柱后修饰荧光检测法测定大鼠血浆中氢溴酸加兰他敏的方法。方法:血浆样本经甲醇直接沉淀蛋白后采用柱后修饰荧光检测法测定。色谱柱为Hypersil ODS2柱(250mm×4.6mm,5.0μm);流动相为0.1%三乙胺溶液-甲醇(60∶40),柱温30℃;流速0.8mL·min-1,进样量20μL;荧光检测的激发波长和发射波长分别为290nm和320nm。柱后修饰采用25mmol·L-1磷酸二氢钠缓冲溶液(磷酸调节溶液pH值至3.8);流速0.2mL·min-1。结果:氢溴酸加兰他敏在3.23～828.13ng·mL-1内线性关系良好(r=0.999 8),最低定量限为3.23ng·mL-1。提取回收率为84.32%～95.24%,日内精密度为2.67%～5.46%,日间精密度为3.10%～7.48%。结论:柱后修饰较大提高了氢溴酸加兰他敏检测灵敏度,该方法简便、准确、所需血浆量少,为临床进一步研究氢溴酸加兰他敏提供了基础。
OBJECTIVE To establish a HPLC with post-column modification and fluorescence detection for the determination of galanlamine hydrobromide in rat plasma. METHODS The sample was added with melhanol to precipitate protein and ana lyzed with post-column modification and fluorescence detection. All separations were carried out on a Hypersil ODS2 column （250 mm× 4. 6 mm i. d. with 5. 0μm particle size）. The mobile phase was composed of acetonitrile：0.1% triethylaminc solu tion （60：40）. The solvent flow rate was 0. 8 mL.min ^-1, the injection volume was 20 μL, and the column temperature was maintained at 30 ℃. The excitation and emission wavelengths were 290 and 320 nm, respectively. Posl-column modification was achieved by buffer solution （25 mmol.L ^-1 sodium dihydrogen phosphate adjusted to pH 3.8 with phosphoric acid）. The flow raze was 0. 2 mL,min t. RESULTS The good iinearity of galantamine hydrobromidc was obtained in the range of 3. 23 - 828. 13 ng ·mL^-1. The correlation coefficient was 0. 999 8 and the lower limit of quantification was 3.23 ng-mL^-1. The extrac tion recoveries were in the range of 84. 32% - 95.24%. The precision of intra batch and inter batch were 2. 67- 5. 46% and 3. 10% - 7. 48%, respectively. CONCLUSION Post-column modification was proved to be an effective way for enhancing the sensitivity of detection of galantamine hydrobromide. The method is simple, accurate and provides an experimental basis for the development of research of galantamine hydrobromide in medicine.
出处
《中国医院药学杂志》
CAS
CSCD
北大核心
2012年第24期1968-1971,共4页
Chinese Journal of Hospital Pharmacy
