Samplexpress

All the solvents and reagents are of high purity and have been purchased commercially from Sigma-Aldrich, Fluorochem, Alfa Aesar and Fisher Scientific. Melting points were recorded using a Gallen kamp melting point apparatus and are uncorrected. ¹H and ¹³C NMR analyses were performed on a 400 MHz spectrometer (Bruker) equipped with a SampleXpress (from Bruker) auto sampler system, using deuterated solvents for the preparation of the samples. The obtained spectra were analyzed using Topspin 7.1 software (Bruker). The chemical shifts were reported in ppm values relative to tetramethylsilane (TMS), used as internal standard. Signals were identified and described as singlet (s), doublet (d), triplet (t) and multiplet (m). Coupling constants were reported in Hertz (Hz). Mass Spectra (ESI), HRMS Agilent technologies 6890N and an inert mass selective detector 5973 mass spectrometer Technologies. Glassware used was dried in a UN55 oven (Memmert) at 200 °C^{35 (link)}. Fourier transform Infrared (FTIR) spectrum of the synthesized compounds was taken using KBr pellet press method by Bruker TENSOR 27 FTIR spectrophotometer and the (%) yields are calculated on the basis of 1.0 mM of each reactant used. The synthesized motifs 3(a–u) were characterized as given below:

The proton NMR (¹H NMR) data acquisition was carried out using a JEOL ECS 400 MHz NMR spectrometer at 25°C with a sensitivity of 280 (0.1% ethyl benzene) for ¹H and ¹⁹F, with an automatic Bruker SampleXpress sample charger run by using a 500 MHz electric DC motor having a 60 sample carousel controlled using ICON-NMR software and equipped with barcode reader registration, with the samples being kept at a temperature between 5 and 30°C and a separate cryo-fit mounting kit for sample cooling. The sample charger and sample unit were both controlled by the Bruker Avance III 400 MHz controller unit.
The sample (∼5 mg) was dissolved in 1-cm³ wet CDCl₃ under atmospheric conditions; no special precautions were taken other than that, and the sample was initially transferred into the NMR tube in an Ar glove box.

All plasma and urine samples were analyzed by 1D-¹H-NMR spectroscopy. Plasma samples were measured at 310 K on an AVANCE II 600 MHz NMR spectrometer equipped with a ¹H-BBI probehead and a BACS sample changer (Bruker BioSpin GmbH, Rheinstetten, Germany). Urine samples were analyzed at 300 K on a Bruker 600 MHz spectrometer (either AVANCE III equipped with a ¹H,¹³C,¹⁵N-TCI inversely detected cryoprobe or AVANCE II with ¹H-BBI room temperature probe (Bruker BioSpin GmbH, Rheinstetten, Germany)) equipped with either SampleXpress or BACS sample changer, respectively, as described in Rist et al. [25 (link)]. Typically, metabolites that can be detected include organic acids, amino acids, amines, sugars, sugar alcohols, and others. Method details can be found in section H of the S1 File.