The analysis of the composition of anthocyanins, phenolic compounds, triterpenic compounds, and phytosterols in cranberry fruit was performed using a Waters ACQUITY Ultra High-Performance LC system (Water, Milford, MA, USA) with a photodiode array detector. The composition of anthocyanins and anthocyanidins in cranberry samples were determined using methodology described by Vilkickyte G. et al. [18 (link)]. Determination of flavonols and chlorogenic acid in cranberry samples was performed by Urbstaitė et al. described methodology [20 (link)]. Triterpenic compounds and phytosterols in cranberry samples was determined using methodology described by Šedbarė et al. [19 (link)].
Acquity ultra high performance lc system
Manufactured by Waters Corporation
Sourced in United States
The ACQUITY Ultra High-Performance LC system is a liquid chromatography instrument designed for high-resolution separation and analysis of complex samples. It utilizes advanced technology to enable fast, efficient, and precise liquid chromatography.
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7 protocols using acquity ultra high performance lc system
1
Comprehensive Phytochemical Analysis of Cranberries
2
Cranberry Phenolic Compound Analysis
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The analysis of the qualitative and quantitative composition of phenolic compounds in cranberry fruit was performed using a Waters ACQUITY Ultra High-Performance LC system (Water, Milford, MA, USA) with a photodiode array detector. An ACE C18 reversed-phase column (ACT, Aberdeen, UK; 100 × 2.1 mm, 1.7 µm particle size) was used for the separation of the compounds at 30 °C. The injection volume was 1 µL, and the distribution was performed using 0.1% formic acid (v/v) (A) and 100% acetonitrile (B) at a flow rate of 0.5 mL/min and the following gradient change: 0 min, 95% A; 1 min, 88% A; 3 min, 88% A; 4 min, 87% A; 9 min, 75% A; 10.5 min, 70% A; 12 min, 70% A; 12.5 min, 10% A; 13 min, 10% A; 13.5 min, 95% A; and 14.5 min, 95% A, delaying the next injection by 2 min.
3
Cranberry Triterpene and Sterol Analysis
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The analysis of the qualitative and quantitative composition of triterpenic compounds, sterols, and squalene in cranberry fruit was performed using a Waters ACQUITY Ultra High-Performance LC system (Water, Milford, MA, USA) with a photodiode array detector and Xevo TQD tandem mass spectrometer (Water, Milford, MA, USA). An Avantor ACE Excel UHPLC column (C18, 100 × 2.1 mm, 1.7 µm particle size) was used for the separation of the compounds at 25 °C. The injection volume was 1 µL, and the distribution was performed using 0.1% formic acid (v/v) (A) and 100% methanol (B) at a flow rate of 0.2 mL/min and the following gradient change: 0 min, 8% A; 8 min, 3% A; 9 min, 2% A; 29.5 min, 2% A; and 30 min, 8% A, delaying the subsequent injection by 10 min. Mass spectrometer conditions were used as follows: a negative electrospray ionization mode with 3 kV capillary voltage and 50 V cone voltage, the source temperature was set to 150° C, the desolvation gas temperature to 600° C, flow to 1000 L/h, and cone gas flow to 20 L/h. MS analysis was performed in negative scan mode in m/z range from 110 to 1000. MS/MS analysis was performed in product ion scan at 40 eV collision energy.
4
Ultra-High-Performance Liquid Chromatography-Tandem Mass Spectrometry Analysis
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The UHPLC analysis was performed via a Waters ACQUITY Ultra High-Performance LC system (Waters, Milford, MA, USA). Separation was achieved on an XBridge® BEH C18 column (3.0 mm × 100 mm, 2.5 µm) at 40 °C with a flow rate of 0.4 mL min−1. The mobile phase consisted of methanol (A) and water containing 5 mmol L−1 ammonium acetate (B). A linear gradient elution program was set as follows: initial 10% A; 1 min, 10% A; 5 min, 90% A; 6 min, 90% A; 6.5 min, 10% A; 8 min, 10% A. The injection volume was 3 µL.
For the MS/MS analysis, a Waters TQS mass spectrometer system (Waters, Milford, MA, USA) was used in positive electrospray ionization mode (ESI+) with the following parameters: interface voltages of capillary, 2.5 kV; source temperature, 150 °C; desolvation temperature, 500 °C. The nebulizing gas and desolvation gas flow rates were 7.0 bar and 1000 L/h, respectively. Multiple reaction monitoring (MRM) mode was used for the quantification and confirmation of the ATs with the parameters shown inTable 3 .
For the MS/MS analysis, a Waters TQS mass spectrometer system (Waters, Milford, MA, USA) was used in positive electrospray ionization mode (ESI+) with the following parameters: interface voltages of capillary, 2.5 kV; source temperature, 150 °C; desolvation temperature, 500 °C. The nebulizing gas and desolvation gas flow rates were 7.0 bar and 1000 L/h, respectively. Multiple reaction monitoring (MRM) mode was used for the quantification and confirmation of the ATs with the parameters shown in
5
UHPLC-MS/MS Analysis of Alternaria Toxins
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UHPLC analysis was performed on a Waters ACQUITY Ultra High-Performance LC system (Waters, Milford, MA, USA). Chromatographic separation was achieved on a Proshell EC18 column (50 mm × 2.1 mm, 2.7 μm). The mobile phase was consisted of methanol (A) and water containing 5 mmol L−1 ammonium acetate (B). A linear gradient elution program was set as follows: initial 10% A; 1 min, 10% A; 5 min, 90% A; 6 min, 90% A; 6.5 min, 10% A; 8 min, 10% A. The flow rate was 0.4 mL min−1. The injection volume was 3 μL, and the column temperature was 35 °C.
For MS/MS detection, a Waters T-QS mass spectrometer system (Waters, Milford, MA, USA) was used both in positive electrospray ionization mode (ESI+) and in negative electrospray ionization mode (ESI−) with the following parameters: interface voltages of capillary, 2.5 kV(ESI+) and 1.5 kV(ESI−); desolvation temperature, 500 °C; and source temperature, 150 °C. The gas flow rates were 7.0 bar for nebulizing gas and 1000 L h−1 for desolvation gas, respectively. Multiple reaction monitoring (MRM) mode was used for the quantification and confirmation of the Alternaria toxins with the parameters shown inTable 1 .
For MS/MS detection, a Waters T-QS mass spectrometer system (Waters, Milford, MA, USA) was used both in positive electrospray ionization mode (ESI+) and in negative electrospray ionization mode (ESI−) with the following parameters: interface voltages of capillary, 2.5 kV(ESI+) and 1.5 kV(ESI−); desolvation temperature, 500 °C; and source temperature, 150 °C. The gas flow rates were 7.0 bar for nebulizing gas and 1000 L h−1 for desolvation gas, respectively. Multiple reaction monitoring (MRM) mode was used for the quantification and confirmation of the Alternaria toxins with the parameters shown in
6
Quantitative Analysis of Fluorophore by LC-MS/MS
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Quantification of FP was carried out by Waters
Xevo TQ tandem quadrupole
mass spectrometer by Waters (Elstree, UK) equipped with an ESI interface,
coupled with a Waters Acquity Ultra High Performance LC system (UPLC),
equipped with a binary solvent delivery system. Chromatographic separations
were carried out on a Waters Acquity UPLC BEH C18 column 130 Å,
1.7 μm, 2.1 × 50 mm. The mobile phase was a mix of mobile
phase A and mobile phase B, which were 0.1% ammonium hydroxide in
water and 1:1 v/v acetonitrile in water, respectively. The flow rate
of the mobile phase was 0.2 mL/min with a 2 min gradient from 50%
to 95% B. Argon was used as the collision gas and the collision energy
was set at 12 V. The LC–MS/MS operations were controlled by
the computer software, MassLynx 4.1, and analyte quantification was
performed with multiple reaction monitoring using the following transitions: m/z 501.4 > 313.1 for FP and m/z 506.4 > 313.1 for FP-d5.
Xevo TQ tandem quadrupole
mass spectrometer by Waters (Elstree, UK) equipped with an ESI interface,
coupled with a Waters Acquity Ultra High Performance LC system (UPLC),
equipped with a binary solvent delivery system. Chromatographic separations
were carried out on a Waters Acquity UPLC BEH C18 column 130 Å,
1.7 μm, 2.1 × 50 mm. The mobile phase was a mix of mobile
phase A and mobile phase B, which were 0.1% ammonium hydroxide in
water and 1:1 v/v acetonitrile in water, respectively. The flow rate
of the mobile phase was 0.2 mL/min with a 2 min gradient from 50%
to 95% B. Argon was used as the collision gas and the collision energy
was set at 12 V. The LC–MS/MS operations were controlled by
the computer software, MassLynx 4.1, and analyte quantification was
performed with multiple reaction monitoring using the following transitions: m/z 501.4 > 313.1 for FP and m/z 506.4 > 313.1 for FP-d5.
7
Quantification of Anthocyanins by UHPLC
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The variability in the qualitative and quantitative content of anthocyanins was evaluated by the validated method [65 (link)]. Chromatographic separation was performed with Waters ACQUITY Ultra-High Performance LC system (Water, Milford, MA, USA) equipped with a photodiode array detector and an ACE Super C18 (100 × 2.1 mm, 1.7 μm) column (ACT, Aberdeen, UK). The gradient elution system consisted of 10% (v/v) formic acid in water (A) and 100% (v/v) acetonitrile (B), and separation was achieved using the following gradient: 0–2 min, 5–9% B; 2–7 min, 9–12% B; 7–9 min, 12–25% B; 9–10 min, 25–80% B; 10–10.5 min, 80% B; 10.5–11 min, 80–5% B; and 11.0–12.0 min, 5% B with flow rate 0.5 mL/min. The column was operated at a constant temperature of 30 °C and the injection volume was 1 μL. All anthocyanins (cyanidin, cyanidin-3-galactoside, cyanidin-3-glucoside, cyanidin-3-arabinoside) were identified and quantified at λ = 520 nm wavelength.
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