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1290 infinity 2 uhplc dad

Manufactured by Agilent Technologies
Sourced in Germany

The 1290 Infinity II UHPLC-DAD is an ultra-high-performance liquid chromatography (UHPLC) system with a diode-array detector (DAD) from Agilent Technologies. It is designed to provide high-resolution separation and detection of a wide range of analytes. The system is capable of operating at pressures up to 1,300 bar and flow rates up to 5 mL/min, offering improved separation efficiency and reduced analysis time compared to conventional HPLC systems.

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2 protocols using 1290 infinity 2 uhplc dad

1

Carotenoid Analysis by UHPLC-DAD-MS

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Dried extracts were redissolved in 1 mL MtBE–MeOH (1:1), filtered with a 0.22 μm nylon filter (CellTreat) and 5 μL was injected into an 1290 Infinity II UHPLC-DAD (Agilent; Santa Clara, CA). Carotenoids were separated on a C18 Acquity BEH column (Waters Corp.) 2.1 × 150 mm, 1.7 μm particle size, maintained at 55°C. An isocratic flow using 42% solvent A [80% MeOH, 20% water, and 2% (w/v) aqueous ammonium acetate] and 58% solvent B [78% MtBE, 20% MeOH, and 2% (w/v) aqueous ammonium acetate] at a flow rate of 0.45 mL/min was used, and each run lasted 4.2 min. Quantification was achieved by six-point external calibration curves as described above. Carotenoid identities were confirmed by authentic standards, spectral characteristics, and tandem MS using a 6495 triple quadrupole MS (Agilent) with an atmospheric pressure chemical ionization source operated in positive mode. Source parameters and multiple reaction monitoring experiments were adapted from those previously reported (27 (link)) and were as follows: phytoene: 545.5>463.6, 421.6, 395.6, 327.4; phytofluene: 543.5>461.6, 393.6, 325.4; β-carotene: 537.5>455.3, 269.2, 69.0; and lycopene: 537.5>455.3, 269.2, 69.0.
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2

Quantifying Glucosinolates in Plant Tissues

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To measure glucosinolate compounds, floret and leave samples were grinded with liquid nitrogen and transferred to 2 mL microtubes, lyophilized and stored in a −80 °C freezer. Briefly, 10 mg of lyophilized and homogenized plant material were extracted in presence of 0.02 µmol of the internal standard 4-hydroxybenzyl GS with hot 70% methanol (LC-MS grade, Th. Geyer GmbH & Co. KG, Renningen, Germany) and samples were prepared as described before [36 (link),37 (link)]. The desulfo-GS were analyzed using a 1290 Infinity II UHPLC-DAD coupled with a 6230 ToF-LC/MS (Agilent Technologies, Waldbronn, Germany) with a Poroshell 120 EC-C18 column (Agilent Technologies, Waldbronn, Germany; 100 mm × 2.1 mm, 2.7 μm). UHPLC conditions were as follows: solvent A, MilliQ water; solvent B, 100% v/v acetonitrile. The 19 min run comprised 0.2% (v/v) B (2 min), 0.2% to 19.8% (v/v) B (10 min), a 2 min hold at 19.8% (v/v) B, 19.8% B to 50% (v/v) B (1 min), a 1 min hold at 50% (v/v) B, 50% to 0.2% (v/v) B (1 min), and finally a 2 min hold at 0.2% (v/v) B. The injection volume was 5 µL, and determination was conducted at a flow rate of 0.4 mL min−1 and 30 °C and a wavelength of 229 nm. The concentration of desulfo-GS was calculated by the peak area relative to the area of the internal standard.
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