Simultaneous analysis of amino acids based on discriminative ¹⁹F NMR spectroscopy

The simultaneous analysis of amino acids (AAs) is crucial for human health, diagnosis and treatment of disease, and nutritional quality evaluation in foodstuffs. Here, we establish an easy and rapid method for the simultaneous analysis of AAs using a single reagent 2-(trifluoromethyl)benzaldehyde (oTFMBA) based on spectral-separation-enabled ¹⁹F NMR spectroscopy. oTFMBA, a highly sensitive chemosensor, is capable of analyzing 19 proteinogenic AAs or non-amino acid amines (non-AAs) in a complex mixture by adjusting the pH in a toilless way. The ¹⁹F signals of oTFMBA-labeled AAs are distributed over a wide range of ∼ 0.7 ppm, demonstrating oTFMBA with higher resolution for simultaneous analysis of AAs compared to the o-phthaldialdehyde (OPA) method (<0.6 ppm). Additionally, 12 AAs were unambiguously identified in human urine, including Asp, Ser, Gly, Thr, Glu, Arg, Ala, Val, Ile, Tyr, His, and Phe. Furthermore, our method's detection limit for AAs is 5.83 μM, illustrating sensitivity with an ∼100-fold improvement over the OPA method. This work represents an approach to the analysis of AAs or non-AAs in a complicated mixture (even biofluid) using a ¹⁹F NMR probe with high sensitivity, which is of great significance for the simultaneous analysis of multiple analytes.