After inserting the sample in the magnet bore, the sample should be allowed to stabilize to the new temperature to allow reproducibility of the results. Next, the magnet should be shimmed properly to ensure a homogeneous field and the probe circuit should be tuned and matched to the transmitter frequency. Improper tuning and matching changes the calibration of the 90° pulses, resulting in variations of the signal intensities. For samples with high ionic strength, tuning and matching may be difficult due to their smaller relative permittivity. This effect is particularly problematic at higher magnetic field strengths. This obstacle may be circumvented by reducing the ionic strength of the sample (e.g. by dilution) or by moving to lower magnetic field strengths. In this paper, all samples were recorded using an 80 MHz benchtop spectrometer to avoid any problems of this kind. Next, the 90° pulse should be calibrated. When choosing signals of protons with different shift for quantification, all signals must have been excited uniformly. Therefore, the excitation profile of the applied pulse should be calculated (from the pulse length and transmitter frequency) and/or verified experimentally. Before the experiment can start, two other very important timings must be set. First, the acquisition time (AQ) must be long enough to ensure that the free induction decay (FID) signal has fully decayed and is not truncated at the end. Truncation of the FID will lead to the presence of so-called sinc-wiggles in the spectrum, affecting the integration area of the signals. Even more important is the other time interval to be set prior to acquisition of the data. This time interval is called the relaxation delay and is no more than a dead time built into the beginning of each pulse sequence. During this time, the nuclei are allowed to relax (back) to their equilibrium position, restoring the z-magnetization. The delay time depends on the longitudinal relaxation time (T1) of the nuclei, an inherent molecular property specific for each nucleus in the sample. To allow for 99% recovery of the equilibrium magnetization, the relaxation delay should be at least five or three times larger than the largest T1 of the sample for a 90° or 30° pulse, respectively. A final acquisition parameter that has to be considered is the number of transients recorded during the experiment. This number should be high enough to allow for an adequate signal-to-noise (SINO) ratio of every analysed signal. As a rule of thumb, we advise the SINO ratio to be larger than 100:1, although this number is instrument specific and should be determined experimentally for every spectrometer setup.