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Application of matrix solid-phase dispersion methodology to the extraction of endogenous peptides from porcine hypothalamus samples for MS and LC-MS analysis

机译:基质固相分散法在猪下丘脑样品中提取内源性肽用于MS和LC-MS分析的应用

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摘要

In this study, we investigated a novel application of matrix solid-phase dispersion (MSPD) methodology for the extraction of endogenous peptides from porcine hypothalamus tissue samples. Several experimental factors of the MSPD procedure were examined. Finally, silica-based octadecyl was chosen as dispersing material and blended with 0.25. g porcine hypothalamus at a ratio of 5, and 10. mL of 60% acetonitrile with 0.2% formic acid in water was chosen as the extraction and elution solvent. This MSPD extraction method was compared to the classic acid extraction method. More peaks were observed in the MSPD extracts (74 ± 5) by MALDI-TOF MS than in acid extracts (34 ± 5). Moreover, 14 potential endogenous peptides were identified in the MSPD extracts after nanoLC-MS/MS analysis, while only 2 endogenous peptides in the acid extracts. These results indicated that MSPD could be employed as a simple and efficient method for the extraction of endogenous peptides from tissues.
机译:在这项研究中,我们调查了基质固相分散(MSPD)方法在猪下丘脑组织样品中提取内源肽的新应用。检查了MSPD程序的几个实验因素。最后,选择基于二氧化硅的十八烷基作为分散材料并与0.25混合。取猪下丘脑(比例为5)和10。10mL 60%乙腈和0.2%甲酸的水溶液作为提取和洗脱溶剂。将该MSPD提取方法与经典酸提取方法进行了比较。通过MALDI-TOF MS在MSPD提取物中(74±5)观察到的峰比在酸性提取物中(34±5)观察到的更多。此外,经过nanoLC-MS / MS分析后,在MSPD提取物中鉴定出14种潜在的内源肽,而在酸性提取物中仅鉴定出2种潜在内源肽。这些结果表明,MSPD可作为一种简单有效的方法从组织中提取内源性肽。

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