The NanoLC-MS/MS was performed with a nanoflow UPLC system (Ultimate 3000, RSLCnano system, Dionex, Amsterdam, Netherland) coupled with coupled with a captive spray ion source and a Q-TOF mass spectrometer (maXis impact II, Bruker). The samples were injected into a tunnel frit trap column (C18, 5 μC, 100 Å, packed length of 2 cm, 375 μ od × 180 μ × id) [2 (link)] with a flow rate of 10 μl/min and then separated by an analytical column (Acclaim PepMap C18, 2 μm, 100 Å, 75 μm × 250 mm, Thermo Scientific, USA) with a flow rate of 300 nl/min. A gradient elution of 5% ACN (0.1% FA) to 35% ACN (0.1%) within 80 min was used for peptide separation. Eight precursors of charge +2, +3 and +4 from each TOF-MS scan (400–2000m/z) were dynamically selected and isolated for MS/MS fragment ion scan (50–2000m/z). Selected precursors were then actively excluded for 20 s. MS and MS/MS accumulation were set at 1 and 2 Hz.
Maxis impact 2
Manufactured by Bruker
Sourced in Germany
The Maxis Impact II is a high-performance mass spectrometer designed for accurate and sensitive analysis of a wide range of samples. It features a robust and reliable design that enables efficient data acquisition and processing. The Maxis Impact II is a versatile instrument that can be used for various applications in analytical chemistry and life sciences.
Automatically generated - may contain errors
Lab products found in correlation
Acclaim pepmap c18, by Thermo Fisher Scientific (2 mentions)
Ultimate 3000 rslcnano system, by Thermo Fisher Scientific (1 mentions)
Ods am, by YMC (1 mentions)
Lc 20, by Shimadzu (1 mentions)
Chirascan plus cd spectrometer, by Applied Photophysics (1 mentions)
Hydro rp, by Phenomenex (1 mentions)
Si gel60 rp 18 f254s, by Merck Group (1 mentions)
Uv 1601pc spectrometer, by Shimadzu (1 mentions)
Avance 3 700, by Bruker (1 mentions)
Agilent 1100, by Agilent Technologies (1 mentions)
Avance 300, by Bruker (1 mentions)
Avance 500, by Bruker (1 mentions)
Equinox 55 fourier transform, by Bruker (1 mentions)
Avance drx 500, by Bruker (1 mentions)
Chirascan plus quick start cd spectrometer, by Applied Photophysics (1 mentions)
Acclaim pepmap rslc column, by Thermo Fisher Scientific (1 mentions)
Tasq 1, by Bruker (1 mentions)
5 protocols using maxis impact 2
1
NanoLC-MS/MS Proteomic Analysis
2
Spectroscopic Characterization of Organic Compounds
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Optical rotations were measured on a Perkin-Elmer 343 polarimeter in MeOH. UV spectra were recorded on a Shimadzu UV-1601PC spectrometer in MeOH. ECD spectra were measured on a Chirascan-Plus CD Spectrometer in MeOH. 1H and 13C NMR spectra were recorded in aceton-d6 on a Bruker Avance-300, Avance-500 and Avance III-700 spectrometers (Bruker BioSpin GmbH) operating at 300 and 75, 500 and 125 MHz and 700 and 176 MHz, respectively. HRESIMS spectra were obtained on a Bruker maXis Impact II mass spectrometer (Bruker Daltonics GmbH). Low-pressure liquid column chromatography was performed using C18-SiO2 Gel ODS-A (12 nm, S—75 um, YMC Co., Ishikawa, Japan) and a Buchi B-688 Chromatography Pump on a Buchi glass column using Si gel KSK (50/100 μm, Imid Ltd., Russia). Plates precoated with Si gel (5–17 μm, 4.5 × 6.0 cm, Imid Ltd., Russia) and Si gel 60 RP-18 F254S (20 × 20 cm, Merck KGaA, Germany) were used for thin-layer chromatography. Preparative HPLC was carried out on a Shimadzu LC-20 (Shimadzu, Kyoto, Japan) and Agilent 1100 (Agilent Technologies, Santa Clara, CA, USA) chromatographs using a Shimadzu RID-20A and Agilent 1100 refractometers and YMC ODS-AM (YMC Co, 5 μm, 250 × 10 mm), Ultrasphere Si (5 μm, 250 × 4.6 mm), and Hydro-RP (Phenomenex, 4 μm, 250 × 10 mm) columns.
3
Spectroscopic Analysis of Organic Compounds
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Infrared (IR) spectra were obtained on an Equinox 55 Fourier-transform (FT) IR spectrophotometer (Bruker, Rheinstetten, Germany). The CD spectra were obtained on a Chirascan-plus Quick Start CD Spectrometer (Applied Photophysics Limited, Leatherhead, UK) (acetonitrile, 20 °C). The 1H-, 13C- and two-dimensional (2D) NMR spectra were recorded in DMSO-d6 using an NMR Bruker AVANCE DRX-500 instrument (Bruker, Karlsruhe, Germany). The chemical shift values (δ) and the coupling constants (J) are given in parts per million and Hz, respectively. HMBC spectra were optimized for 5 Hz coupling. HRESIMS spectra were obtained on a Bruker Maxis Impact II instrument (Bruker Daltonics, Bremen, Germany).
4
Protein Purification and Identification
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The enzymatic extract was concentrated on a PierceTM Protein Concentrator (10K MWCO: 10 kDa molecular weight cutoff) by 50-fold. Electrophoretic separation of proteins was performed by SDS-PAGE in a 12% polyacrylamide gel. The proteins were extracted from the gel samples and trypsinized. Peptides were separated on an Acclaim Pep-Map RSLC column (Thermo Scientific, Waltham, MA, USA) with an acetonitrile gradient. An unmodified CaptiveSpray ion source was employed to interface the HPLC system with a Maxis Impact II instrument (Bruker). The mass range of the MS scan was set to m/z 150–2200 in positive ion polarity mode. The Mascot software was used to perform searches against the SwissProt 2016_08 database (552,884 sequences; 197,760,918 residues).
5
Quantitative Pesticide Screening Using UHPLC-QTOF-MS
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Chromatographic separation was achieved with a Dionex Ultimate 3000 UHPLC system containing a Dionex Acclaim RSLC 120 C 18 analytical column (2.1 i.d. × 100 mm length, 2.2 μm particle size), (Thermo Fisher Scientific, Bremen, Germany) and a VanGuard, Acquity UPLC BEH C18 guard column (1.7 μm particle size), (Waters, Dublin, Ireland). Mass spectrometry was undertaken with a Bruker Maxis Impact II electrospray high resolution time-of-flight tandem mass spectrometer (Q-TOF-MS) (Bruker Daltonics, Bremen, Germany), with the following operating parameters: capillary voltage, 2500 V; end plate offset, 500 V; nebulizer pressure, 2 bar (N2); drying gas, 8 L min -1 (N2); and drying temperature, 200 °C. Data acquisition used Bruker HyStar acquisition software (rev. 3.2) and data interpretation (analyte identification and quantitation) used Bruker Target Analysis for Screening and Quantitation (TASQ ® ) 1.4 software. Software and hardware used for chromatographic separation, mass spectrometry, data acquisition and data interpretation where interfaced and used to screen against Bruker's PesticideScreener ™ (2.1) database. Statistical interpretation of identified analytes was undertaken after transferring outputs contained in TASQ ® into Mass Profiler Professional (MPP) software (B.14.9.1) (Agilent, Santa Clara, USA).
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