State Key Laboratory of Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University, Changsha 410082, PR China.
State Key Laboratory of Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University, Changsha 410082, PR China.
Talanta. 2018 May 15;182:484-491. doi: 10.1016/j.talanta.2018.02.022. Epub 2018 Feb 8.
A novel magnetic solid-phase extraction (MSPE) method based on 1-hexyl-3-methyl imidazolium chloride ionic liquid (IL) modified magnetic FeO nanoparticles, hydroxylated multiwall carbon nanotubes (MWCNTs-OH) and zeolitic imidazolate frameworks (ZIFs) nanocomposites (FeO-MWCNTs-OH@ZIF-67@IL) were proposed and applied to extract α-chymotrypsin. The magnetic materials were synthesized successfully and characterized by X-ray diffraction (XRD), transmission electron microscope (TEM), thermal gravimetric analysis (TGA), fourier transform infrared spectrometry (FT-IR), vibrating sample magnetometer (VSM) and zeta potentials. Subsequently, the UV-vis spectrophotometer at about 280 nm was utilized to quantitatively analyze the α-chymotrypsin concentration in the supernatant. Furthermore, single factor experiments revealed that the extraction capacity was influenced by initial α-chymotrypsin concentration, ionic strength, extraction time, extraction temperature and pH value. The extraction capacity could reach up to about 635 mg g under the optimized conditions, absolutely higher than that of extraction for Ovalbumin (OVA), Bovine serum albumin (BSA) and Bovine hemoglobin (BHb). In addition, the regeneration studies showed FeO-MWCNTs-OH@ZIF-67@IL particles could be reused several times and kept a high extraction capacity. Besides, the study of enzymatic activity also indicated that the activity of the extracted α-chymotrypsin was well maintained 93% of initial activity. What's more, the proposed method was successfully applied to extract α-chymotrypsin in porcine pancreas crude extract with satisfactory results. All of above conclusions highlight the great potential of the proposed FeO-MWCNTs-OH@ZIF-67@IL-MSPE method in the analysis of biomolecules.
一种基于 1-己基-3-甲基氯化咪唑离子液体（IL）修饰的磁性 FeO 纳米粒子、羟基化多壁碳纳米管（MWCNTs-OH）和沸石咪唑酯骨架（ZIFs）纳米复合材料（FeO-MWCNTs-OH@ZIF-67@IL）的新型磁性固相萃取（MSPE）方法被提出并应用于提取α-糜蛋白酶。磁性材料通过 X 射线衍射（XRD）、透射电子显微镜（TEM）、热重分析（TGA）、傅里叶变换红外光谱（FT-IR）、振动样品磁强计（VSM）和zeta 电位进行了成功的合成和表征。随后，在大约 280nm 处利用紫外-可见分光光度计定量分析上清液中α-糜蛋白酶的浓度。此外，单因素实验表明，萃取能力受初始α-糜蛋白酶浓度、离子强度、萃取时间、萃取温度和 pH 值的影响。在优化条件下，萃取容量可达约 635mg/g，绝对高于对卵清蛋白（OVA）、牛血清白蛋白（BSA）和牛血红蛋白（BHb）的萃取。此外，再生研究表明，FeO-MWCNTs-OH@ZIF-67@IL 颗粒可以重复使用几次，并保持较高的萃取能力。此外，酶活性研究也表明，提取的α-糜蛋白酶的活性保持在初始活性的 93%左右。更重要的是，该方法成功地应用于猪胰腺粗提物中α-糜蛋白酶的提取，结果令人满意。所有这些结论都凸显了所提出的 FeO-MWCNTs-OH@ZIF-67@IL-MSPE 方法在生物分子分析中的巨大潜力。
