摘要
目的建立利用HPLC法测定荷瘤小鼠组织中马钱子碱浓度的方法。方法以石杉碱甲为内标,采用液-液2次萃取处理小鼠肿瘤、肝、肾、脑、血浆样品。采用Kromasil C18(250 mm×4.6 mm,5μm)和汉邦C18预柱(10 mm×4.6 mm,5μm);流动相:乙腈∶0.01 mol.L-1庚烷磺酸钠与0.02 mol/L磷酸二氢钾的等量混合液(用10%磷酸调pH 2.8)(24∶76);检测波长:264 nm;流速:1.0 mL/min;柱温:35℃。结果内源性杂质对样品测定无干扰。马钱子碱在50～2000 ng/mL范围内线性关系良好。组织和血浆中马钱子碱的绝对回收率为70.23%～99.18%,方法回收率为92.57～108.86%,日内RSD≤9.31%,日间RSD≤8.58%。样品冻融与复溶后稳定性良好。结论该法快速、灵敏、准确,可用于评价马钱子碱制剂在荷瘤小鼠体内的靶向性分布结果。
Objective To establish the HPLC method for determining brucine concentration in the tissues of tumor- bearing mice. Methods The tissue samples of liver, kidney, brain, tumor and plasma were treated with simple liq- uid-liquid extraction before HPLC. Chromatographic separation was achieved on a Kromasil C18 analytical column （250 mm × 4.6 mm, 5 μm） coupled with a C18 guard cartridge （10 mm × 4.6 mm, 5 μm） , maintained at 35℃. The mobile phase consisted of 24 % acetonitrile and 76 % buffer （isometric mixture of 0.01 mol/L sodium heptane sulfonate and 0.02 mol/L potassium dihydrogen phosphate, the pH value was adjusted to 2.8 with 10 % phosphoric acid）. The prepared mobile phase was delivered at a flow rate of 1.0 mL/min. The detection wavelength was set at 264 nm. Results There were no interference peaks co-eluting with the compounds of interest. The extraction recoveries varied from 70.23 % to 99.18 %. The linear range was 50-2000 ng/mL. The intra-day precision was less than 9.31% and inter-day precision was less than 8.58 %. The stability of biosamples was satisfactory. Conclusion The method which is simple, practical and accurate can be applied to the study of targeting distribution of brucine preparations.
出处
《中药新药与临床药理》
CAS
CSCD
北大核心
2011年第4期429-432,共4页
Traditional Chinese Drug Research and Clinical Pharmacology
基金
国家自然科学基金项目(30701111)
