Abstract

In the present study, the magnetic fraction of fly ash was separated from the bulk material using a conventional magnet, yielding approximately 50% w/w. As the magnetic fraction of fly ash contains a high content of aluminum oxide, which is recognized as a phosphate adsorbent, and ferromagnetic iron oxides, it is potentially useful as a magnetic solid-phase extraction adsorbent of phosphoproteins. The results showed that the magnetic fraction of fly ash displayed a high enrichment factor of approximately 130-fold as a magnetic solid-phase extraction adsorbent. The phosphoprotein bound-magnetic fraction was efficiently isolated from the sample solution using a conventional magnet. The adsorbent was further applied to the isolation of phosphoproteins from biological samples, i.e., ovalbumin from egg white and caseins from milk. The sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the enrichment of phosphoproteins revealed that the eluted fraction contained more purified phosphoproteins comparing to the initial diluted samples. In addition, the adsorbent may be reused with acceptable adsorption capacity at least three times. The magnetic solid-phase extraction using the magnetic fraction of fly ash as an adsorbent is effective, simple and reusable. Therefore, the magnetic fraction of fly ash may be an alternative low-cost magnetic adsorbent for the sequestration of phosphoproteins from complex samples.

摘要

在本研究中，使用常规磁体将粉煤灰的磁性部分与大块材料分离，产生约50％w / w。由于粉煤灰的磁性级分包含高含量的氧化铝（被认为是磷酸盐吸附剂）和铁磁性氧化铁，因此潜在地可用作磷蛋白的磁性固相萃取吸附剂。结果表明，作为磁性固相萃取吸附剂，粉煤灰的磁性级分显示出约130倍的高富集系数。使用常规磁体从样品溶液中有效分离出磷蛋白结合的磁性部分。将该吸附剂进一步应用于从生物样品中分离磷蛋白，即从蛋清中分离出卵清蛋白，从牛奶中分离出酪蛋白。磷酸化蛋白的十二烷基硫酸钠-聚丙烯酰胺凝胶电泳显示，与最初的稀释样品相比，洗脱的级分包含更多的纯化的磷酸化蛋白。另外，吸附剂可以以可接受的吸附容量重复使用至少三遍。使用粉煤灰的磁性级分作为吸附剂的磁性固相萃取是有效，简单且可重复使用的。因此，粉煤灰的磁性部分可能是用于从复杂样品中隔离磷蛋白的另一种低成本磁性吸附剂。吸附剂可以以可接受的吸附量重复使用至少三遍。使用粉煤灰的磁性级分作为吸附剂的磁性固相萃取是有效，简单且可重复使用的。因此，粉煤灰的磁性部分可能是用于从复杂样品中隔离磷蛋白的另一种低成本磁性吸附剂。吸附剂可以以可接受的吸附量重复使用至少三遍。使用粉煤灰的磁性级分作为吸附剂的磁性固相萃取是有效，简单且可重复使用的。因此，粉煤灰的磁性部分可能是用于从复杂样品中隔离磷蛋白的另一种低成本磁性吸附剂。