高效薄层扫描法测定蛛网膜下腔出血患者脑脊液中血小板活化因子

摘要：建立了测定蛛网膜下腔出血患者（SAH）脑脊液中血小板活化因子（PAF）质量浓度的高效薄层扫描方法。薄层板为高效硅胶G板（100mm×l00mm），展开剂为V（氯仿）:V（甲醇）:V（水）=65:35:6，下层液展开，显色剂为100g/L磷钼酸溶液，展开距90mm，扫描波长630nm。方法的线性范围0.5～2.5μg/L，相关系数0.9990，最小检测质量浓度50ng/L，平均回收率98.6％。运用所建立的方法测定了16例SAH患者发病后脑脊液和10例非神经系统疾病患者脑脊液中PAF的质量浓度及其变化

High Performance Thin Layer Chromatographic (HPTLC) Determination of PAF in Cerebrospinal Fluid of Patients with Subarachnoid Hemorrhage(SAH)

The Platelet Activating Factor (PAF) is believed to be the major function in human after cerebral vascular spasm and cerebral ischemia. PAF has been found to participate in cerebral vascular spasm and cerebral ischemia by the basic and clinical study. The symptom of cerebral vascular spasm and cerebral ischemia has appeared with SAH. It has not been reported that the rule and change of PAF with SAH. In the present work, the concentration of PAF in human cerebral spinal fluid (CSF) with SAH were determined by high performance layer thin chromatography. The TLC plate was coated with high performance silica gel G using V(chloroform):V(methanol):V(water) = 65:35:6 as developing solvent. The PAF was determined by TLC scanning method and detected at 630 nm. The method was applied to determine the concentration of PAF in 16 CSF samples with SAH. The samples were collected in 1-3 d, 7-10 d and 14-21 d. CSF samples were deproteinized with methanol and chloroform. After centrifugation, the chloroform layer separated was dried at room temperature with nitrogen and stored under 20 degrees C in the refrigerator. The linear range of the method was 0.5-2.5 micrograms/L with regression coefficient of 0.9990. The lower limit of detection were 50 ng/L. The recovery of the method was 98.6%. The method enables a simple, rapid and reproducible quantification of PAF with SAH.