HPLC测定人血清中薁磺酸钠的浓度

HPLC determination of sodium azulene-sulfonate in human serum

目的:建立高效液相色谱紫外检测法测定血清中薁磺酸钠浓度。方法:采用 Apollo C_(18)色谱柱(5μm,4.6mm×250mm);流动相:乙腈-25mmol·L^(-1)磷酸二氢钾水溶液(pH=4.5;38:62,v/v);流速1mL·min^(-1);进样量20μL;紫外检测波长为293nm;自动进样器温度为室温。采用固相萃取方法对血样进行预处理。结果:线性检测范围为5～500ng·mL^(-1),最低检测浓度为5ng·mL^(-1)。线性良好,r=0.9999。350,200,50,10ng·mL^(-1)4个浓度的相对回收率分别为100.3%,98.8%,99.6%,100.1%;绝对回收率分别为101.6%,101.6%,99.8%,101.3%;内标回收率为100.5%;其日内精密度依次为1.0%,1.6%,2.1%,3.2%;日间精密度依次为2.4%,3.6%,3.2%,5.4%。结论:本法操作简便,灵敏度高,重现性好,可以满足本试验低浓度药物测定及药代动力学、生物利用度研究的需要。

Objective：To establish an HPLC method for measuring the concentration of sodium azulene - sulfonate in the serum. Methods：The HPLC method was used with Apollo C18 （5 μm ,4. 6 mm × 250 mm）column. The mobile phase was composed of acetonitrile and 25 mmol · L- 1KH2 po4 buffer （ pH = 4. 5 ; 38 ： 62, v/v） , the flow rate was 1 mL · min^- 1, the detection wavelength at 293 nm, and column temperature was room temperature. The blood sample was pretreated by solid phase extraction. Results：It showed a good linear range of 5 -500 ng · mL^-1, the minimum detective concentration was 5 ng · mL^-1, r = 0. 9999. The relative recoveries of the four concentrations 350, 200,50, 10 ng · mL^-1 were 100.3%, 98.8%, 99.6%, 100. 1% respectively. The absolute recoveries were 101.6% ,101.6% ,99. 8%, 101.3% respectively. The internal standard recovery was 100. 5%. The intra-day precisions were 1.0%, 1.6% ,2. 1% ,3.2% and the inter - day precisions were 2.4% ,3.6% ,3.2% ,5.4% respectively. Conclusion ：This method is convenient, sensitive and reproducible. It is suitable for determination of low concentration drug and the demand of pharmacokinetic and bioequivalence study.