The HPLC-MS analyses were carried out using an Agilent 1100 series LC-MSD trap VL, (Agilent Technologies, Santa Clara, CA, USA), equipped with a G1322A degasser, a G1311A QuatPump, a G1313A ALS, a G1316A Colcom, a G1315A DAD and a Kromasil 100–5 C18 column (250 × 4.6 mm, 5 μm). The mobile phases were: (A) formic acid/acetonitrile/water (2:6:92, v/v/v), and (B) formic acid/acetonitrile/water (2:54:44, v/v/v). The injection volume was 30 μL with a flow rate of 1.0 mL/min. The gradient elution of solvent B was applied as follows: 1–18 min, 10% to 25%; 18–20 min, 25%; 20–30 min, 25% to 40%; 30–35 min, 40% to 70% and 35–40 min, 70% to 100%. The column temperature was 50 °C and the detection wavelength was 525 nm. MS conditions were as follows: Electrospray ionization (ESI) interface, positive ion mode, 35 psi nebulizer pressure, 10 mL/min dry gas flow rate, 300 °C dry gas temperature, and scans at m/z 100–1000 [28 (link),32 (link),43 (link)].
Agilent 1100 series lc msd trap vl
Manufactured by Agilent Technologies
Sourced in United States
The Agilent 1100 series LC-MSD trap VL is a liquid chromatography-mass spectrometry (LC-MS) system designed for analytical applications. It features a variable-loop ion trap mass spectrometer that provides high-performance mass analysis capabilities. The system is equipped with an integrated liquid chromatography module for sample separation and introduction into the mass spectrometer.
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4 protocols using agilent 1100 series lc msd trap vl
1
HPLC-MS Analysis of Compounds
2
Grape Anthocyanin Profiling Across Maturation
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Grapes at 8, 12, and 16 WAA of 5 cultivars were used to determine individual anthocyanins. Specific extraction steps have been described previously [20 (link)]. Three independent samples were extracted for each skin, and were stored at −40 °C until analysis. Before injection, the extracts were filtered through 0.45 µm filters (cellulose acetate and nitrocellulose, CAN).
Individual anthocyanins of extracts were analyzed using an Agilent 1100 series LC-MSD trap VL equipped with a diode array detector and a Kromasil C18 column (250 × 4.6 mm, 5 μm) (Agilent Technologies, Santa Clara, CA, USA) utilizing a binary solvent gradient, where mobile phase A was 2% formic acid in water and B was 2% formic acid in acetonitrile. The detailed LC procedures and MS conditions have been described previously report [20 (link)].
The elution order and retention time of the anthocyanins were compared with those of the standard, and the weights of molecular ions and fragment ions were compared with the weights of standard and reference products, allowing different anthocyanins to be identified [25 (link),26 (link)]. Anthocyanins were quantified using malvidin-3-O glucoside as an external standard.
Individual anthocyanins of extracts were analyzed using an Agilent 1100 series LC-MSD trap VL equipped with a diode array detector and a Kromasil C18 column (250 × 4.6 mm, 5 μm) (Agilent Technologies, Santa Clara, CA, USA) utilizing a binary solvent gradient, where mobile phase A was 2% formic acid in water and B was 2% formic acid in acetonitrile. The detailed LC procedures and MS conditions have been described previously report [20 (link)].
The elution order and retention time of the anthocyanins were compared with those of the standard, and the weights of molecular ions and fragment ions were compared with the weights of standard and reference products, allowing different anthocyanins to be identified [25 (link),26 (link)]. Anthocyanins were quantified using malvidin-3-O glucoside as an external standard.
3
Quantification of Polyphenol Classes
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Anthocyanins and flavonols were analyzed on an Agilent 1100 series LC-MSD trap VL equipped with a diode array detector and a Kromasil C 18 column (250 × 4.6 mm, 5 μm) (Agilent Technologies, Santa Clara, CA, USA) utilizing a binary solvent gradient, where mobile phase A was 2% formic acid in water and B was 2% formic acid in acetonitrile. The detailed LC procedures and MS conditions have been described previously (He et al., 2010) (link). The flavan-3-ols were tested using the Agilent 1200 Series LC-MSD trap VL equipped with a 6410 Triple Quadrupole Mass Spectrometer (QqQ). The column used was an Agilent Poroshell 120 EC-C 18 column (150 × 3.0 mm, 2.7 μm). The detailed LC procedures and MS conditions have been described previously (Li et al., 2016) (link). Flavonols and anthocyanins were quantified using quercetin-3-O-glucoside and malvidin-3-Oglucoside as external standards, respectively. The content of flavan-3-ols was quantified using catechin (C), epicatechin (EC), epigallocatechin (EGC), and epicatechin-3-O-galate (ECG) as external standards.
4
Anthocyanin Extraction and HPLC-MS Analysis
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The extraction of anthocyanins in grape skins and the HPLC-MS analysis were carried out according to the previously published method of He et al. (2010) (link). An Agilent 1100 series LC-MSD trap VL (Agilent, Santa Clara, CA, USA), equipped with a G1379A degasser, a G1312BA QuatPump, a G1313A ALS, a G1316A column, a G1315A DAD and a Kromasil 100-5 C18 column (250 × 4.6 mm, 5 μm) was used for anthocyanins detection. A flow rate of l mL/min at ambient temperature was used. Solvent A was 2% (v:v) formic and 6% acetonitrile in water, and solvent B was acetonitrile containing 2% formic acid and 44% water. Proportions of solvent B varied as follows:
1-18 min, 10% to 25%; 18-20 min, 25%; 20-30 min, 25% to 40%; 30-35 min, 40% to 70% and 35-40 min, 70% to 100%. Onjection volumes were 30 μL, and the detection wavelength was 525 nm. The column temperature was 50 °C. MS conditions were as follows: Electrospray ionisation (ESO) interface, positive ion model, 30 psi nebulizer pressure, 12 mL/min dry gas flow rate, 300 °C dry gas temperature, and scans at m/z 100-1500. Anthocyanins were quantified at 525 nm as malvidin-3-D-glucoside using calibration curves obtained within a concentration range between 0.5 and 1000 mg/L, with linear correlation coefficients greater than 0.997. Both standards and samples were determined in triplicate.
1-18 min, 10% to 25%; 18-20 min, 25%; 20-30 min, 25% to 40%; 30-35 min, 40% to 70% and 35-40 min, 70% to 100%. Onjection volumes were 30 μL, and the detection wavelength was 525 nm. The column temperature was 50 °C. MS conditions were as follows: Electrospray ionisation (ESO) interface, positive ion model, 30 psi nebulizer pressure, 12 mL/min dry gas flow rate, 300 °C dry gas temperature, and scans at m/z 100-1500. Anthocyanins were quantified at 525 nm as malvidin-3-D-glucoside using calibration curves obtained within a concentration range between 0.5 and 1000 mg/L, with linear correlation coefficients greater than 0.997. Both standards and samples were determined in triplicate.
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