Enzyme assays are important for studying enzymemediated biochemical reactions and for clinical diagnosis and drug development. The technique of an immobilized enzyme microreactor (IMER) integrated with capillary electrophoresis (CE) has been frequently utilized in online enzyme assays. However, the traditional approaches for IMER-CE enzyme analysis have some defects such as low loading capacity and poor stability. Herein, metal−organic frameworks (MOFs), which have enormous potential in the fields of enzyme immobilization and capillary electrochromatographic (CEC) separation, were first explored as novel support materials with good enzyme immobilization performance and stationary phases with excellent separation abilities to construct an integrated MOFs-IMER-CEC microanalysis system for a high-efficiency online enzyme assay. As a proof-of-concept demonstration, acetylcholinesterase (AChE) was immobilized on a densely packed UiO-66-NH 2 nanocrystal coating on a capillary inner surface with abundant intercrystalline mesoporosity and was employed as a highly effective and robust IMER for CEC-integrated online enzyme analysis. The excellent separation performance of the UiO-66-NH 2 -modified capillary was verified by high-efficiency separation of three types of neutral, acidic, and basic compounds. The Michaelis−Menten constant and enzyme inhibition kinetics of UiO-66-NH 2 −IMER were systematically assessed, exhibiting distinct advantages such as remarkably increased enzyme loadability, superior affinity for substrates, and greatly improved stability and repeatability compared to CE-integrated IMERs prepared by the traditional covalent bonding method. Furthermore, the developed method was successfully utilized for detecting organophosphorus pesticides in leguminous vegetable samples, demonstrating its strong practicality. The study not only proposed a novel support material and construction strategy for a highperformance microchannel-based IMER but also can be widely used in bioanalysis and biosensing research.
A large
number of metal–organic frameworks (MOFs) have exhibited
increasingly wide utilization in the field of chromatographic separation
owing to their intrinsic fascinating properties. However, the previous
studies on supported MOF coating-based chromatographic separation
focused only on the synthesis and chromatographic performance of a
certain kind of supported MOF coatings as stationary phases using
the multiple-step, complicated, and time-consuming modification methods,
which severely impeded the widespread application of MOFs in separation
science. Herein, a high-efficiency and versatile methodology toward
diverse supported MOF coating-based stationary phases to achieve high-efficiency
chromatographic separation was first reported based on the immobilized
cysteine (Cys)-triggered in situ growth (ICISG) strategy. As a proof-of-concept
demonstration, four types of MOF crystals consisting of different
ligands and metal ions (Zn2+, Cu2+, Fe3+, and Zr4+) were conveniently and firmly grown on a Cys-modified
capillary using the ICISG strategy and employed as the functional
stationary phase for electrochromatographic separation. A broad variety
of neutral, acidic, and basic compounds were all separated in a highly
efficient manner on the developed four MOF-coated columns. The maximum
theoretical plate number for Cys-MIL-100(Fe)@capillary was close to
1.0 × 105 plates/m, and the intraday, interday, and
column-to-column repeatabilities of retention times for the four MOF-modified
columns were all less than 5.25%. More interestingly, the diversified
separation performance of the developed MOF-coated columns indicated
that the preparation strategy and the skeletal structure of the MOF
coating-based stationary phases have a significant influence on the
electrochromatographic separation performance and column capacity.
Benefiting from the strong universality and high applicability of
the developed ICISG strategy, the present study provides an effective
route to facilitate the design and fabrication of novel functional
MOF-based chromatographic stationary phases.
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