Accucore

Manufactured by Thermo Fisher Scientific

Sourced in Germany

Accucore is a high-performance liquid chromatography (HPLC) column developed by Thermo Fisher Scientific. It is designed to provide efficient and reliable separation of a wide range of analytes. The Accucore column utilizes core-shell particle technology to deliver high-resolution chromatography, reduced analysis time, and improved peak shape.

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9 protocols using accucore

Quantitative Proteomic Analysis Pipeline

Cited 2 times

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Labeled peptides were resuspended in 5% ACN/2% formic acid at 1 mg/mL and loaded at 1 μg, unless otherwise noted, on an in-house pulled C₁₈ column (30 cm, 2.6 um Accucore [Thermo Fisher], 100um ID), and eluted using a linear gradient from 0% to 30% buffer B (95% ACN, 0.125% formic acid). Eluted peptides were injected into an Orbitrap Fusion Lumos (Tune 3.1.2412) using a either a high-resolution MS² (HRMS²) or synchronous precursor selection (SPS-MS³) method for quantitation (Supplementary Methods). All figure panels include replicate injections separated on the same analytical column. Acquired Raw files were searched with SEQUEST^{15 (link)} using an in house proteomic pipeline (Supplementary Methods). Peptide spectral matches were first filtered to a peptide false discovery rate of less than 1% based on linear discriminant analysis using a target decoy strategy.^{16 (link),17 (link)} Peptides were subsequently filtered to a final protein-level false discovery rate less than 1%.^{16 (link),18 (link)} For quantitation, a total sum signal-to-noise of all reporter ions of 200 was required for analysis and comparisons. Data analysis was performed using the R statistical scripting language (3.5.1, “Feather Spray”).¹⁹

Schweppe D.K., Prasad S., Belford M.W., Navarrete-Perea J., Bailey D.J., Huguet R., Jedrychowski M.P., Rad R., McAlister G., Abbatiello S.E., Woulters E.R., Zabrouskov V., Dunyach J.J., Paulo J.A, & Gygi S.P. (2019). Characterization and Optimization of Multiplexed Quantitative Analyses Using High-Field Asymmetric-Waveform Ion Mobility Mass Spectrometry. Analytical chemistry, 91(6), 4010-4016.

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HPLC Analysis of Radioactive Compounds

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HPLC analysis was performed using an Ultimate 3000 system (Thermo Fisher Scientific) with a diode-array UV detector and a Geiger–Müller tube-based radiodetector (Scansys Laboratorieteknik, Denmark). A C18 reversed-phase column (Accucore, 2.6 µm, C18, 100 Å, 150 × 4.6 mm, Thermo Fisher Scientific) was eluted with 75% acetonitrile in 25 mM sodium phosphate buffer (pH 6.2); injection volume, 50 µL; flow rate, 1.5 mL/min; on-line UV (220 nm) and radioactivity detection.

Bratteby K., Denholt C.L., Lehel S., Petersen I.N., Madsen J., Erlandsson M., Ohlsson T., Herth M.M, & Gillings N. (2021). Fully Automated GMP-Compliant Synthesis of [18F]FE-PE2I. Pharmaceuticals, 14(7), 601.

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UHPLC-PDA-Orbitrap Metabolites Characterization

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Liquid chromatography was performed using an UHPLC C18 column (Accucore, 150 mm × 4.6 mm ID, 2.5 μm, Thermo Fisher Scientific, Bremen, Germany) operated at 25°C. The detection wavelengths were 254, 280, 330 and 354 nm, and PDA was recorded from 200 to 800 nm for peak characterization. Mobile phases were 1% formic aqueous solution (A) and 1% formic acid in acetonitrile (B). The gradient program (time (min), % B) was: (0.00, 12); (5.00, 12); (10.00, 20); (15.00, 40); (20.00, 40); (25.00, 70); (35.00, 12); and 15 min for column equilibration before each injection. The flow rate was 1.00 mL min⁻¹, and the injection volume was 10 μL. The standards, and the extracts dissolved in ethanol, were kept at 10°C during storage in the autosampler. The HESI II and Orbitrap spectrometer parameters were optimized as previously reported (Salgado et al., 2017 (link); Torres-Benitez et al., 2017 (link)).

Calla-Quispe E., Robles J., Areche C, & Sepulveda B. (2020). Are Ionic Liquids Better Extracting Agents Than Toxic Volatile Organic Solvents? A Combination of Ionic Liquids, Microwave and LC/MS/MS, Applied to the Lichen Stereocaulon glareosum. Frontiers in Chemistry, 8, 450.

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Chromatographic Analysis of Resin Extracts

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Liquid chromatography was performed using an UHPLC C18 column (Accucore, 150 mm × 4.6 mm internal diameter, 2.5 μm particle size, Thermo Fisher Scientific, Bremen, Germany) operated at 25 °C. The detection wavelengths were 254, 280, 330 and 354 nm, and PDA was recorded from 200 to 800 nm for peak characterization. Mobile phases were 1% formic aqueous solution (A) and 1% formic acid in acetonitrile (B). The gradient program (time (min), % B) was: (0.00, 12); (5.00, 12); (10.00, 20); (15.00, 40); (20.00, 40); (25.00, 70); (35.00, 12) and 15 min for column equilibration before each injection. The flow rate was 1.00 mL min⁻¹, and the injection volume was 10 µL. Standards and the resin extract dissolved in methanol were kept at 10 °C during storage in the autosampler. The HESI II and Orbitrap spectrometer parameters were optimized as previously reported [24 (link),42 (link)].

Ardiles A., Barrientos R., Simirgiotis M.J., Bórquez J., Sepúlveda B, & Areche C. (2018). Gastroprotective Activity of Parastrephia quadrangularis (Meyen), Cabrera from the Atacama Desert. Molecules : A Journal of Synthetic Chemistry and Natural Product Chemistry, 23(9), 2361.

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Purification and Characterization of Organic Compounds

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THF, DCM, toluene, and acetonitrile were obtained via a solvent purification system by filtering through two columns packed with activated alumina and 4 Å molecular sieve, respectively. All other chemicals obtained from commercial sources were used without further purification. Flash chromatography was performed using silica gel (230–400 mesh) as the stationary phase. Reaction progress was monitored by thin-layer chromatography (silica-coated glass plates) and visualized by UV light, and/or by LC-MS. NMR spectra were recorded in CDCl_3, CD₃OD, or acetone-d6 at 400 or 600 MHz for ¹H NMR. Chemical shifts δ are given in using tetramethylsilane as an internal standard. Multiplicities of NMR signals are designated as singlet (s), broad singlet (br s), doublet (d), doublet of doublets (dd), triplet (t), quartet (q), and multiplet (m). All final compounds for biological testing were of ≥95.0% purity as analyzed by LC-MS, performed on an Advion AVANT LC system with the expression CMS using two different columns: a Thermo Accucore™ Vanquish™ C18+ UHPLC Column (1.5 μm, 50 × 2.1 mm) at 40 °C and a Thermo Scientific™ BetaSil™ C18 Column (3.0 μm, 150 × 4.6 mm) at 25 °C. Gradient elution was used for UHPLC with a mobile phase of acetonitrile and water containing 0.1% formic acid.

Zhang X., Thummuri D., Liu X., Hu W., Zhang P., Khan S., Yuan Y., Zhou D, & Zheng G. (2020). Discovery of PROTAC BCL-XL Degraders as Potent Anticancer Agents with Low On-target Platelet Toxicity. European journal of medicinal chemistry, 192, 112186.

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Orbitrap Fusion Peptide Analysis

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Labeled peptides were resuspended in 5% ACN/5% formic acid at 1 mg/mL and loaded at 1 μg, unless noted otherwise, on an in-house pulled C18 column (30−35 cm, 2.6 μm Accucore (Thermo Fisher), 100 μm ID), and eluted using a linear gradient from 100% buffer A (5% ACN, 0.125% formic acid) to 30% buffer B (95% ACN, 0.125% formic acid) using an Easy-nLC 1000 (Thermo Scientific). Eluted peptides were analyzed by an Orbitrap Fusion or Fusion Lumos mass spectrometer over a 1–3 h gradient. Instrument parameter details, corresponding to a given figure, are outlined in Table S1. In all cases, the dispersion voltage is set at −5000 V. MS1 features were determined using RawFileReader (https://planetorbitrap.com/rawfilereader).

Schweppe D.K., Rusin S.F., Gygi S.P, & Paulo J.A. (2019). Optimized Workflow for Multiplexed Phosphorylation Analysis of TMT-Labeled Peptides Using High-Field Asymmetric Waveform Ion Mobility Spectrometry. Journal of proteome research, 19(1), 554-560.

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HPLC-MS Analysis of Bioactive Compounds

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HPLC was performed using a C18 column (Accucore, 150 mm × 4.6 mm ID, 2.5 µm, Thermo Fisher Scientific, Bremen, Germany) at 25 °C. Detection wavelengths were 254, 280, 330, and 354 nm, and PDA from 200 to 800 nm for peak identification. The mobile phase was 0.1% formic acid aqueous solution (A) and 0.1% formic acid in acetonitrile (B). The gradient program (time (min), % B) was: (0.00, 12); (5.00, 12); (10.00, 20); (15.00, 40); (20.00, 40); (25.00, 70); (35.00, 12) and 15 min for column equilibration before each injection. The flow rate was 1.00 mL min⁻¹, and the injection volume was 10 µL. The extracts and standards dissolved in methanol were kept at 10 °C during storage in the auto sampler. The parameters of the Impact HD mass spectrometer were set as previously described [8 (link),34 (link),36 (link)].

Guillen E., Terrones H., de Terrones T.C., Simirgiotis M.J., Hájek J., Cheel J., Sepulveda B, & Areche C. (2024). Microwave-Assisted Extraction of Secondary Metabolites Using Ethyl Lactate Green Solvent from Ambrosia arborescens: LC/ESI-MS/MS and Antioxidant Activity. Plants, 13(9), 1213.

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Lipidomic analysis of liver and plasma

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Liver and plasma samples were extracted using the Bligh‐Dyer extraction method.⁽⁹⁾ Liquid chromatography was performed on an ultra‐high‐performance liquid chromatography (UHPLC) system (Vanquish; Thermo Scientific, San Jose, CA) using a C₃₀ column (2.1 × 150 mm, 2.6 μm particle size) (Accucore; Thermo Scientific) and a gradient program consisting of mobile phase A (60:40 acetonitrile/water) and mobile phase B (90:10 isopropanol/acetonitrile), each containing 5 mmol/L ammonium formate and 0.1% (vol/vol) formic acid. The UHPLC system was coupled to a mass spectrometer (Q‐Exactive Orbitrap; Thermo Scientific) for chromatographic separation and mass spectral measurement of lipids in positive and negative ion mode, respectively, using full‐scan and iterative exclusion⁽¹⁰, ¹¹⁾ tandem mass spectrometry. The software LipidMatch Flow⁽¹²⁾ was deployed for peak picking, blank filtering, annotation, and combining positive and negative ion polarity, whereas LipidMatch Normalizer⁽¹³⁾ was used for semiquantitation. Novel oxidized lipidomics libraries were developed and applied as reported.⁽¹⁴⁾ Extensive details on software development, deployment, and acquisition methods can be found in the Supporting Material.

Koelmel J.P., Tan W.Y., Li Y., Bowden J.A., Ahmadireskety A., Patt A.C., Orlicky D.J., Mathé E., Kroeger N.M., Thompson D.C., Cochran J.A., Golla J.P., Kandyliari A., Chen Y., Charkoftaki G., Guingab‐Cagmat J.D., Tsugawa H., Arora A., Veselkov K., Kato S., Otoki Y., Nakagawa K., Yost R.A., Garrett T.J, & Vasiliou V. (2021). Lipidomics and Redox Lipidomics Indicate Early Stage Alcohol‐Induced Liver Damage. Hepatology Communications, 6(3), 513-525.

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LC-MS Metabolite Profiling of Urine and Plasma

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Urine and plasma samples were analysed separately in randomised batches. In brief, LC-MS analyses were carried out on a Dionex UltiMate 3000 RSLC HPLC System (Dionex), equipped with an UltiMate 3000 RS pump, an UltiMate 3000 RS autosampler and a QExactive Quadrupole-Orbitrap Mass Spectrometer (Thermo Fisher Scientific). Electrospray ionisation at both negative and positive ion modes was performed with a spray voltage of 2•00 kV and capillary temperature of 280°C. The total ion current with a range of 50-1000 m/z and 70 000 resolution was measured. Sample aliquots (2 µl) were injected on an Accucore (Thermo Fisher) C18 (150 × 2•5 mm, 5-μm particle size) reverse-phase column, thermostatically regulated at 40°C. The mobile phase consisted of water with 0•1 % formic acid (solvent A) and acetonitrile with 0•1 % formic acid (solvent B). Starting conditions were 95 % A, and compounds were separated with a gradient of 95 % A to 5 % A over 18 min, with a 5-min wash cycle before being re-equilibrated to 95 % A; flow rate was 0•35 ml/min. All samples were randomised in their running order, and a quality-control sample was analysed on ten occasions throughout the batch. The quality-control was a pool of all samples in a single aliquot.

Langer S., Kennel A, & Lodge J.K. (2018). The influence of juicing on the appearance of blueberry metabolites 2 h after consumption: a metabolite profiling approach. The British journal of nutrition, 119(11).

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