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6530bq tof mass spectrometer

Manufactured by Agilent Technologies

The 6530BQ-TOF mass spectrometer is a high-performance quadrupole time-of-flight (Q-TOF) mass spectrometer designed for accurate mass measurements and high-resolution analysis. It features a quadrupole mass analyzer and a time-of-flight mass analyzer to provide precise mass determination and detailed structural information about analytes.

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3 protocols using 6530bq tof mass spectrometer

1

Synthesis and Characterization of Pyridyl Ligands

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All reactions were carried out under
an oxygen-free, N2 atmosphere using standard Schlenk line
techniques. The starting materials were purchased from Alfa-Aesar
and Sigma-Aldrich Chemicals. Mn2(CO)10, trimethylamine-N-oxide, isonicotinoyl chloride hydrochloride, 1,2-ethanediol,
1,2-dihydroxybenzene, diethylene glycol, 1,2-ethylene diamine, 1,4-phenylenediamine, p-benzoquinone, n-butylamine, and phenethylamine
were used as received. The aminoquinone ligands (bbbq and bpbq) and
ditopic pyridyl ligands (etdp, pcadgd, pdi, bpce, and pdia) were synthesized
as reported in the literature.17 (link) Dichloromethane,
ethanol, methanol, tetrahydrofuran, and other solvents were dried
using standard methods and freshly distilled prior to use.18 IR spectra were recorded on a Nicolet iS10 Fourier
transform infrared spectrometer. Electronic absorption spectra were
obtained on a Shimadzu UV-2450 spectrophotometer. Emission spectra
were recorded on a Fluoromax-4 spectrofluorometer. Solvents used for
UV–vis and emission titration experiments were of spectral
grade. 1H NMR spectra were recorded on a Bruker Avance
400 MHz NMR spectrometer with tetramethylsilane as the internal reference.
Elemental analyses were performed using a Thermo Scientific Flash
2000 CHNS analyzer. ESI-mass spectra were taken on an Agilent 6530B
Q-TOF mass spectrometer.
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2

Quantitative Analysis of RNA Samples

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RNA samples in TE buffer (3 uM) were analyzed by mass spectrometry (Agilent 1290 Infinity II liquid chromatography system (LC) coupled with Agilent 6530B Q-TOF mass spectrometer (MS)) in a negative ion polarity mode. LC is performed with gradient elution (buffer A: 50 mM HFIP; 15 mM Hexylamine 2% MeOH; buffer B: MeOH, 0.75 mL/min, 2–95% B in 1.05 min) on an Acquity UPLC BEH C18 VanGuard Pre-column (1.7 um, 2.1 × 5 mm). Electrospray ionization performed with a dual ESI source (gas temp 325 °C, drying gas 12 L/min, nebulizer 40 psi, Vcap 4 kV, fragmentor 250, skimmer 65). Data acquired in 100–3200 m/z range and deconvoluted in 4000–40000 m/z range.
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3

Lipid Extraction and LC-MS Lipidomics

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Serum aliquots were extracted as previously described (28 (link)). The top layer of the 2-phase solution was used for lipidomic analyses. Extracts were analyzed on an Agilent 1290 Infinity II LC System (Santa Clara, CA) with a 100 mm long, 2.1 mm id, and 1.7 μm particles Waters Acquity UPLC CSH C18 column protected by a short guard column. Mass spectrometry was performed using an Agilent 6530 QTOF mass spectrometer with resolution R=10,000 for positively charged lipids and an Agilent 6530b QTOF mass spectrometer with resolution R=20,000 for negatively charged lipids. Twenty-four internal standards, optimized for lipidomics, were added to samples for retention time alignment markers, quality control purposes, and quantification corrections.
Raw data were processed using MS-DIAL (version 2.8) (29 (link)). Quantification of metabolites are reported as peak heights. All samples were analyzed in one batch, and data quality and instrument performance were constantly monitored using quality controls, which were comprised of pooled plasma samples (BioIVT) and injected every 10 samples.
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