Frölich L, Dirr A, Götz M E, Gsell W, Reichmann H, Riederer P, Maurer K
Department of Psychiatry I, University of Frankfurt am Main, Federal Republic of Germany.
J Neural Transm (Vienna). 1998;105(8-9):961-73. doi: 10.1007/s007020050105.
Four different methods to measure acetylcholine (ACh) and choline (Ch) concentration, i.e. thermospray/mass spectroscopy (TS/MS), high pressure liquid chromatography/mass spectroscopy (HPLC/MS), high pressure liquid chromatography/electrochemical detection (HPLC/ECD) and gas chromatography/mass spectroscopy (GC/MS), both latter methods coupled to a solid phase extraction system were compared for their applicability to human lumbar cerebrospinal fluid (CSF). Furthermore, samples from 15 control persons and 11 patients with dementia of Alzheimer-type (DAT) were compared to search for an ACh deficit in the CSF in DAT. GC/MS was the most sensitive, but most laborious method, and HPLC/ECD was acceptably sensitive, reliable and more specific. TS/MS was not specific enough for CSF extracts and HPLC/MS was more specific, but far less sensitive. Thus, only GC/MS and HPLC/ECD were qualified to detect ACh in human CSF extracts. Comparison of GC/MS and HPLC/ECD revealed highly correlated levels of ACh (r = 0.999). Using HPLC/ECD, ACh concentrations were greatly reduced in the DAT group (3.75 +/- 1.40 pmol/ml CSF) as compared to the controls (6.14 +/- 1.39 pmol/ml CSF), but the difference between controls and DAT patients was not statistically significant due to the number of samples below detection limit (8 out of 11 samples in DAT, 7 out of 15 in controls). Ch concentrations were not statistically significant between the two groups. The data show that methodological limitations preclude a widespread clinical application of determining ACh in the human CSF. Despite of reductions of ACh in the CSF in DAT, the determination of Ach in the CSF is not suitable for diagnostic purposes.
比较了四种测量乙酰胆碱（ACh）和胆碱（Ch）浓度的不同方法，即热喷雾/质谱法（TS/MS）、高压液相色谱/质谱法（HPLC/MS）、高压液相色谱/电化学检测法（HPLC/ECD）和气相色谱/质谱法（GC/MS），后两种方法均与固相萃取系统联用，以评估它们对人腰椎脑脊液（CSF）的适用性。此外，对15名对照人员和11名阿尔茨海默病型痴呆（DAT）患者的样本进行了比较，以寻找DAT患者脑脊液中ACh缺乏的情况。GC/MS是最灵敏但最繁琐的方法，而HPLC/ECD的灵敏度可接受、可靠且更具特异性。TS/MS对脑脊液提取物的特异性不足，HPLC/MS更具特异性，但灵敏度远不如前者。因此，只有GC/MS和HPLC/ECD有资格检测人脑脊液提取物中的ACh。GC/MS和HPLC/ECD的比较显示ACh水平高度相关（r = 0.999）。使用HPLC/ECD时，与对照组（6.14±1.39 pmol/ml脑脊液）相比，DAT组的ACh浓度大幅降低（3.75±1.40 pmol/ml脑脊液），但由于低于检测限的样本数量（DAT组11个样本中有8个，对照组15个样本中有7个），对照组和DAT患者之间的差异无统计学意义。两组之间的Ch浓度无统计学意义。数据表明，方法学上的局限性阻碍了在人脑脊液中测定ACh的广泛临床应用。尽管DAT患者脑脊液中的ACh有所降低，但在脑脊液中测定ACh并不适合用于诊断目的。
