脑损伤后脑脊液腺苷浓度测定方法的建立

Determination of adenosine concentration in human cerebrospinal fluid after brain injury

目的建立人脑脊液中腺苷(adenosine,AD)浓度的检测方法。方法收集脑损伤后病人脑脊液,采用ODS反相C18柱检测,以50mmol/L磷酸二氢钠∶甲醇(80∶20)为流动相,流速为1.0ml/min,检测波长为254nm。结果该方法在检测0.78～15mg/L腺苷浓度有着良好的线性关系(R2=0.9981),保留时间为4.9min,平均加样回收率为(98.59±0.65)%,日内相对标准偏差小于2.46%,日间相对标准偏差小于3.56%。脑损伤病人脑脊液腺苷含量高于正常腺苷范围。结论本方法具有灵敏度高、特异性强、重复性好、操作简便等优点,可用于脑脊液中腺苷浓度的测定。

Objective To establish a high performance liquid chromatographic （HPLC） method for the determination of adenosine concentration in human cerebrospinal fluid （CSF）. Methods Human CSF samples were collected in patients with brain injuries,and then a reversed phase（RP） ODS C18 column with mobile phase of 50 mmol/L NaH2PO4：methanol （80：20） was used to detect the adenosine concentration in these samples at the flow rate of 1.0 ml/min and wave length of 254 nm. Results The standard curve showed a good linearity range between 0.78 and 15 mg/L（R2 = 0. 9981 ） ,the reserve time was 4.9 min, the average recovery of adenosine was （98.59 ± 0.65 ）% （n = 6）, and the relative standard deviation （RSD） in one day and three days was less than 2.46% and 3. 56%, respectively. Adenosine contents in CSF of patients with brain injuries were higher than that of normal subjects. Conclusion RP - HPLC method is of high sensibility, specificity, good repeatability and easy operation. It can be used for the determination of adenosine concentration in CSF.