用于富集磷酸化合物的温度敏感亲和色谱材料的制备及其应用

基于可逆加成断裂链转移聚合(RAFT)法,通过快速引入官能团的方式合成了固定化金属离子温度敏感亲和色谱材料二氧化硅@聚(N-异丙基丙烯酰胺-co-氨甲基膦酸)-钛离子(Ⅳ)(silica@p(NIPAAm-co-AMPA)-Ti⁴⁺),并应用于HPLC中。采用红外光谱、X-射线光电子能谱、热失重和差示量热法对制备的材料进行表征,证明该材料被成功合成并具有较好的温度敏感性能。将该材料用于三磷酸腺苷(ATP)标准品的富集与分离,实现了仅通过改变固定相温度的方式来达到富集与分离小分子磷酸化合物的目的,为富集分离磷酸化肽段奠定了基础。同时该材料还能以在线补充钛离子的方式延长使用寿命。

Preparation and application of temperature-sensitive affinity chromatographic materials for enrichment of phosphoric acid compounds

Based on the surface initiated-reversible addition fragmentation chain transfer (SI-RAFT) method, an immobilized metal ion affinity chromatographic material, silica@ploy(N-isopropylacrylamide-co-(aminomethyl)phosphonic acid)-titanium(Ⅳ) (silica@p(NIPAAm-co-AMPA)-Ti⁴⁺), was synthesized by the method of rapid introduction of functional groups. The synthesis of the material was confirmed by FT-IR and X-ray photoelectron spectroscopy. The grafting ratio and lower critical solution temperature (LCST) of the material were measured by thermal gravimetric analysis (TGA) and differential scanning calorimetry (DSC). The results showed that the material had excellent temperature-sensitive properties and could successfully capture and release adenosine triphosphate successfully. The service life of the material could be extended by online supplementation of Ti⁴⁺.