反相高效液相色谱法测定丙泊酚血药浓度

Determination of concentration of propofol in plasma by RP-HPLC method

目的 :建立测定丙泊酚人血药浓度的反相高效液相色谱 荧光检测分析方法。方法 :使用Agi lent 110 0型高效液相色谱仪、PhenomenexSecurityGuard ( 4mm× 3mm )保护柱、DiamonsilC18( 15 0mm× 4 .6mm ,5 μm)色谱柱。在血样中加入麝香草酚 (内标 )的甲醇溶液沉淀蛋白后 ,离心取上清液 ,进行HPLC分析。流动相 :乙腈∶水 =5 6∶4 4(V/V) ;流速 :1.5mL·min- 1;荧光检测 :λex=2 76nm ,λem=312nm。结果 :本法标准曲线线性方程为Y =1.787C - 0 .0 31(r =0 .9997,n =9,线性浓度范围为 0 .0 2～ 8.0mg·L- 1) ;最低检测浓度为 4 μg·L- 1(S/N =2 ) ;平均方法回收率为 98.71%;日内、日间精密度RSD( %)均小于 4 .2 %。结论 :本方法简便、灵敏、准确 ,可用于临床上丙泊酚的血药浓度监测。

AIM:To establish a reversed-phase HPLC method with fluorescence detection for determination of the concentration of propofol in human plasma. METHODS: Agilent 1100 HPLC instrument was used with the column : Diamonsil C 18(150 mm×4.6 mm,5 μm), Phenomenex Security Guard column(4 mm×3 mm). Plasma samples were precipitated with the thymol (internal standard) methanol solution. 20 μL of supernatant fluid was injected for HPLC analysis. The mobile phase was composed of acetonitrile-water(56:44,V/V).The flow rate was 1.5 mL·min -1. The fluorescence detective wavelengths were: λ ex=276 nm, λ em=312 nm. RESULTS: The standard curve equation was Y= 1.787C- 0.031 (r= 0.9997,n=9).The linear range was (0.02—8.0) mg·L -1. The average recovery was 98.71 %.Intra- and inter-day precision(RSD%) were all less than 4.2 %. The limit of detection was 4 μg·L -1(S/N=2). CONCLUSION:The RP-HPLC method is simple, sensitive and accurate for monitoring propofol concentration in clinical plasma samples.