Xevo tq ms system

Fresh, green leaf tissues from each dill genotype were collected in the vegetative stage and freeze-dried at −24 °C (Freeze-dryer Alpha 1–2 LD plus; Christ, Osterode, Germany). Samples were ground to a fine powder using a laboratory grinder mill, IKA A11 (IKA-Werke, Staufen, Germany). The extraction of polyphenolic compounds and LC-MS/MS analysis was accomplished as previously reported Boutsika et al. (2021) (link). Three independent replicates were employed for each dill genotype.
Targeted UPLC analysis was performed on a Waters Acquity UPLC system (Milford, MA, USA) and separation of the phenolic compounds was carried out using a Waters Acquity HSS T3 column (1.8 μm, 100 mm × 2.1 mm), at 40 °C. Phenolic compounds were analyzed as described by Vrhovsek et al. (2012) (link) using water and acetonitrile as mobile phases for the gradient. A Waters Xevo TQMS system equipped with an electrospray (ESI) source was used for mass spectrometry detection. Data was processed using the Mass Lynx Target Lynx Application Manager (Waters).

The method of extracting amino acids from plasma, liver, and muscle was modified from the protocol described by Yuan [26 (link)]. The amino acid analysis was performed using a liquid chromatographic/tandem mass spectroscopic (LC/MS/MS) system for plasma and ultra-performance liquid chromatography (UPLC)/MS system for liver and muscle tissues. The metabolite analysis was performed using an Acquity UPLC System (Waters, Milford, MA, USA) coupled with the Xevo TQ MS system (Waters). For UPLC, a 1.7-mm (2.13100 mm) C18 column (Acquity UPLC System, Waters, Santa Clara, CA, USA) was used. LC separation was carried out at 40 °C at a flow rate of 0.3 mL/min using the following gradient for the analysis: 0–0.5 min 1% B, 0.5–2.5 min from 1% B to 10% B, 2–3.5 min 10% to 35% B, 3.5–6 min from 35% to 99% B, and 6–9 min 1% B (solvent system A: water/formic acid (100:0.1, v/v); B: acetonitrile/formic acid (100:0.1, v/v). Data were acquired with TargetLynx software (Waters, Santa Clara, CA, USA).

The method of extracting amino acids from plasma, liver, and muscle tissues was modified from the protocol of Yuan et al. [55 (link)]. The amino acid analysis was performed by UPLC on an Acquity UPLC System coupled with the Xevo TQ MS system (Waters, Milford, MA, USA). For UPLC, a 1.7 mm (2.13100 mm) C18 column (Waters, Milford, MA, USA) was used. LC separation was carried out at 40 °C with a flow rate of 0.3 mL/min using the following gradient for the analysis: 0~0.5 min 1% B, 0.5~2.5 min from 1% B to 10% B, 2~3.5 min from 10% to 35% B, 3.5~6 min from 35% to 99% B, and 6~9 min 1% B [solvent system A: water/formic acid (100:0.1, vol/vol); B: acetonitrile/formic acid (100:0.1, vol/vol)]. Data were acquired by TargetLynx software (Waters, Milford, MA, USA).