HPLC法测定全血中环孢素浓度

Measurement of ciclosporin in whole blood by HPLC

目的建立测定全血中环孢素（CsA）浓度的固相萃取HPLC法。方法全血样品用硫酸锌甲醇混合液处理，取上清液进样C18（10mm×3.9mm）柱，经水冲洗去杂质后，用纯甲醇洗脱CsA，进入C18（250mm×4．2mm）柱，以流动相：乙腈-甲醇-水（60：10：30，V／V／V）洗脱后进行定量测定。结果本法最低检出浓度为40μ·L^-1，线性范围为62．5—2000μ·L^-1，方法回收率为90％-95％，提取回收率〉80％（n=5），日内RSD为3．25％-4．12％，日间RSD为3．37％-6．87％。结论本法测定CsA前处理步骤简便，结果可靠，可用于临床标本检测。

A/M To establish a HPLC method for determination of ciclosporin. METHODS Blood samples（ 1 mL） were deproteinzed by addition of 1 mL of a mixture of 800 mL of methanol and 200 mL of aqueous zinc sulfate solution （0.4 mmol.L^-1） .Then the supernatant liquid was pipetted into C18（10 mm× 3.9 mm） column.First,samples were injected into the HPLC system and transferred through the extraction column by water 2.0 mL. min^-1 for 2 min; second, the ciclosporin was eluted by methanol,The separation was achieved using an C18（250mm × 4.2mm） column with a mo- bile phase of acetonitrile-methanol-water（60： 10：30, V/V/V）, then UV detection was performed at 210 nm. RESULTS In whole blood the hner range was 62.5 - 2 000μg.L^- 1, the minimal detectable concentration of this method was 40 μg. L^- 1,the precision of method was 90% -95% ,the extraction recovery of method was more than 80% （n = 5）, the RSD was within 3.25 % - 4.12 % within day, and within 3,37 % - 6.87 % between days. CONCLUSION The method is simple and could be applied to examine the clinical samples.