Ascentis express rp amide column

Manufactured by Merck Group

The Ascentis Express RP-Amide column is a reversed-phase high-performance liquid chromatography (HPLC) column. It features a silica-based stationary phase with amide functionality, which allows for the separation of a wide range of analytes, including polar and moderately polar compounds.

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Lab products found in correlation

Ionpac as11 column, by Thermo Fisher Scientific (1 mentions) Rf 20a xs detector, by Shimadzu (1 mentions) Nexera uplc system, by Shimadzu (1 mentions) Lcms 8040 system, by Shimadzu (1 mentions) Nexera x2 system, by Shimadzu (1 mentions) Waters acquity uplc, by Waters Corporation (1 mentions) Xevo tq triple quadrupole mass spectrometer, by Waters Corporation (1 mentions)

Close spelling variants of this product

Ascentis Express RP-amide C16 column Ascentis® Express 2.7 μ RP-Amide column Ascentis Express RP-Amide analytical column Ascentis Express RP-Amide Ascentis RP-Amide column Ascentis RP-Amide RP-amide column Ascentis Express

6 protocols using ascentis express rp amide column

Analytical RP-Amide Chromatography of RA Isomers

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Resolution of RA isomers was achieved with an analytical Ascentis Express RP-Amide column (100 × 2.1 mm, 2.7 μm, Sigma-Aldrich, 53913-U) fitted with a preceding guard column at a flow rate of 500 μl/min. The following were the mobile phases used: (A) 0.1% formic acid in water/methanol (6:4); (B) 0.1% formic acid in acetonitrile/methanol (6:4). A gradient was applied over 25 min: 0 to 2 min, 40% B; 2 to 10 min, 40 to 55% B; 10 to 18 min, 55 to 95%; 18 to 21 min, holding at 95% B; 21 to 23 min, 95 to 40% B; 23 to 25 min, holding at 40% B.

Yoo H.S., Moss K.O., Cockrum M.A., Woo W, & Napoli J.L. (2023). Energy status regulates levels of the RAR/RXR ligand 9-cis-retinoic acid in mammalian tissues: Glucose reduces its synthesis in β-cells. The Journal of Biological Chemistry, 299(10), 105255.

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Hydrophilic Interaction Anion Exchange HPLC

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Hydrophilic interaction anion exchange (HIAX) HPLC was performed with an IonPac AS11 column (Dionex, Sunnyvale, USA; 4 × 250 mm, combined with a 4 × 50mm guard column) on a Shimadzu Nexera UPLC system as described previously 12 (link),13 (link). A two solvent gradient with buffer A (0.8 M ammonium acetate, pH 3) and buffer B (80% acetonitrile; LC-MS grade) was applied at a flow rate of 1 ml/min: 0-5 min, 99% B; 5-50 min, 90% B; 50-65 min, 80% B; 65-85 min, 75% B. A pool of pyridylaminated (PA) oligomannosidic N-glycans from white beans (containing Man_3-9GlcNAc₂) was used to calibrate the column and test efficiency of separation. For RP-HPLC isomeric/isobaric comparison with wild-type glycans, an Ascentis Express RP-amide column (150 x 4.6 mm, 2.7 µm; Sigma Aldrich, calibrated in terms of glucose units with an oligoglucose standard) was employed with a gradient of 30% (v/v) methanol (buffer B) in 100 mM ammonium acetate, pH 4 (buffer A) being applied at a flow rate of 0.8 ml/min as follows: 0-4 min, 0% B; 4-14 min, 0-5% B; 14-24 min, 5-15% B; 24-34 min, 15-35% B; 34-35 min, return to starting conditions 20 (link). The fluorescence was measured at 320 nm (excitation) and 400 nm (emission) with a Shimadzu RF-20A XS detector.

Hykollari A., Malzl D., Yan S., Wilson I.B, & Paschinger K. (2017). Hydrophilic Interaction Anion Exchange for Separation of Multiply-Modified Neutral and Anionic Dictyostelium N-Glycans. Electrophoresis, 38(17), 2175-2183.

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Quantification of Hepatic all-trans Retinoic Acid

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Hepatic atRA levels were measured by LC‐MS/MS, as previously described.23 Briefly, liver tissues (~ 100 mg) were homogenized in a 1:1 volume of 0.9% NaCl, and the sample was transferred into a 15‐mL glass culture tube. Two volumes of acetonitrile containing 0.1% formic acid were added along with D5‐atRA (internal standard). Retinoic acid was extracted using 10 mL of hexanes, and the organic layer was dried under nitrogen at 37°C. The samples were reconstituted in 65 μL of acetonitrile/H₂O (60:40) for LC‐MS/MS analysis. Standard curve and quality control samples were generated using ultraviolet (UV) light–exposed mouse liver homogenate spiked with atRA at concentrations of 0, 2, 5, 10, 15, and 20 nM for the standards and 3, 7.5, and 17.5 nM for the quality control samples. The retinoids were separated using an Agilent 1290 UPLC (Santa Clara, CA) equipped with a Sigma (St. Louis, MO) Ascentis Express RP Amide column (2.7 μm; 150 mm × 2.1 mm). Analytes were detected using an AB Sciex 5500 QTRAP MS/MS (Foster City, CA) operated in positive ion atmospheric pressure chemical ionization mode. MRM transitions for atRA and internal standard were 301/205 and 306/116, respectively.

Ning M., Duarte J.D., Stevison F., Isoherranen N., Rubin L.H, & Jeong H. (2019). Determinants of Cytochrome P450 2D6 mRNA Levels in Healthy Human Liver Tissue. Clinical and Translational Science, 12(4), 416-423.

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Simultaneous Quantification of Steroid Hormones

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DHEA, E2, and T were separated via reverse-phase chromatography with an Agilent 1290 system (Santa Clara, CA, U.S.A.) equipped with a binary pump, column compartment, and autosampler. The column compartment was maintained at 40 °C: samples were kept in the autosampler at 10 °C. Separation was achieved with an analytical Ascentis Express RP-Amide column (100 × 2.1 mm, 2.7 μm, Sigma Aldrich) at a flow rate of 0.4 mL/min. Mobile phases were (A) 0.1% formic acid in water; (B) 0.1% formic acid in methanol. The following gradient was applied over a run time of 25 min: 0 to 2 min, 50% B; 2 to 8 min, 50 to 95% B; 8 to 20 min, holding at 95% B; 20 to 23.5 min, 95 to 50% B; 23.5 to 25 min, back to 50% B and re-equilibrating for 1.5 min.

Yoo H.S, & Napoli J.L. (2019). Quantification of Dehydroepiandrosterone, 17β-Estradiol, Testosterone, and Their Sulfates in Mouse Tissues by LC-MS/MS. Analytical chemistry, 91(22), 14624-14630.

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Quantification of Chlorogenic Acid Derivatives

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Onopordum extracts were analysed using a Shimadzu Nexera X2 system equipped with a photodiode detector SPD-M20A in series to a triple quadrupole Shimadzu LCMS-8040 system provided with electrospray ionization (ESI) source (Shimadzu, Düsseldorf Germany). An Ascentis Express RP-Amide column (10cmx2.1mmx2.7µm, Supelco, Chlorogenic acid and 1,3 dicaffeoylquinic acid were used for the quantification of chlorogenic acid derivatives and the dicaffeoylquinic and succinyl dicaffeoylquinic acids, respectively. Each standard solution and extracts were analysed in two replicates.
Calibration curves were prepared with five different concentrations, in the range of 0.1-5 µg/mL, monitoring the reported transitions: ESI + : m/z 355.00 →163.00, for chlorogenic acid and 517.00 →163.00 for 1,3 dicaffeoylquinic acid. (dwell time: 20 msec, collision energy -35 V, event time: 0.096 sec). The determination coefficients were 0.993 and 0.995 for chlorogenic acid and 1,3 dicaffeoylquinic acid, respectively.

Marengo A., Fumagalli M., Sanna C., Maxia A., Piazza S., Cagliero C., Rubiolo P., Sangiovanni E, & Dell'Agli M. (2018). The hydro-alcoholic extracts of Sardinian wild thistles (Onopordum spp.) inhibit TNFα-induced IL-8 secretion and NF-κB pathway in human gastric epithelial AGS cells. Journal of ethnopharmacology, 210.

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Quantification of FPR Agonists in Mouse Plasma

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Plasma samples collected from mouse Cohorts 1 and 3 (after 24 and 48 h reperfusion, respectively), ∼20 h post-final dose, were analysed for plasma FPR agonist concentrations by the Monash University Centre for Drug Candidate Optimisation (Parkville, VIC, Australia). The FPR agonists were extracted from plasma samples using acetonitrile precipitation (2:1) and analysis was conducted using a Waters Xevo TQ triple quadrupole mass spectrometer coupled to a Waters Acquity UPLC (Waters Corporation, Milford, MA). Mass spectrometry was performed in positive-mode electrospray ionization using multiple reaction monitoring, with MS–MS transitions of 456.01>285.07 and 385.11>231.96 for Cmpd17b and Cmpd43, respectively. UPLC conditions comprised a Supelco Ascentis Express RP Amide column (50 × 2.1 mm, 2.7 μm), with an acetonitrile–water–0.05% formic acid gradient mobile phase, gradient cycle time of 4 min and flow rate 0.4 ml min⁻¹. The elution times of Cmpd17b and Cmpd43 were 2.24 and 1.66 min, respectively, while that of the internal standard diazepam was 1.79 min. The lower limit of detection for both FPR agonists was 1 ng ml⁻¹.

Qin C.X., May L.T., Li R., Cao N., Rosli S., Deo M., Alexander A.E., Horlock D., Bourke J.E., Yang Y.H., Stewart A.G., Kaye D.M., Du X.J., Sexton P.M., Christopoulos A., Gao X.M, & Ritchie R.H. (2017). Small-molecule-biased formyl peptide receptor agonist compound 17b protects against myocardial ischaemia-reperfusion injury in mice. Nature Communications, 8, 14232.

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