The NMR spectra were recorded with a Bruker Avance 500 MHz spectrometer. The high resolution mass spectra were recorded with a Bruker Daltonics microTOF-Q instrument. The LC-MS chromatograms were recorded with a Agilent Technologies 1260 Infinity Binary LC and Agilent 6100 Series Quadrupole LC/MS Systems with Phenomemex 150 × 4.6 SynergiTM 4 μm Fusion-RP 80 Å analytical column (flow rate 0.5 ml/min and wavelength 260 nm, 0.1% formic acid in H2O and MeCN as eluents).
1260 infinity binary lc
Manufactured by Agilent Technologies
Sourced in United States
The 1260 Infinity Binary LC is a high-performance liquid chromatography (HPLC) system designed for analytical applications. It features a binary pump that can deliver a precise and stable solvent flow, allowing for reliable and reproducible separation of analytes.
Automatically generated - may contain errors
Lab products found in correlation
Superdex 75 prep grade gel, by GE Healthcare (2 mentions)
6230 accurate mass tofms, by Agilent Technologies (2 mentions)
Microtof q instrument, by Bruker (1 mentions)
Avance 500 mhz spectrometer, by Bruker (1 mentions)
Nucleoside digestion mix, by New England Biolabs (1 mentions)
Q exactive hybrid quadrupole orbitrap mass spectrometer, by Thermo Fisher Scientific (1 mentions)
Zorbax hilic plus column, by Agilent Technologies (1 mentions)
6420 triple quadrupole lc ms, by Agilent Technologies (1 mentions)
Speedvac vacuum concentrator kit, by Thermo Fisher Scientific (1 mentions)
Beh c18 column, by Waters Corporation (1 mentions)
8 protocols using 1260 infinity binary lc
1
NMR, HRMS, and LC-MS Analysis
2
Quantitative Analysis of Bacterial Genomic Nucleosides
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Nucleoside Digestion Mix (New England Biolabs, Ipswich, MA, cat#M0649S) was used to digest YerA41 genomic material into nucleosides. Briefly, 50 μl reactions were prepared with 2 μl of the enzyme mix, 5 μl of the 10× Nucleoside Digestion Mix buffer and 43 μl of YerA41 genomic material. Samples were incubated overnight at 37°C and stored at 4°C. The LC-MS chromatograms of YerA41 genomic samples digested with the Nucleoside Digestion Mix were recorded with a Agilent Technologies 1260 Infinity Binary LC and Agilent 6100 Series Quadrupole LC/MS Systems with Phenomemex 150 × 4.6 SynergiTM 4 μm Fusion-RP 80 Å analytical column at the University of Turku, Finland. The nucleoside samples were eluted for 2.5 min with 1% MeCN and 99% formic acid (0.1%) buffer in mQ water, and then increasing MeCN volume linearly to 10% at 25 min (25 (link)). Tandem mass spectroscopy (LC–MS/MS) was recorded with Thermo Scientific Q Exactive Hybrid Quadrupole-Orbitrap Mass Spectrometer using HPLC conditions mentioned above.
3
Mechanistic probe acylation of SalB-PCP
4
HPLC Analysis of Phytochemical Profiles
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Phytochemical profiles of both red and white NAE and NEE were screened by high-performance liquid chromatography (HPLC)-diode array (Agilent 1260 Infinity Binary LC, Santa Clara, CA, USA) [46 (link)]. The HPLC condition comprised of Purospher® Star PR-18 endcapped column (150 × 4.60, 5 µm). The mobile phase consists of 92% A (0.1% formic acid in water) and 8% B (acetonitrile), maintained for 10 min. The B was increased to 14% in 24 min, 23% in 35 min and 24% in 60 min. Then, 10 µL of the sample was injected, and the spectra were determined at 250 nm and 330 nm [47 ]. The spectra between 200 and 400 nm were collected. The identification of the chromatographic peak was achieved by comparing the retention times and spectral characteristics of the eluted peaks with the standards.
5
Betaine and Proline Quantification by LC-MS/MS
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Betaine and proline in intra/extracellular metabolites was analyzed by LC–MS/MS under multiple reaction monitoring (MRM) methods. Ten microliters of each prepared sample was analyzed by an Agilent 6420 Triple Quadrupole LC/MS (CA, USA) coupled to an Agilent 1260 Infinity Binary LC (CA, USA). Separations were performed using an Agilent Zorbax HILIC Plus column (4.6 × 100 mm, 3.5 μm) and a flow rate of 500 μL min−1. Eluent A was water containing 0.1% v/v formic acid, and B was 100% acetonitrile. The gradient applied was as follows: 0–3 min, 80% eluent B; 3–10 min, linear decrease to 20% eluent B; 10–13 min, 20% eluent B; 13–14 min, linear increase to 80% eluent B; 14–23 min, 80% eluent B. The flow rate was at 0.5 mL min−1. The capillary temperature was 300 °C, and an electrospray ionization spray voltage was used at 4 kV.
6
Offline Phosphopeptide Fractionation and Vacuum Concentration
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The TMT-labeled phosphopeptides were loaded onto a 4.6 mm inner diameter 150 mm outer diameter BEH C18 column (Waters Corp) for offline fractionation into 16 fractions. The peptides were separated using a high pH reversed-phase gradient (10 mM ammonium formate Mobile Phase A, 10 mM ammonium formate in 80% MeOH Mobile Phase B) on an Agilent 1260 Infinity Binary LC equipped with an automatic fraction collector. The effective separation gradient started at 20% Mobile Phase B and increased to 60% Mobile Phase B in 6 min, flowing at a rate of 800 μL/min. The fractions were then concatenated into 8 fractions, combining fractions 1 and 9, 2 and 10, 3 and 11, 4 and 12, 5 and 13, 6 and 14, 7 and 15, and 8 and 16. The combined fractions were transferred to 2 mL Starstedt micro tubes for rapid evaporation under vacuum with the SpeedVac Vacuum Concentrator Kit (Thermo).
7
Mechanistic probe acylation of SalB-PCP
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SalB-PCP was acylated with the panthetheine-activated mechanistic probe 21 using the same methods described earlier. This holo-acyl-SalB-PCP was purified away from residual CoA proteins and small molecules using a Superdex 75 prep grade gel filtration column (16 cm x 60 cm, GE Healthcare) preequilibrated with buffer (50 mM Tris, 150 mM NaCl, 5% glycerol, pH 8.0). Purified 22 or diffusible substrates (21–25 ) were incubated with SalC or a SalC variant (20 μM) for 3 h, and then subjected to LC-HRMS analysis. All intact protein LC-HRMS was performed by the UC San Diego Molecular Mass Spectrometry Facility on an Agilent 1260 Infinity Binary LC coupled with a 6230 Accurate-Mass TOFMS.
8
Phytochemical Profiling of M. oleifera Leaf Tea
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M. oleifera leaf tea was screened for phytochemical profiles with a high-performance liquid chromatography (HPLC)-diode array (Agilent 1260 Infinity Binary LC, Santa Clara, CA, USA) [53 (link)]. The HPLC condition was comprised of Purospher® Star PR-18 endcapped column (150 × 4.60, 5 µm). The mobile phase consisted of 92% A (0.1% formic acid in water) and 8% B (acetonitrile), maintained for 10 min. Then B was increased to 14% in 24 min, to 23% in 35 min, to 24% in 60 min and the sample injection volume was 10 µL. The spectra were determined at 250 nm and 330 nm. The spectra between 200 to 400 nm were collected and identified. The chromatographic peak was achieved by comparing the retention times and spectral characteristics of the eluted peaks with the standards.
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