Schiller J, Hammerschmidt S, Wirtz H, Arnhold J, Arnold K
Institute of Medical Physics and Biophysics, Medical Department, University of Leipzig, Liebigstr. 27, D-04103 Leipzig, Germany.
Chem Phys Lipids. 2001 Jul;112(1):67-79. doi: 10.1016/s0009-3084(01)00163-3.
Despite the high clinical relevance, only the cellular moiety of bronchoalveolar lavage (BAL) has been intensively investigated and is used for diagnosis purposes. On the other hand, the cell-free fluid is, by far, less characterized. Although this fluid represents a relatively simple mixture of only a few different phospholipids (mainly phosphatidylcholine, phosphatidylglycerol and cholesterol), methods for the routine analysis of these fluids are still lacking. In the present investigation we have applied, for the first time, MALDI-TOF mass spectrometry, as well as 31P NMR spectroscopy to the analysis of organic extracts of bronchoalveolar lavage fluids. BAL from different mammals (rat, minipig, rabbit and man) were investigated and, for means of comparison, organic extracts of lung tissue were also examined. Both applied methods provide fast and reliable information on the lipid composition of the bronchoalveolar lavage. However, despite of its comparably low sensitivity, 31P NMR spectroscopy detects all phospholipid species in a single experiment and with the same sensitivity, whereas MALDI-TOF fails in the detection of phosphatidylethanolamine in the presence of higher quantities of phosphatidylcholine. In contrast, MALDI-TOF mass spectrometry is more suitable for the detection of cholesterol and the determination of the fatty acid composition of the individual phospholipids, especially lysolipids. It will be shown that all BALs exhibit significant, species-dependent differences that mainly concern the content of phosphatidylglycerol and lyso-phosphatidylcholine. It is concluded that both methods are suitable tools in lipid research due to the (in comparison to alternative methods) simplicity of performance.
尽管具有很高的临床相关性,但目前仅对支气管肺泡灌洗(BAL)的细胞部分进行了深入研究并用于诊断目的。另一方面,迄今为止,对无细胞液的特征了解较少。虽然这种液体仅由几种不同的磷脂(主要是磷脂酰胆碱、磷脂酰甘油和胆固醇)组成,是一种相对简单的混合物,但仍缺乏对这些液体进行常规分析的方法。在本研究中,我们首次将基质辅助激光解吸电离飞行时间质谱(MALDI-TOF)以及31P核磁共振光谱(31P NMR)应用于支气管肺泡灌洗液体有机提取物的分析。研究了来自不同哺乳动物(大鼠、小型猪、兔子和人类)的BAL,并且为了进行比较,还检测了肺组织的有机提取物。两种应用方法都能快速可靠地提供有关支气管肺泡灌洗脂质组成的信息。然而,尽管31P NMR光谱的灵敏度相对较低,但它能在一次实验中以相同的灵敏度检测所有磷脂种类,而在磷脂酰胆碱含量较高时,MALDI-TOF无法检测到磷脂酰乙醇胺。相比之下,MALDI-TOF质谱更适合检测胆固醇以及确定单个磷脂(尤其是溶血磷脂)的脂肪酸组成。结果表明,所有的BAL都表现出显著的、物种依赖性差异,主要涉及磷脂酰甘油和溶血磷脂酰胆碱的含量。得出的结论是,由于(与其他方法相比)操作简单,这两种方法都是脂质研究的合适工具。