Iconnmr program

Plasma samples were prepared for metabolomic analysis based on nuclear magnetic resonance (NMR) by mixing 3:1 (v/v) of samples and citrate buffer (pH 4.4) using an automated mixer on board. The analysis was performed on an automated high-throughput NMR platform with an Agilent spectrometer 400 MHz (9.4 T), a 4 mm indirect detection probe, and a fixed flow cell (Agilent Technologies, Santa Clara, CA, USA). Data were acquired and processed using Topspin 3.6 (Bruker Biospin), and experiments were performed under automation by the IconNMR program (Bruker Biospin) using the standard pulse sequence, cpmgpr1d (Bruker Biospin).

The spectrometer contained a Bruker AVANCE III HD (Bruker BioSpin GmbH, Rheinstetten, Germany) console combined with a 14.1-T magnet for 1H 600 MHz. It was equipped with a 5-mm inverse triple resonance CryoProbe (1H/13C/15N) with cold preamplifiers for 1H and 13C and a z-axis gradient with automated tuning and matching. The spectrometer contained a Bruker SampleJet system (Bruker Bipspin AG, Fällanden, Switzerland) set to 5-mm shuttle mode with a cooling rack for keeping samples at 279 K. The data were acquired and processed using Topspin 3.2 (Bruker BioSpin GmbH, Rheinstetten, Germany), and experiments were run under automation using the IconNMR program (Bruker BioSpin GmbH, Rheinstetten, Germany). The experiments were conducted using standard pulse sequences, namely noesygppr1d, cpmgpr1d, and zg30 (Topspin 3.2, Bruker Biospin GmbH, Rheinstetten, Germany) [19 (link)].