高效液相色谱法测定血浆中氯胺酮对映体的浓度

目的:建立以高效液相色谱手性固定相法测定血浆中氯胺酮对映体浓度的方法。方法:色谱柱为Chirobiotic V手性柱,流动相为甲醇-冰醋酸-三乙胺(100∶0.02∶0.02),检测波长为269nm,流速为1mL·min-1,内标为非那西丁。结果:S-氯胺酮、R-氯胺酮分别在19.5～2600(r=0.9992)、26～2600ng·mL-1(r=0.9989)范围内线性关系良好,最低检测浓度分别为13.0、19.5ng·mL-1;相对回收率分别为99.48%～103.40%、98.53%～103.20%,绝对回收率分别为63.2%～73.1%、64.5%～72.3%;日内RSD分别为3.3%～6.4%、3.5%～5.7%,日间RSD分别为7.1%～9.7%、6.3%～9.9%。结论:本方法简便、准确、灵敏、快速、专属,可用于血浆中R-氯胺酮和S-氯胺酮的测定及药动学研究。

Determination of Ketamine Enantiomers in Plasma by HPLC

OBJECTIVE： To establish an HPLC-CSP （chiral stationary phase） method for the determination of plasma levels of ketamine enantiomers. METHODS： S-ketamine and R-ketamine were separated on Chirobiotic V chiral column using mobile phase consisted of methanol-acetic acid-triethylamine （100 ∶ 0.02 ∶ 0.02） with phenacetin as internal standard. Flow rate was set at 1 mL·min-1 and detection wavelength was 269 nm. RESULTS： The calibration curve was linear in the range of 26～2 600 ng·mL-1 for R-ketamine （r=0.998 9） and 19.5～2 600 ng·mL-1 for S-ketamine（r=0.999 2）.The detection limits were 19.5 ng·mL-1 for R-ketamine and 13.0 ng·mL-1 for S-ketamine. The relative recovery of R-ketamine was 98.53%～103.20% and that of S-ketamine was 99.48%～103.40%. The absolute recovery were 64.5%～72.3% for R-ketamine and 63.2%～73.1% for S-ketamine. The intra-day and inter-day of RSD were 3.5%～5.7% and 6.3%～9.9% for R-ketamine and 3.3%～6.4% and 7.1%～9.7% for S-ketamine, respectively. CONCLUSION： This method is simple, accurate, sensitive, rapid and specific for the determination and pharmacokinetic study of S-ketamine and R-ketamine in plasma.