利用金屬有機骨架進行分散式固相萃取搭配液相層析串聯式質譜技術分析單、雙及三磷酸腺苷化合物

Abstract

核苷酸化合物是生物系統中作為能量來源的基本生物分子，由核糖、含氮鹼基及磷酸基團組成，存在於生物體如植物、動物、細菌當中。金屬有機骨架（MOFs）由於其高表面積和均勻的孔徑，在吸附領域當中有廣泛的應用。近年来的文獻顯示出MIL-101-NH2 (Al) 從水中去除磷酸鹽的顯著潛力，而腺苷核苷酸化合物結構上帶有磷酸基團。本研究中，合成MIL-101-NH2 (Al)及MIL-160 (Al)並對其進行表徵。同時選擇 MIL-101-NH2 (Al)作為分散式固相萃取(Dispersive Solid-Phase Extraction, dSPE)的吸附劑，以 MIL-160 (Al)作為對照。使用cyanopropyl(CN)管柱並搭配超高效液相層析串聯質譜(UPLC-MS/MS)系統分對AMP、ADP 和 ATP三個核苷酸化合物進行分析。MOF搭配分散式固相萃取法進行優化結果表明，結果顯示在酸性環境下AMP、ADP 和 ATP 能夠被成功脫附出來。同時層析上能在14分鐘內實現了良好的分離，並且具有良好的準確性(82-121%)及精密度(0.4%-14.3%)，線性範圍為 1-1000 ng/mL，相關係數r> 0.995。偵測極限和定量極限分別為 0.5 ng/mL 和 1 ng/mL。本研究開發的基於 MOF 的 dSPE 搭配UPLC-MS/MS 方法在食品或生物樣品中具有巨大的應用潛力。

Nucleotides are essential biomolecules that serve as sources of energy in biological systems. Metal-organic frameworks (MOFs), have garnered significant attention due to their high surface area and uniform pore size. The MOFs is an ideal material for adsorp-tion applications, which have demonstrated significant potential of MIL-101-NH2 (Al) for phosphate removal from water. In this study, MIL-101-NH2 (Al) and MIL-160 (AI) were synthesized and characterized. MIL-101-NH2 (Al) selected as dispersive solid-phase extraction (dSPE) adsorbent, MIL-160 (AI) was used for comparison. The MOF-based dSPE method exhibited excellent efficiency in eluting nucleotides under acidic conditions. Ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) system was used for the determination of AMP, ADP and ATP. A good separation was achieved within 14 minutes, demonstrating high accuracy (82%-121%) and precision (0.4%-14.3%), with a linear range of 1-1000 ng/mL and correla-tion coefficients (r)> 0.995. The limit of detection (LOD) and limit of quantification (LOQ) were determined to be 0.5 ng/mL and 1 ng/mL, respectively. The MOF-based dSPE and UPLC-MS/MS method developed in this study holds immense potential for further applications in food or biological samples.