离子对高效液相色谱法测定大鼠尿中草酸含量

目的:建立以离子对高效液相色谱法测定大鼠尿中草酸含量。方法:采用色谱柱Symmetry ShieldTM RP18(4.6 mm×250 mm,5μm),保护柱为Symmetry C18(3.9 mm×20 mm,5μm),以乙腈-0.02 mol.L-1磷酸二氢钾溶液(含0.02 mol.L-1四丁基硫酸氢铵,用KOH调pH2.7)(5∶95)为流动相,流速为0.8 mL.min-1,紫外检测波长为220 nm,柱温为40℃。结果:本方法最低检测浓度为5μg.mL-1,尿中草酸高浓度线性范围为67.5～2 160μg.mL-1(r2=0.999 8),低浓度线性范围为6.75～216μg.mL-1(r2=0.997 8),提取回收率>75%,相对回收率为89.2%～105.3%,日内和日间RSD均≤10.2%。结论:本方法简单快速、回收率高、精密度良好,适用于大鼠尿中草酸含量的测定。

Determination of Oxalate in Rat Urine by Ion-pair HPLC

OBJECTIVE： To establish an ion-pair HPLC method for determination of oxalate in rat urine.METHODS： Symmetry ShieldTM RP18（4.6 mm×250 mm,5 μm） column and Symmetry C18（3.9 mm×20 mm,5 μm） guard column were used with acetonitrile-0.02mol·L-1 potassium dihydrogen phosphate solution（containing 0.02 mol·L-1 tetrabutylammonium hydrogensulfate（with pH adjusted to 2.7 using potassium hydroxide）（5∶95） as mobile phase.The flow rate was set at 0.8 mL·min-1;the detection wavelength was set at 220 nm and the column temperature was kept at 40 ℃.RESULTS： The lowest detectable concentration in this method was 5 μg·ml-1.The linear high concentration range of oxalate stood at 67.5-2 160 μg·mL-1（r2=0.999 8） and the linear low concentration range of oxalate stood at 6.75-216 μg·mL-1（r2=0.997 8）.The extraction recovery was75% and the relative recovery was 89.2%-105.3%.Both the intra-day and inter-day RSD were ≤10.2%.CONCLUSION： The method is simple,rapid and precise with high recovery rate,and it is suitable for the determination of oxalate in rat urine.