血液、尿液中氯胺酮及其代谢物去甲氯胺酮的HPLC分析

目的 建立血液、尿液中氯胺酮及其代谢物去甲氯胺酮的高效液相色谱(HPLC)分析方法.方法 以非那西丁为内标,检材加入10%的氢氧化钠溶液调节pH值为14,用甲苯提取,离心后取有机层,水浴下吹干,乙腈定容后进HPLC仪分析.结果 检测血液中氯胺酮和去甲氯胺酮的线性范围均是0.05～10μg/mL(r2＞0.999 3),检测尿液中氯胺酮和去甲氯胺酮的线性范围均是0.01～50 μg/mL(r2＞0.999 5).氯胺酮和去甲氯胺酮在血液和尿液中的检测限分别是0.006 μg/mL和0.003 μg/mL.血液和尿液中氯胺酮和去甲氯胺酮的回收率不低于82.4%.检测血液和尿液中氯胺酮和去甲氯胺酮的日内精密度和日间精密度均小于10.0%.将所建的方法应用于给大鼠氯胺酮后的血液和尿液中的氯胺酮和去甲氯胺酮的测定,得到了氯胺酮和去甲氯胺酮在大鼠的药时曲线和尿排药速率曲线. 结论本方法简便、快捷,适用于血液、尿液中氯胺酮及其代谢物去甲氯胺酮的分析.

Determination of ketamine and norketamine in blood and urine by high performance liquid chromatography

OBJECTIVE: To develop a high-performance liquid chromatographic method for determination of ketamine and norketamine in blood and urine. METHODS: The compounds were extracted from blood or urine by liquid-liquid extraction using toluene after blood or urine was adjusted pH to 14. The extracts were analyzed by HPLC. RESULTS: Linear limits of ketamine and norketamine determination in blood ranged from 0.05 microg/mL to 10 microg/mL (R2 > 0.9993) and in urine ranged from 0.01 microg/mL to 200 microg/mL (R2 > 0.9995). Limits of detection (LODs) for ketamine and norketamine were 0.006 microg/mL and 0.003 microg/mL (S/N > or = 3), respectively. The mean extraction recovery was over 82.4% and its coefficients of variation were less than 10.0% for ketamine and norketamine. Concentration-time curves and urinary drug velocity curves of ketamine norketamine were obtained by determinations of them in blood and urine in rat using the developed method. CONCLUSION: The method is sensitive, simple, rapid and suitable for determination of ketamine and norketamine in blood and urine for toxicological and clinical pharmaceutical analysis.