用HPLC分析脑梗塞病人脑脊液中LEK和一个未知组分

HPLC determination of LEK and an unknown fraction in CSF of patients with cerebrel infarction.

本实验建立了HPLC配备荧光检测器(FD)梯度洗脱、检测CSF中丹磺酰亮氨酸脑啡肽(DNS-LEK)的方法。在测定LEK的同时,从CSF中发现一个响应值高、保留时间恒定的与脑梗塞有关的未知组分(unknown fraction,Fu),并对它进行了分离纯化和初步的化学鉴定。发现脑梗塞病人CSF中LEK的含量(128.67±42.76ng/ml)明显高于对照值(78.25±16.09 ng/ml),(P<0.01);Fu的含量(峰高值为0.54471±0.23277 mm)显著低于对照值(峰高值为0.78472±0.24940mm)(P<0.001)。脑梗塞中LEK含量增高,而Fu则减少,两者在缺血发作中起何作用,都是有待继续研究的问题。

In our-work HPLC-FD assay for eluating and detecting the pre-column derivatives DNS-LEK was developed and evaluated, An unknown fraction (FU) related to cerebral infarction in CSF was found by this method simultaneously. The mean level (peak height) of FU in human CSF is 20 times the size of noise. Preliminary chemical identification showed that FU is a simple substance other than amino acid or peptide, and can not be hydrolyzed by HCI. Molecular weight of FU is 228 determined by mass spectrometry. HPLC retention time of FU is different from that of monoamine and their metabolites. Both the retention time and peak height or area of FU are reproducible. The result showed that the LEK level of CSF in patients with cerebral infarction is significantly higher (128.67±42.76ng/ml) than that of controls (78.25 = 16.09ng/ml), and FU level is significantly lower (0.54471±0.23277 mm) than that of controls (0.78472±0.24940mm). The role of these changes in the occurrence of cerebral infraction remains unknown and needs further research.