Musharraf Syed Ghulam, Bibi Aisha, Shahid Najia, Najam-Ul-Haq Muhammad, Ambreen Nida, Khan Momin, Khan Khalid Mohammed, Choudhary M Iqbal, Rahman Atta Ur
Dr, Panjwani Center for Molecular Medicine and Drug Research, International Center for Chemical and Biological Sciences, University of Karachi, Karachi 75270, Pakistan.
Chem Cent J. 2013 Apr 26;7(1):77. doi: 10.1186/1752-153X-7-77.
Matrix-assisted laser desorption/ionization (MALDI) is a soft ionization mass spectrometric technique, allowing the analysis of bio-molecules and other macromolecules. The matrix molecules require certain characteristic features to serve in the laser desorption/ionization mechanism. Therefore, only a limited number of compounds have been identified as ultraviolet- laser desorption/ionization (UV-LDI) matrices. However, many of these routine matrices generate background signals that useful information is often lost in them. We have reported flavones, coumarindione and benzimidazole derivatives as alternate UV-LDI matrices.
Thirty one compounds have been successfully employed by us as matrices for the analysis of low molecular weight (LMW) peptides (up to 2000 Da). Two peptides, bradykinin and renin substrate tetra-decapeptide were analyzed by using the newly developed matrices. The MS measurements were made after mixing the matrix solution with analyte by using dried droplet sample preparation procedures. The synthesized matrix materials showed better S/N ratios and minimal background signals for low mass range. Furthermore, pico molar concentrations of [Glu1]-fibrinopeptide B human could be easily analyzed with these matrices. Finally, BSA-digest was analyzed and identified through database search against Swiss-Prot by using Mascot.
These results validate the good performance of the synthesized UV-laser desorption/ionization (LDI) matrices for the analysis of low molecular weight peptides.
基质辅助激光解吸/电离(MALDI)是一种软电离质谱技术,可用于分析生物分子和其他大分子。基质分子需要具备某些特征才能参与激光解吸/电离机制。因此,只有有限数量的化合物被鉴定为紫外激光解吸/电离(UV-LDI)基质。然而,许多这些常规基质会产生背景信号,有用信息常常在其中丢失。我们已报道黄酮类、香豆素二酮和苯并咪唑衍生物可作为替代的UV-LDI基质。
我们已成功将31种化合物用作分析低分子量(LMW)肽(分子量高达2000 Da)的基质。使用新开发的基质对两种肽,即缓激肽和肾素底物十四肽进行了分析。通过使用干滴样品制备程序将基质溶液与分析物混合后进行质谱测量。合成的基质材料在低质量范围内显示出更好的信噪比和最小的背景信号。此外,使用这些基质可以轻松分析皮摩尔浓度的人[Glu1]-纤维蛋白肽B。最后,通过使用Mascot针对Swiss-Prot数据库搜索对牛血清白蛋白消化产物进行了分析和鉴定。
这些结果验证了合成的紫外激光解吸/电离(LDI)基质在分析低分子量肽方面的良好性能。