HPLC测定大鼠脑脊液中梓醇的含量

HPLC determination of catalpol in cerebrospinal fluid of rats

目的：建立大鼠脑脊液中梓醇的含量测定方法。方法：大鼠按5．0mg·kg^-1尾静脉注射1．0g·L^-1梓醇生理盐水注射液，40min后，经大脑蛛网膜下腔抽取脑脊液；另抽取正常大鼠脑脊液作为空白对照，置-20℃冰箱保存备用。样品经处理后用HPLC检测。选用Hypersil C18反相色谱柱，流动相为水-乙腈（99．5：0．5），流速1．0mL·min^-1，检测波长210姗。结果：脑脊液中梓醇在0．5～40mg·L^-1浓度线性关系良好，r=0．9997；测得低、中、高浓度绝对回收率为（90．2±1．71）％，（89．1±1．17）％，（86．9±0．98）％；方法回收率为（99．8±1．98）％，（101．1±3．04）％，（100．1±2．30）％；日内、日间精密度均小于4％，在-20℃下冷冻保存15d稳定。结论：本方法操作简便，能较准确的测出脑脊液中的梓醇含量。

Objective： To establish an HPLC method for determination of catalpol in CSF （cerebrospinal fluid） of rats. Method： Rats were intravenously injected 1. 0 g ·L^-1 catalpol physiological saline, and the sample of CSF from subarachnoid space of the cerebrum 40 minutes of injection. The sample of CSF from normal rats was used for blank control, the all samples were preserved in a refrigerator of - 20 ℃, and use HPLC was employed to determine the catalpol content. The separation of catalpol was performed on Hypersil C18 reversion phase chromatographic column. The mobile phase consisted of water-acetonitrile （99. 5： 0. 5 ） with a flow rate of 1.0 mL·min^-1 and detection wavelength of 210 nm. Result： The linear range of catalpol in CSF was 0.5-40 mg · L^-1 （r =0. 999 7 ）. The absolute recoveries were （90. 2 ± 1.71 ） % , （89. 1 ±1. 17） % and （86. 9 ± 0. 98） % ; and the methodological recoveries were （99. 8 ± 1.98）%, （ 101.1 ±3. 04）%, （ 100. 1 ±2. 30）% respectively. The within-day and between-day derivation RSD were less than 4%. Catalpel was stable in a refrigerator of -20℃ for 15 days. Conclusion： The method is simple and accurate for the determination of the content of catalpol in CSF.