Uplc ms system

Manufactured by Waters Corporation

The UPLC-MS system is a high-performance liquid chromatography and mass spectrometry instrument developed by Waters Corporation. It is designed to provide rapid and efficient separation and analysis of complex samples. The UPLC-MS system combines the ultra-high-pressure liquid chromatography (UPLC) technology with a mass spectrometer, enabling sensitive and selective detection of analytes in a variety of applications.

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Lab products found in correlation

Acquity pda detector, by Waters Corporation (1 mentions) Jumpstart taq dna polymerase, by Merck Group (1 mentions) Doxorubicin, by Selleck Chemicals (1 mentions) Penicillin streptomycin solution, by Thermo Fisher Scientific (1 mentions) Uplc beh c18 column, by Waters Corporation (1 mentions) Powerplex 16 hs system, by Promega (1 mentions) Lookout mycoplasma pcr detection kit, by Merck Group (1 mentions) Nu7441, by Selleck Chemicals (1 mentions) Hplc 1260 series system, by Bruker (1 mentions) Ft nmr spectrometer, by Bruker (1 mentions) Apex 4 70 ev ft ms spectrometer, by Bruker (1 mentions) Microtof qiii mass spectrometer, by Agilent Technologies (1 mentions) Agilent eclipse xdb c18 column, by Agilent Technologies (1 mentions) Acquity uplc amide column, by Waters Corporation (1 mentions)

8 protocols using uplc ms system

Amino Acid Analysis by HPLC-Fluorescence

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The analyses of amino acids were
carried out by the post column derivatization method with Fmoc-Cl.^{33 (link)} The samples containing hydroxyproline were analyzed
by using the Waters 2695 HPLC system equipped with a Diomansil C18
column (250 mm long with 4.6 mm in inner diameter). The chromatographic
conditions were as follows: mobile phase A [NaAc–HAc buffer
(50 mM, pH 4.2):acetonitrile, 70:30] and mobile phase B [NaAc–HAc
buffer (50 mM, pH 4.2):acetonitrile, 30:70] were used with a gradient
elution program at a flow rate of 1 mL·min^–1, and the column temperature was kept at 25 °C. The injection
volume was 10 μL. Fmoc-Cl derivatives of amino acids and hydroxyl
amino acids formed in the reaction mixture were detected spectrofluorometrically
at 263 nm.
LC–MS analysis was carried out using a Waters
ACQUITY UPLC–MS system with a Waters ACQUITY UPLC HSS C18 reversed
phase column (inner diameter: 1.8 μm). The inlet, MS transfer
line, and ion source temperatures were set at 280, 280, and 230 °C,
respectively.

Jing X., Wang X., Zhang W., An J., Luo P., Nie Y, & Xu Y. (2019). Highly Regioselective and Stereoselective Hydroxylation of Free Amino Acids by a 2-Oxoglutarate-Dependent Dioxygenase from Kutzneria albida. ACS Omega, 4(5), 8350-8358.

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Quantifying Cell Wall Precursor Accumulation in MRSA

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The intracellular accumulation of the cell wall precursor, UDP-MurNAc-pentapeptide after treatment of MRSA USA300 with malacidin was assessed as previous described.^{16 (link)} In brief, single colonies of MRSA USA300 were grown in LB media with and without 15 mM CaCl₂ to an OD₆₀₀ of 0.6. 1 mL of cells and media were incubated with a final concentration of 130 μg mL⁻¹ chloramphenicol for 15 min at 37 °C. Antibiotics to be assayed were added at 10 μg mL⁻¹ and incubated for 60 min at 37 °C. Vancomycin, known to form a complex with lipid II, was used as positive control. Cells were collected by centrifugation, and resuspended in 30 uL dH₂O and incubated in boiling water for 15 minutes. The cell extract was then centrifuged at 14,000 × g. Supernatant was analyzed for UDP-linked cell wall precursors by UPLC-MS system (Waters Corporation). Experiments were performed with biological replicates.

Hover B.M., Kim S.H., Katz M., Charlop-Powers Z., Owen J.G., Ternei M.A., Maniko J., Estrela A., Molina H., Park S., Perlin D.S, & Brady S.F. (2018). Culture-independent discovery of the malacidins as calcium-dependent antibiotics with activity against multidrug-resistant Gram-positive pathogens. Nature microbiology, 3(4), 415-422.

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Validating HepG2 Cell Cultures for Mycoplasma-Free Experiments

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HepG2 (ATCC, HB-8065) cells were cultured in 1× DMEM (Fisher Scientific, Cat#MT10013CM) supplemented with 10% FBS (Gibco, Cat#10437028), 1% Penicillin-Streptomycin solution (Gibco, Cat#15140122)) at 37 °C in 5% CO₂ in a humidified incubator. All human-derived cell lines were validated by short tandem repeat (STR) profiling using PowerPlex® 16 HS System (Promega) once a month. Additionally, a polymerase chain reaction (PCR)-based method was used to screen for mycoplasma once a month employing the LookOut® Mycoplasma PCR Detection Kit (MP0035, Sigma-Aldrich) and JumpStart™ Taq DNA Polymerase (D9307, Sigma-Aldrich) to ensure cells were free of mycoplasma contamination.
Doxorubicin, NU7441, Samuraciclib (ICEC0942), and EPZ015666 (GSK3235025) were purchased from Selleckchem. All other compounds used for screening were obtained from St Jude compounds deposit. AZD7648, Lot01, was purchased from Chemietek, and the quality was verified by Chemieteck by HPLC-MS and NMR, with purity >99.5%. The purity of AZD7648 was further verified in house by using Waters UPLC-MS system (Acquity PDA detector, SQ detector and UPLC BEH-C18 column). The mass spectrometer was acquired using MassLynx v. 4.1. The chromatographic conditions are as follows: flow rate: 1.0 mL/min, sample injection volume: 2 µL, column temperature: 55 °C, mobile phase: 0.1% formic acid in CH₃CN and H₂O.

Fang J., Singh S., Cheng C., Natarajan S., Sheppard H., Abu-Zaid A., Durbin A.D., Lee H.W., Wu Q., Steele J., Connelly J.P., Jin H., Chen W., Fan Y., Pruett-Miller S.M., Rehg J.E., Koo S.C., Santiago T., Emmons J., Cairo S., Wang R., Glazer E.S., Murphy A.J., Chen T., Davidoff A.M., Armengol C., Easton J., Chen X, & Yang J. (2023). Genome-wide mapping of cancer dependency genes and genetic modifiers of chemotherapy in high-risk hepatoblastoma. Nature Communications, 14, 4003.

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LC-MS Analysis of Enzymatic Reactions

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LC–MS analysis was performed using a UPLC-MS system (Waters, Milford, MA). To a PBS solution of 30 mM QA-2OMeSiR was added 274 nM DPP4 (recombinant dipeptidyl peptidase IV human, Catalog number: D4943-1VL, Lot: SLCJ0390, Sigma-Aldrich) or 2 nM PSA (recombinant NPEPPS, Catalog number: 6410-ZN, Lot: SHF0418101, R&D Systems), and the solutions were incubated at 37 °C for the indicated time. The enzyme reaction was quenched by two-fold dilution with MeOH. Aliquots of 10 mL of quenched samples were subjected to UPLC-MS, and the mass and absorbance spectra at 550 nm were analyzed. Gradient elution, A/B = 95/5 to 5/95 (0–3.5 min), 5/95 to 95/5 (4.0–4.1 min), 95/5 (4.1–5.0 min) (eluent A: H₂O containing 0.1% formic acid, eluent B: acetonitrile containing 0.1% formic acid). For QA-2OMeSiR, m/z = 558.25; for A-2OMeSiR, m/z = 430.19; for 2OMeSiR, m/z = 359.16.

Kawashima S., Yoshida D., Yoshioka T., Ogasawara A., Fujita K., Yanagiya M., Nagano M., Konoeda C., Hino H., Kitano K., Sato M., Hino R., Kojima R., Komatsu T., Kamiya M., Urano Y, & Nakajima J. (2022). Rapid imaging of lung cancer using a red fluorescent probe to detect dipeptidyl peptidase 4 and puromycin-sensitive aminopeptidase activities. Scientific Reports, 12, 9100.

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Metabolite Profiling of A. nidulans

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The transformant of A. nidulans was grown on solid CD-ST for 3 days and extracted with ethyl acetate. The organic phases were evaporated to dryness and dissolved in methanol for LC-MS analyses. LC-MS metabolite profiles were performed on a Waters UPLC-MS system with the following method: chromatographic separation was achieved with a linear gradient of 5–99% MeCN-H₂O (both with 0.02% v/v formic acid) in 10 min followed by 99% MeCN for 3 min and then 5% MeCN-H₂O for 3 min, with a flow rate of 0.4 mL/min. The MS data were collected in the m/z range 50–1500 in positive mode simultaneously.

Zhang J.M., Liu X., Wei Q., Ma C., Li D, & Zou Y. (2022). Berberine bridge enzyme-like oxidase-catalysed double bond isomerization acts as the pathway switch in cytochalasin synthesis. Nature Communications, 13, 225.

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Spectroscopic Characterization of Compounds

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UV spectra were detected on an Alltech UVIS-200 detector. IR spectra were determined on a Thermo Nicolet Nexus Is50 FT-IR spectrometer. The NMR spectra were acquired on a 400, 500, or 600 MHz Bruker FTNMR spectrometer. Chemical shifts are referenced to the solvent peaks at δ_H 2.50 and δ_C 39.52 for DMSO-d₆. Mass spectra were obtained from a Bruker APEX IV 70 eV FT-MS spectrometer. The chromatographic (CC) substrates included silica gel (100−200 and 200−300 mesh) and HF254 silica gel for thin-layer chromatography (TLC) (Qingdao Marine Chemistry Co., Ltd., Qingdao, China). High-performance liquid chromatography (HPLC) was performed with an Alltech 426 pump equipped with an Alltech UVIS-200 detector (210 nm) and using a semipreparative reversed-phase column (YMC-packed, C₁₈, 5 μM, 10 × 250 mm). UPLC was performed on an Agilent UPLC series 1200 (Agilent Technologies, Santa Clara, CA, USA) equipped with an Agilent Eclipse XDB-C18 column (5 μm, 4.6 × 150 mm). LC-MS analysis was carried out on an Agilent HPLC 1260 series system equipped with a Bruker microTOF QIII mass spectrometer by using an Agilent Eclipse XDB C18 column (5 μm, 4.6 × 150 mm) or Waters UPLC-MS system. The chemicals used in this study were obtained from Beijing Tongguang Fine Chemicals Company of the highest available purity.

Huang Z., Liu D., Chen S., Ren J., Gao C., Li Z., Fan A, & Lin W. (2024). Brominated Depsidones with Antibacterial Effects from a Deep-Sea-Derived Fungus Spiromastix sp.. Marine Drugs, 22(2), 78.

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Quantifying Cell Wall Precursor Accumulation in MRSA

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UPLC-MS Analysis of Plant Sugars

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To analyze the sugar components in plants, the plant juice was filtered, was mixed with methanol (1:1, vol/vol), and was loaded onto a Waters UPLC-MS system (Waters).
An Acquity UPLC Amide column (2.1 × 100 mm, Waters) was employed for chromatography. Solution A was composed of 0.1% NH 4 OH in water, and solution B was composed of 0.01% formic acid in acetonitrile. An isocratic elution of 75% B was performed for 10 min. The entire column eluate was introduced into a Q-TOF mass spectrometer. Ion detection was conducted under the same conditions as described above.

Zhang C., Lv M., Yin W., Dong T., Chang C., Miao Y., Jia Y, & Deng Y. (2019). Xanthomonas campestris Promotes Diffusible Signal Factor Biosynthesis and Pathogenicity by Utilizing Glucose and Sucrose from Host Plants. Molecular plant-microbe interactions : MPMI, 32(2).

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