Fifty-microliter-aliquots of recombinant arylsulfatases in 100 mM Tris, pH 7.5, were mixed with 50 µl of 5 mM GS solution or 50 µl of GS extracts for 2 h at ambient temperature. Reactions were stopped with 500 µl methanol and mixtures were centrifuged for 5 min. Two hundred microliters of supernatants were diluted 3-fold with distilled water and subjected to high performance liquid chromatography on an Agilent 1100 HPLC system using a reversed phase C-18 column (Nucleodur Sphinx RP, 250 × 4.6 mm, 5 µm, Macherey-Nagel, Düren, Germany) with a water (A)/acetonitrile (B) gradient (1 min: 1.5% B; 5 min: 1.5–5% B; 2 min: 5–7% B; 10 min: 7–21% B; 5 min, 21–29% B; 0.1 min: 29–100% B; 0.9 min: 100% B; 4 min: 1.5% B; flow rate: 1.0 ml min−1). Detection was performed with a photodiode array detector and peaks were integrated at 229 nm. Desulfoglucosinolates were identified based on ultraviolet absorption spectra, retention time, and mass spectra from liquid chromatography-mass spectrometry (LC-MS) analysis conducted with a Bruker Esquire 6000 IonTrap mass spectrometer.
Esquire 6000 iontrap mass spectrometer
Manufactured by Bruker
The Esquire 6000 IonTrap mass spectrometer is a laboratory instrument designed for the analysis of chemical compounds. It utilizes ion trap technology to capture, isolate, and detect ions based on their mass-to-charge ratio. The core function of the Esquire 6000 is to provide accurate mass measurements and facilitate the identification of unknown substances.
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Lab products found in correlation
2 protocols using esquire 6000 iontrap mass spectrometer
1
Quantification of Desulfoglucosinolates via HPLC
2
Purification and Characterization of Organic Compounds
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Unless otherwise noted, materials were obtained from co mme rcial suppliers and used without further purification. All reactions requiring anhydrous conditions were conducted in oven-dried glassware under a dry nitrogen atmosphere. The solvents were re moved under reduced pressure with a rotary evaporator. Residues were purified by chromatography on a silica gel column (230 -400 mesh) by using mixtures of hexane/ethyl acetate as the eluent. 1 H and 13 C NMR spectra were measured on a 400 or a 300 MHz NM R spectrometer . 1 H and 13 C che mical shifts (δ) are re ferenced to internal solvent resonances and reported relative to SiMe4. The che mical shifts were assigned on the basis of 2D COSY, NOESY, DOSY, 1 H-13 C HSQC , 1 H-13 C HMB C and 1 H-31 P HMBC experi ments per for med under routine conditions. Electrospray mass spectrometry analyses were recorded on an Esquire 6000 ion trap mass spectrometer (Bruker) equipped with an electrospray ion source, operated in positive ESI(+) ion mode. IR spectra were recorded with an FT-IR using a single reflection ATR system as a sampling accessory.
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