固相萃取高效液相色谱法测定全血环孢素浓度

Determination of Cyclosporin in Whole Blood by Solid-Phase Extraction and High Performance Liquid Chromatography

目的建立一种简便的测定全血环孢素浓度的高效液相色谱法。方法以VenusilASB C18为色谱柱，流动相为乙腈-水=70：30，柱温70℃，检测波长为210nm，进样量200止；全血样品经10％尿素溶血处理离心后，取上清液直接上PRS—SPE柱，用等体积乙腈-四氢呋喃（80：20）作为洗脱剂，收集洗脱液直接进样测定。结果环孢素全血样品线性范围为30～3500ng·mL-1，低、中、高3种不同浓度（78．2，300．0，1213．0ng·mL-1）方法回收率分别为101．66％，104．06％，100．46％，日内、日间RSD均〈5％。结论该方法准确性好，灵敏度高，样品处理简单，可满足儿科临床中环孢素全血浓度监测需要。

Objective To establish a simple high-performance liquid chromatography （HPLC） method to determinate cyclosporin in whole blood. Methods The HPLC was carried out by using Venusil ASB C18 column, acetonitrile/water（70 ： 30）as a mobile phase, setting column temperature at 70 ℃, detection wavelength at 210 nm. Whole blood samples were centrifugated after treated with 10% urea for hemolysis. Two hundred microliters of the supernatant was directly injected into the PRS - SPE column, using equal volume of acetonitrile/tetrahydrofuran（ 80 ： 20） as the eluent. Results The method is linear at 30 to 3 500 ng· mL-1 curve range. The recoveries at low, medium and high concentrations （78.2, 300.0, 1 213.0 ng ·mL-1 ） were 101.66%, 104.06% and 100.46%, respectively. The intra- and inter-day RSD were less than 5%. Conclusion The method is accurate, sensitive, and the sample is easy to handle, fully meeting the requirement of pediatric clinical practice for determination of cyclosporin in whole blood.