Qualitative determination of early Maillard-products by MALDI-TOF mass spectrometry peptide mapping
Qualitative determination of early Maillard-products by MALDI-TOF mass spectrometry peptide mapping
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DOI：
10.1007/s00217-002-0491-4
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发表时间：
2002-07-01
影响因子：
3.3
通讯作者：
Pischetsrieder, M
中科院分区：
文献类型：
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作者：
Kislinger, T;Humeny, A;Pischetsrieder, M
采用基质辅助激光解吸电离飞行时间质谱（MALDI-TOF MS）定性研究了溶菌酶与7种不同糖（D-核糖、L-鼠李糖、D-葡萄糖、D-果糖、D-半乳糖、D-乳糖、D-麦芽糖）在有氧条件下孵育后早期美拉德产物的形成。完整溶菌酶的MALDI-TOF质谱显示了糖基化产物的不均匀分布，检测到显着的质量增加和峰增宽。因此，为了获得关于糖基化产物性质的更详细信息，我们进行了MALDI-TOF MS肽图谱分析。在每个肽图谱中，我们能够以高选择性检测早期糖化产物（Amadori产物/葡糖胺）。由于糖化肽的特征性质量增加，我们能够区分不同糖的Amadori产品。此外，观察到修饰模式的变化，很可能是由于七种还原糖的不同相对性。还检测到最初形成的美拉德产物的其他更高级的糖基化产物。关于这些晚期美拉德产物的性质的进一步研究正在进行中。在这里，我们报告，第一次，使用MALDI-TOF MS肽图谱作为一种快速和高度选择性的方法检测早期美拉德产品的溶菌酶与7种不同的还原糖孵育后产生的。
Matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF MS) was used to qualitatively study the formation of early Maillard products of lysozyme, produced upon incubation with seven different sugars (D-ribose, L-rhamnose, D-glucose, D-fructose, D-galactose, D-lactose, D-maltose) in solution, in the presence of oxygen. MALDI-TOF mass spectra of intact lysozyme revealed a heterogeneous distribution of glycation products, detected by significant mass increase and peak broadening. Therefore, to get more detailed information about the nature of the glycation products, we performed MALDI-TOF MS peptide mapping analysis. In each of the peptide mapping spectra we were able to detect the early glycation products (Amadori products/glucosylamines) with high selectivity. We were able to distinguish between the Amadori products of the different sugars, due to the characteristic mass increases of the glycated peptides. Furthermore, variations of the modification pattern were observed, very likely due to the different relativities of the seven reducing sugars. Other, more advanced glycation products of the initially formed Maillard products were also detected. Further studies regarding the nature of these late Maillard products are in progress. Here we report, for the first time, the use of MALDI-TOF MS peptide mapping as a quick and highly selective method for the detection of early stage Maillard products produced upon incubation of lysozyme with seven different reducing sugars.