Microtof q 3 spectrometer

Manufactured by Bruker

Sourced in United States

The MicrOTOF-Q III is a high-performance quadrupole time-of-flight mass spectrometer (QTOF-MS) designed for analytical applications. It combines the high resolution and mass accuracy of a time-of-flight (TOF) analyzer with the versatility of a quadrupole mass filter. The instrument is capable of performing accurate mass measurements and is suitable for a wide range of analytical techniques, including liquid chromatography-mass spectrometry (LC-MS) and direct infusion mass spectrometry.

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7 protocols using microtof q 3 spectrometer

Analytical Characterization of Compounds

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The NMR spectra were recorded using Bruker BioSpin AV-300 and AV-600 spectrometers. The ESI-MS spectra were recorded using a Bruker Daltonics micrOTOF Q-III spectrometer (Bruker Daltonics, Billerica, MA, USA). The HPLC and GPC measurements were conducted using a system consisting of a JASCO PU-2089 pump and a JASCO CO-2065 column oven (JASCO Corporation, Tokyo, Japan). A JASCO UV-2075 ultraviolet detector and a JASCO RI-2031 refractive index detector were used for the HPLC and GPC analyses, respectively. A 5C18-MS-II column (ɸ4.6 × 250 mm, Nacalai Tesque, INC.) was used for the HPLC analysis. 5, 4, 3, and 10 % MeCN-containing water were used as the eluents at a flow rate of 1.0 mL/min at 30 °C to analyze the enzymatic reaction with 1a – d , respectively. A Shodex OHpak SB-804 HQ column (ɸ8.0 × 300 mm, Showa Denko K.K., Tokyo, Japan) was used for the GPC analysis of 3a – c using a phosphate buffer (20 mM, pH 7.0) as the eluent at a flow rate of 0.5 mL/min at 30 °C. Pullulan samples were used as standards. A Shodex KD-804 column (ɸ8.0 × 300 mm, Showa Denko K.K.) was used for the GPC analysis of 3d using N , N -dimethylformamide (DMF) containing 10 mM lithium bromide as the eluent at a flow rate of 0.5 mL/min at 50 °C. Poly(methylmethacrylate) samples were used as standards. The fluorescence intensity was recorded using a JASCO FP-6500 fluorometer for the lectin binding tests.

Tanaka T., Matsuura A., Aso Y, & Ohara H. (2020). Aqueous One-pot Synthesis of Glycopolymers by Glycosidase-catalyzed Glycomonomer Synthesis Using 4,6-Dimetoxy Triazinyl Glycoside Followed by Radical Polymerization. Journal of Applied Glycoscience, 67(4), 119-127.

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NMR and Mass Spectrometry of Alkaloids

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¹H vand ¹³C NMR spectra were performed in hexadeuterodimethylsulphoxide (DMSOd6) on a Brucker DRX 500 spectrometer at Centro de Química, IVIC. Mass spectra were measured in a Bruker Micro TOF-QIII spectrometer set to ESI mode using MeOH as the solvent at Centro de Biología Estructural, IVIC. NMR spectra of norpurpureine and purpureine (Additional file 1) and MS spectra of norpurpureine and purpureine (Additional file 2), are available.

Sánchez G., Estrada O., Acha G., Cardozo A., Peña F., Ruiz M.C., Michelangeli F, & Alvarado-Castillo C. (2018). The norpurpureine alkaloid from Annona purpurea inhibits human platelet activation in vitro. Cellular & Molecular Biology Letters, 23, 15.

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Insulin Aggregation Characterization

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Human recombinant insulin (Sigma-Aldrich, Poznan, Poland), essentially fatty acid free human serum albumin (HSA) (Sigma-Aldrich), 4-(1-pyrenyl)butyric acid (Merck, Warsaw, Poland), Human recombinant insulin (Sigma-Aldrich), Congo Red, Thioflavin T (Sigma-Aldrich, Poznan, Poland), and all necessary amino acid derivatives (Merck) were used as received. NaH₂PO₄∙2H₂O, Na₂HPO₄∙12H₂O, TX-100, EtOH, MeOH, and NaCl (Sigma-Aldrich) were used for the buffer preparation. Water was purified with the use of a Millipore Milli-Q Plus system.
Analytical HPLC: UltiMate 3000 UHPLC System Thermo Scientific™; column parameters: Kinetex 2.6 u C18 100A, 100 × 4.6 mm, 20 °C; diode array UV/Vis detector (DAD); eluent ACN/H₂O; gradient 0–2 min 3/97, 2–31 min 95/5, 31–32 min 0/100, 32–33 min 0/100, 33–35 min 3/97, 35–37.5 min 3/97.
Preparative HPLC: CombiFlash, EZPrep, Teledyne ISCO, Supelco Discovery BIO Wide Pore C18 column (25 cm × 21.2 mm, 10 mm; Sigma Aldrich); flow rate, 5 mL/min; detection wavelengths, 220 and 254 nm); gradient ratio A (0.1% TFA in ACN)/ B (0.1% TFA in H₂O) 0:100 to 18:82 in 30 min, followed by an isocratic run for 5 min.
ESI/MS: micrOTOF-Q III spectrometer Bruker Daltonics equipped with electrospray source (ESI) and time of flight detector (TOF).

Wasko J., Wolszczak M., Kaminski Z.J., Steblecka M, & Kolesinska B. (2020). Human Serum Albumin Binds Native Insulin and Aggregable Insulin Fragments and Inhibits Their Aggregation. Biomolecules, 10(10), 1366.

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Characterization of Polymer Samples

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NMR spectra were recorded using a Bruker BioSpin AV-300 spectrometer. ESI mass spectra were recorded using a Bruker Daltonics microTOF Q-III spectrometer. GPC measurements were conducted using a system consisting of a JASCO PU-2089 pump, a CO-2065 column oven, an RI-2031 refractive index detector, and a Shodex OHpak SB-804 HQ (8.0 × 300 mm) column. 20 mM phosphate buffer (pH 7.0) was used as the eluent at a flow rate of 0.5 mL/min at 30 ˚C. Pullulan samples were used as standards.

Tanaka T., Zhou Y., Tamoto C., Kurebayashi Y., Takahashi T, & Suzuki T. (2017). An α2,3-Linked Sialylglycopolymer as a Multivalent Glycoligand Against Avian and Human Influenza Viruses. Journal of Applied Glycoscience, 64(2), 43-48.

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Synthesis of Organic Compounds

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Chemical reagents used were commercially available (Tedia, Applichem, Chem-Impex International, Sigma Aldrich, Oakwood Products, Lancaster Avocado, Alfa-Aesar, Fisher). All reactions were conducted with magnetic stirring under an argon atmosphere in oven-dried flasks. Reactions were monitored until deemed complete by TLC using silica-gel-coated glass plates (Merck Kiselgel 60 F254). Plates were visualized under UV light (254nm). Plates were dyed with 10% phosphomolybdic acid (PMA) in ethanol. ¹H, and ¹³C NMR spectra were recorded at 500 (¹H), and 125MHz (¹³C) on an Agilent Inova 500 spectrometer; and at 400 (¹H), 100MHz (¹³C) on Eclipse 400MHz spectrometer (JEOL, Peabody, MA, USA). Chemical shifts (δ) are reported in parts per million (ppm) using the residual solvent peak and coupling constants (J) are given in Hz. Proton multiplicity is reported as singlet (s), doublet (d), triplet (t), quartet (quart.), quintet (quint.), septet (sept), multiplet (m), and broad (br). Infrared spectrophotometry was carried out on a Platinum ATR Alpha instrument (Brucker, Billerica, MA, USA). The molar masses were determined with a micrOTOF-QIII spectrometer Bruker Daltonics, Billerica, MA, USA), with electrospray ionization (ESI) and positive ion detection mode. The detailed synthetic procedures and spectral characterization are described below.

Lakey-Beitia J., González Y., Doens D., Stephens D.E., Santamaría R., Murillo E., Gutiérrez M., Fernández P.L., Rao K.S., Larionov O.V, & Durant-Archibold A.A. (2017). Assessment of Novel Curcumin Derivatives as Potent Inhibitors of Inflammation and Amyloid-β Aggregation in Alzheimer's Disease. Journal of Alzheimer's disease : JAD, 60(Suppl 1), S59-S68.

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Analytical Techniques for Chemical Characterization

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Nuclear magnetic resonance (NMR) analyses for ¹H and ¹³C NMR spectra were measured on either a JEOL AL300 (300 MHz) or an AL400 (400 MHz) instrument. High-resolution mass spectroscopy (HRMS) was carried out on a Bruker MicrOTOF-QIII spectrometer by electrospray ionization time-of-flight (ESI-TOF). Matrix-assisted laser desorption ionization (MALDI-TOF) mass spectrometry analysis was obtained on a Bruker autoflex spectrometer using 2,5-dihydroxybenzoic acid as a matrix. Reversed-phase high-performance liquid chromatography (HPLC) was performed using a Shimadzu system (Kyoto, Japan), consisting of two LC-20AP pumps and a SPD-20AV photodiode array detector. The used column was a preparative 20 × 250–mm Cosmosil 5C₁₈-AR-300 from Waters Corporation (MA, USA). Ultrapure water used for all experiments described in this paper was obtained from the Milli-Q Advantage A10 Water Purification System sold by Merck Millipore (Burlington, USA).

Vong K., Tahara T., Urano S., Nasibullin I., Tsubokura K., Nakao Y., Kurbangalieva A., Onoe H., Watanabe Y, & Tanaka K. (2021). Disrupting tumor onset and growth via selective cell tagging (SeCT) therapy. Science Advances, 7(17), eabg4038.

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Solvent Purification and Moisture-Sensitive Reactions

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Solvents were distilled prior to use. Anhydrous DMF, THF, and DCM were obtained from a solvent purification system, or purchased from Merck-Sigma Aldrich (Taufkirchen, Germany). Moisture-sensitive reactions were performed under a nitrogen atmosphere. Reactions were monitored by analytical thin-layer chromatography, using precoated silica gel plates with an ultraviolet (254 nm) indicator (ALUGRAM^® Xtra SIL G/UV₂₅₄ from Macherey-Nagel, Düren, Germany). Flash column chromatography was performed with silica gel (Macherey-Nagel, Silica 60, particle size 0.040–0.063 mm). NMR spectra were measured at 295 K on a Bruker AVANCE 400 FT-NMR spectrometer. Deuterated solvents were used. The chemical shifts are reported in parts per million (ppm, δ-scale), referring to the residual solvent peak (¹H: CDCl₃: δ = 7.26 ppm; ¹³C: CDCl₃: δ = 77.1 ppm). Analysis followed first order, and data are reported in the following order: chemical shift (in ppm), multiplicity (s = singlet, d = doublet, dd = doublet of doublets, t = triplet, tt = triplet of triplets, dt = doublet of triplets, q = quartet, m = multiplet, br m = broad multiplet), coupling constant (s) (in Hz), and integration. Mass spectra were recorded with a Bruker Daltonics (Billerica, MA, USA) micrOTOF-Q III spectrometer by electrospray ionization (ESI).

Geiger N., Kersting L., Schlegel J., Stelz L., Fähr S., Diesendorf V., Roll V., Sostmann M., König E.M., Reinhard S., Brenner D., Schneider-Schaulies S., Sauer M., Seibel J, & Bodem J. (2022). The Acid Ceramidase Is a SARS-CoV-2 Host Factor. Cells, 11(16), 2532.

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