高效液相色谱-荧光法检测脑脊液的异丙酚浓度

目的：建立测定脑脊液异丙酚浓度的方法。方法：应用反相高效液相色谱技术，脑脊液样品经高速离心（12500r／min）3min，0．45μm滤膜过滤，以甲醇-水（80：20）流动相，流速1ml／min，柱温30℃，进样量20μl。经Supelco Discovery C18柱分离后，荧光检测，λex=276nm，λem=310nm。结果：脑脊液异丙酚浓度测定线性范围为5～200ng／ml（r=0．9994）。高、中、低3个浓度的回收率分别为101．29／6、99．8％、98．8％；日间RSD分别为1．55％、1．73％、6．01％；日内RSD分别为1．69％、2．37％、8．60％。最低检测浓度为2ng／ml。结论：该方法快速、准确、灵敏，可用于脑脊液异丙酚浓度的测定。

Determination of propofol in cerebral spinal fluid by HPLC with fluorescence detection

OBJECTIVE: To establish a method for determining propofol in human cerebrospinal fluid (CSF). METHODS: Reverse phase high-performance liquid chromatography (HPLC) with fluorescence detection was applied to quantitative analysis. CSF samples were centrifuged (12,500 r/min for 3 min) and filtered (the diameter of the filter is 0.45 microm). Twenty mul of supernatant was directly injected and separated by Supelco Discovery C(18)column. The mobile phase was composed of methanol-water (80:20); the flow rate was 1 ml/min, and the column temperature was 30 degree. The fluorescence detective waves were: lambda ex=276 nm and lambda em=310 nm. RESULTS: The linear range of propofol in CSF was 5-200 ng/ml (r=0.9994). The recovery rates for high, intermediate and low concentrations were 101.2%, 99.8%, 98.8%, respectively. The RSD of inter-day assay was 1.55%, 1.73%, 6.01% and it of intra-day assay was 1.69%, 2.37%, 8.60%. The limit of detection proved to be 2 ng/ml. CONCLUSION: The method is rapid, simple, accurate and sensitive for measurement of propofol concentration in CSF.