Lcms 9030 qtof

The ESI-MS experiments were all performed on a LCMS-9030 qTOF Shimadzu (Shimadzu, Kyoto, Japan) device, equipped with a standard ESI source and the Nexera X2 system. The analysis was performed in the positive ion mode between 50 and 2000 m/z. The LCMS-9030 parameters were as follows: nebulizing gas, nitrogen; nebulizing gas flow, 3.0 L/min; drying gas flow, 10 L/min; heating gas flow, 10 L/min; interface temperature, 300 °C; desolvation line temperature, 400 °C; detector voltage, 2.02 kV; interface voltage, 4.0 kV; collision gas, argon; collision energy was optimized between 10 and 30 eV. The injection volume was 5 μL. The obtained signals all had a mass accuracy error in the range of 1 ppm. The used solvents were all of LC-MS grade. The chromatographic module was operated as follows: eluent (A) water + 0.1% HCOOH, eluent (B) acetonitrile + 0.1% HCOOH. The obtained data were analyzed by LabSolutions software (Shimadzu, Kyoto, Japan).

LC-MS experiments were performed on a LCMS-9030 qTOF Shimadzu (Shimadzu, Kyoto, Japan) device, equipped with a standard ESI source and the Nexera X2 system, equipped with an Aeris Peptide XB-C18 column (50 mm × 2.1 mm) with a 3.6 µm bead diameter equilibrated at 27 °C. The LC system was operated with the following mobile phases, eluent (A) water + 0.1% HCOOH, eluent (B) acetonitrile + 0.1% HCOOH, the gradient conditions (B%) were from 40% to 70% B within 17 min. The flow rate was 0.1 mL/min and the injection volume 0.1 µL. For data analysis, LabSolutions software, version 4.0 was used.

A Shimadzu UHPLC Nexera X2 attached to a Shimadzu (Tokyo, Japan) LCMS-9030 QTOF with ESI source was used. Once samples were filtered (0.45 μm nylon filter, Sigma-Aldrich, St. Louis, MO, USA), betalains were separated using a Shim pack XR–ODS C18 column (150 × 2 mm, 2.2 μm particle size), at 7500 psi maximum working pressure and maximum flow rate of 0.4 mL/min, and using 1% formic acid in water (v/v, eluent A) and a mixture of acetonitrile/water/formic acid (80:19:1) (eluent B) (Sigma-Aldrich, St. Louis, MO, USA). The volume injection was 5 μL. The chromatographic method started with 100% of A, followed by a linear gradient from 0 to 20% of B in 35 min and then a linear gradient from 20 to 100% of B in 5 min [6 (link)]. To re-establish the initial conditions, a linear gradient from 100% of B to 100% of A was used for 10 min. The identification of the individual betalains was performed in positive mode using a sweeping range of m/z 100 to 1000 u. An amount of 4.5 mL/min of nitrogen was used as the drying gas, the temperature was set at 300 °C, the nebulizer gas flow was 3 L/min, the interface temperature was 526 °C, the sampling rate was 5 µ/s, the heater gas flow was 10 L/min, and the detector voltage was 0.20 kV.

As described earlier^{56 (link)}, the statistical analysis of a sub-sample set (102 beers) revealed compounds characteristic for the use of wheat, corn and rice with identification levels reaching from 1 to 3^{109 (link)}. Utilizing the same sample preparation and Shimadzu LCMS-9030 Q ToF (Shimadzu Deutschland GmbH, Duisburg, Germany) analytical system, beers B1885 and B2019 were screened for those marker molecules to verify the carbohydrate source used. For comparison, class-QC samples were analyzed containing all wheat, corn or rice samples, respectively. The measurement parameters are summarized in Supplementary Table S4.

A liquid chromatography-quadrupole time-of-flight tandem mass spectrometer (LC-MS-9030 q-TOF, Shimadzu Corporation, Kyoto, Japan) fitted with a Shim-pack Velox C₁₈ column (100 mm × 2.1 mm with particle size 2.7 μm) was used. The oven temperature was 50 °C. The injection volume was 5 μL and the samples were separated over a 30 min binary solvent gradient. The flow rate was kept constant at 0.3 mL/min using a binary solvent mixture of 0.1 % formic acid in water (Eluent A) and 0.1 % formic acid in methanol (Eluent B). The gradient was gradually increased from 3 to 30 min to facilitate the separation of the compounds within the samples. Briefly, Eluent B was kept at 5 % from 0 to 3 min, gradually increased from 5 to 40 % between 3 and 8 min, and finally increased to 40–95 % between 8 and 23 min. Eluent B was then kept isocratic at 95 % between 23 and 25 min. The gradient was returned to original conditions of 5 % at 25–27 min, and re-equilibration at 5 % occurred at 27–30 min. The liquid chromatographic eluents were then subjected to a Quadruple Time-of-Flight high-definition mass spectrometer for analysis in negative ESI mode. The q-TOF-MS conditions were as follows: 400 °C heat block temperature, 250 °C Desolvation Line (DL) temperature, 42 °C flight tube temperature, and 3 L/min nebulization and dry gas flow.

Electrospray
ionization-mass spectrometry (ESI-MS) experiments were performed on
the LCMS-9030 qTOF Shimadzu (Shimadzu, Kyoto, Japan) device equipped
with a standard ESI source and the Nexera X2 system. Analysis was
performed in the positive ion mode, between 100 and 3000 m/z. LCMS-9030 parameters: nebulizing gas–nitrogen,
nebulizing gas flow 3.0 L/min, drying gas flow—10 L/min, heating
gas flow—10 L/min, interface temperature 300 °C, desolvation
line temperature −400 °C, detector voltage −2.02
kV, interface voltage 4.0 kV, collision gas–argon, mobile phase
(A) H2O +0.1% HCOOH and (B) MeCN +0.1% HCOOH, and mobile phase total
flow −0.3 mL/min. The injection volume was optimized depending
on the intensity of the signals observed on the mass spectrum within
the range of 0.1–1 μL. Obtained signals had a mass accuracy
error in the range of 1 ppm. The concentration of peptide was 0.1
mM, and M/L molar ratio was 1:1. Samples were prepared in a mixture
of water/methanol (50/50 v/v) at pH 7.40. All used solvents were of
LC–MS grade. The obtained data were analyzed by LabSolutions
software (Shimadzu, Kyoto, Japan).

High-resolution mass spectra were obtained on an LCMS-9030 qTOF Shimadzu (Shimadzu, Kyoto, Japan) device, equipped with a standard ESI source and the Nexera X2 system. The mass spectrometer was operated in the positive and negative ion modes. The instrumental parameters were as follows: scan range m/z 100–2000, nebulizing gas nitrogen, nebulizing gas flow 3.0 L/min, drying gas flow 10 L/min, heating gas flow 10 L/min, interface temperature 300 °C, desolvation line temperature 400 °C, detector voltage 2.02 kV, interface voltage 4.0 kV, collision gas argon, mobile phase (A) H₂O + 0.1% HCOOH, (B) MeCN + 0.1% HCOOH, mobile phase total flow 0.3 mL/min. The injection volume was optimized depending on the intensity of the signals observed on the mass spectrum within the range of 0.1 to 3 μL. The samples were prepared in a 1:1 methanol-water mixture at a pH value of 7.4. The sample concentration was [ligand]_tot = 0.1 M, and M:L molar ratio was 1:1. Data were processed using the ACDLabs Spectrus Processor v2021.1.3 program. A comparison between the obtained experimental signals and the true isotopic pattern calculated using Bruker Compass DataAnalysis 3.4 program enabled an unambiguous confirmation of the elemental composition of the obtained complex (Table S2, Figures S15–S17, Supplementary Materials).