Stahl B, Linos A, Karas M, Hillenkamp F, Steup M
Institute for Medical Physics and Biophysics, University of Muenster, Germany.
Anal Biochem. 1997 Mar 15;246(2):195-204. doi: 10.1006/abio.1997.2011.
In this communication both matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) and high-performance anion-exchange chromatography (HPAEC) have been applied to analyze fructans from higher plants. Size distribution of a commercially available fructan preparation from Dahlia variabilis L. was determined by MALDI-MS. Molecular masses ranged from 2,000 up to 6,000 Da with a peak value of distribution at 2,635 Da. Essentially the same pattern was obtained using HPAEC. Low-molecular-weight fructans from onion bulbs (Allium cepa L.) were studied in more detail. Tissue extracts were analyzed by MALDI-MS without any analyte purification. Mass-spectra of both proteins and oligosaccharides were obtained. For identification, metastable ion scanning was performed. Neither deproteinization nor deionization of the samples affected the oligosaccharide pattern. Using HPAEC, a more complex oligosaccharide pattern was obtained because isomeric glycans were differentiated. However, the overall size distribution was similar to that obtained by MALDI-MS. In further experiments epidermal or parenchyma cell layers of the onion bulb were placed into matrix solution and were then subjected to MALDI-MS and metastable ion scanning as well. By taking this approach, analyte desorption was achieved immediately from plant tissue. Oligosaccharide mass spectra were essentially the same as those of the extracts. To our knowledge, this is the first time that MALDI-MS has been applied as a microprobe to plant tissue. Finally MALDI-MS analysis was performed using single-cell extracts from onion tissues without any purification of the analyte.
在本通讯中,基质辅助激光解吸/电离质谱法(MALDI-MS)和高效阴离子交换色谱法(HPAEC)都已用于分析高等植物中的果聚糖。通过MALDI-MS测定了一种市售的大丽花属植物果聚糖制剂的大小分布。分子量范围为2000至6000 Da,分布峰值为2635 Da。使用HPAEC获得了基本相同的图谱。对洋葱鳞茎(葱属植物)中的低分子量果聚糖进行了更详细的研究。在不进行任何分析物纯化的情况下,通过MALDI-MS对组织提取物进行分析。获得了蛋白质和寡糖的质谱图。为了进行鉴定,进行了亚稳离子扫描。样品的脱蛋白和去离子处理均未影响寡糖图谱。使用HPAEC,由于异构聚糖得以区分,因此获得了更复杂的寡糖图谱。然而,总体大小分布与通过MALDI-MS获得的相似。在进一步的实验中,将洋葱鳞茎的表皮或薄壁细胞层置于基质溶液中,然后也进行MALDI-MS和亚稳离子扫描。通过这种方法,可立即从植物组织中实现分析物解吸。寡糖质谱图与提取物的基本相同。据我们所知,这是首次将MALDI-MS用作植物组织的微探针。最后,使用未经任何分析物纯化的洋葱组织单细胞提取物进行MALDI-MS分析。