HPLC法测定大鼠血浆中奥拉西坦的浓度

目的:建立大鼠血浆中奥拉西坦浓度的检测方法。方法:取大鼠8只,一次性灌胃给予奥拉西坦200 mg.kg-1,给药2 h后取血,采用高效液相色谱法检测其血药浓度。色谱柱:Symmetry shiedTMRP18;流动相:乙腈-水(3.2∶96.8,V/V),流速:0.8 mL.min-1;检测波长:210 nm;柱温:40℃;进样量:20μL。结果:奥拉西坦的保留时间为3.65 min;其检测浓度的线性范围为2～100 mg.L-1(r=0.999 8);低、中、高浓度血浆样品的绝对回收率分别为(91.1±5.17)%、(97.9±3.22)%、(98.4±1.99)%,方法回收率分别为(96.35±3.01)%、(101.40±3.41)%、(102.23±1.37)%;日内RSD分别为2.86%、4.38%、3.54%,日间RSD分别为3.25%、3.17%、3.14%;奥拉西坦血浆样品-40℃保存30 d及反复冻融3次后性质稳定,血浆样品处理后放置4 h对测定无影响。8只大鼠血浆样品中奥拉西坦的平均浓度为(35.58±10.83)mg.L-1。结论:本方法简便、准确、灵敏度高、重现性好,可用于大鼠血浆中奥拉西坦浓度的测定。

Determination of Oxiracetam.Concentration in Plasma of Rats by HPLC

OBJECTIVE： To establish a method for the determination of oxiracetam concentration in plasma of rats. METHODS： 8 rats were collected and given 200 mg.kg-1 oxiracetam. Blood samples were collected 2 h after medication and determined by HPLC. A Symmetry shiedTM RP18 column was used with mobile phase consisted of acetonitrile-water （3.2 ： 96.8, V/V） at the flow rate of 0.8 mL. min-1. The detection wavelength was set at 210 nm and column temperature was 40 ℃. The injection volume was 20 μL. RESULTS： The retention time of oxiracetam was 3.65 min. The linear range of oxiracetam was 2-100 mg.L-1 （r=0.999 8）. The absolute recoveries of plasma samples at low, medium and high concentrations were （91.1 ± 5.17）%, （97.9 ± 3.22）% and （98.4± 1.99）%, respectively. The method recoveries were （96.35 ±3.01）%, （101.40 ±3.41）% and （102.23 ± 1.37）%, respectively. The intra-day RSD of plasma samples at three concentrations were 2.86%, 4.38% and 3.54%, and the inter-day RSD were 3.25%, 3.17% and 3.14%, respectively. The plasma samples were stable when stored at --40 % for 30 days or freeze-thaw for thrice and processed samples were stable within 4 h. Average concentration of oxiracetam in plasma sample was （35.58 ± 10.83） mg-L-1. CONCLUSION： The method is simple, accurate with high sensitivity and reproducibility. It is fitted for the determination of oxiracetam concentration in plasma of rats.