All reagents were obtained from commercial sources and used without further purification. TCO-PEG3-Amine, TCO-PEG4-NHS, Methyltetrazine-PEG4-NHS, and tetrazine-fluorophores (AF488, Cy3, AF594) were purchased from Click Chemistry Tools, LLC. Flash column chromatography was performed using Sorbtech purity flash cartridges or Biotage SNAP Bio C18 columns for reversed phase chromatography. NMR spectra were recorded on a Bruker Avance UltraShield 400 MHz spectrometer. Chemical shifts are reported in parts per million (δ) and referenced to the residual solvent. Reactions were monitored via liquid chromatography-mass spectrometry (LC-MS) on a Waters instrument equipped with a Waters 2424 ELS Detector, Waters 2998 UV-Vis Diode array Detector, and a Waters 3100 Mass Detector. UV-Vis analysis of antibodies was performed on a NanoDrop 1000 spectrophotometer.
Waters 2424 els detector
Manufactured by Waters Corporation
Sourced in United Kingdom
The Waters 2424 ELS Detector is a laboratory instrument designed to detect and analyze various compounds in liquid chromatography (LC) and other analytical applications. The core function of the 2424 ELS Detector is to provide sensitive and selective detection of analytes that do not possess a chromophore, allowing for their quantification and identification.
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Lab products found in correlation
Waters 2998 uv vis diode array detector, by Waters Corporation (4 mentions)
3100 mass detector, by Waters Corporation (3 mentions)
Xbridge beh c18 column, by Waters Corporation (2 mentions)
Avance ultrashield 400 mhz spectrometer, by Bruker (1 mentions)
Tco peg4 nhs, by Vector Laboratories (1 mentions)
2795 separations module, by Waters Corporation (1 mentions)
2996 photodiode array detector, by Waters Corporation (1 mentions)
Waters 3100 mass detector, by Waters Corporation (1 mentions)
Xterra ms c18 column, by Waters Corporation (1 mentions)
Ascend 400 mhz spectrometer, by Bruker (1 mentions)
2475 multi wavelength fluorescence detector, by Waters Corporation (1 mentions)
Microflex maldi tof ms, by Bruker (1 mentions)
Dual fl spectrophotometer, by Horiba (1 mentions)
6 protocols using waters 2424 els detector
1
Synthesis and Characterization of Clickable Reagents
2
Quantification of Acylglycerols by HPLC-ELSD
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The FFA and monoacylglycerol (MAG) to DAG ratio as well as the 1,3-DAG to 1,2-DAG ratio were analyzed according to the AOCS Official Method Cd 11-96 using an HPLC system (Alliance model Waters 22.695 Separation Modules, Wilmslow, UK) equipped with an ELSD detector (Alliance model Waters 2424 ELS Detector, Wilmslow, UK).
The test sample was diluted in a mixture of n-hexane and isopropanol (ratio 9:1). The diluted sample was filtered through a PTFE membrane filter and injected into a normal phase silica column (150 mm × 4.6 mm, LiChrospher ® 60 Si (10 µm), Hibar, Merck, Selangor, Malaysia). The injection volume was 20 µL. Eluent A (n-hexane) and eluent B (n-hexane:ethyl acetate:isopropanol at an 8:1:1 ratio) were used to establish a gradient program. The gradient started with 98% of eluent A and 2% of eluent B and reached 65% eluent A and 35% of eluent B at 8 min; then, 2% of eluent A and 98% of eluent B at 8.5 min, held for 15 min; and lastly, 98% of eluent A and 2% of eluent B at 15.1 min, held until 19 min. Total flow rate of the eluent was 2 mL/min. The column oven temperature was 40 °C, while the drift tube and temperature of the detector was 60 and 36 °C, respectively. Nitrogen gas flow was constant at 40 psi.
The test sample was diluted in a mixture of n-hexane and isopropanol (ratio 9:1). The diluted sample was filtered through a PTFE membrane filter and injected into a normal phase silica column (150 mm × 4.6 mm, LiChrospher ® 60 Si (10 µm), Hibar, Merck, Selangor, Malaysia). The injection volume was 20 µL. Eluent A (n-hexane) and eluent B (n-hexane:ethyl acetate:isopropanol at an 8:1:1 ratio) were used to establish a gradient program. The gradient started with 98% of eluent A and 2% of eluent B and reached 65% eluent A and 35% of eluent B at 8 min; then, 2% of eluent A and 98% of eluent B at 8.5 min, held for 15 min; and lastly, 98% of eluent A and 2% of eluent B at 15.1 min, held until 19 min. Total flow rate of the eluent was 2 mL/min. The column oven temperature was 40 °C, while the drift tube and temperature of the detector was 60 and 36 °C, respectively. Nitrogen gas flow was constant at 40 psi.
3
Synthesis and Characterization of ISRIB Analogs
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Commercially available reagents and solvents were used as received. Compounds ISRIB-A1 and ISRIB-A2 were prepared as previously reported (Sidrauski et al., 2013b). Compound ISRIB-A7 was available commercially from Specs (The Netherlands). 1H NMR spectra were recorded on a Varian INOVA-400 400 MHz spectrometer and a Bruker Avance 300 300 MHz spectrometer. Chemical shifts are reported in δ units (ppm) relative to residual solvent peak. Coupling constants (J) are reported in hertz (Hz). LC-MS analyses were carried out using Waters 2795 separations module equipped with Waters 2996 photodiode array detector, Waters 2424 ELS detector, Waters micromass ZQ single quadropole mass detector, and an XBridge C18 column (5 µm, 4.6 × 50 mm). Microwave reactions were carried out in a CEM Discover microwave reactor.
4
NMR Spectroscopy and HPLC-MS Analysis
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NMR spectra were recorded on a Bruker Avance UltraShield 400 MHz spectrometer. 1H NMR chemical shifts were reported in ppm relative to SiMe4 (δ = 0) and were referenced internally concerning residual protons (δ = 4.79 for D2O). Peak assignments, calculated chemical shifts, and peak integrals were based on reference solvent peaks. 2D Rotating Frame Overhauser Enhancement Spectroscopy (2D‐ROESY) experiments were performed to assess the interactions between dipolarly coupled hydrogens, and integrals were normalized to the reference hydrogen (H1) of the sbCD. All experiments were performed in D2O (0.5 mL) at a fixed sCD concentration (26 mM) with 10% (CD3)2SO. High‐performance liquid chromatography‐mass spectrometry analysis (HPLCMS) was performed on a Waters instrument equipped with a Waters 2424 ELS Detector, Waters 2998 UV–Vis Diode array Detector, and a Waters 3100 Mass Detector. Separations employed an HPLC‐grade water/acetonitrile (0.1% formic acid) solvent gradient with XTerra MS C18 Column, 125 Å, 5 µm, 4.6 × 50 mm column; Waters XBridge BEH C18 Column, 130 Å, 3.5 µm, 4.6 × 50 mm.
5
NMR and HPLC-MS Characterization of Cyclodextrin Complexes
6
Comprehensive Analytical Characterization
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