Gemini c18 reverse phase column

Manufactured by Phenomenex

Sourced in Italy

The Gemini C18 reverse phase column is a chromatographic separation tool designed for the analysis of a wide range of organic compounds. It features a spherical silica-based stationary phase with a chemically bonded octadecyl (C18) ligand. The column is suitable for the separation and purification of various analytes, including pharmaceuticals, natural products, and environmental contaminants.

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Lab products found in correlation

Pre column, by Phenomenex (1 mentions) Facs caliber, by BD (1 mentions) Lachrom elite hplc system, by Hitachi (1 mentions) Diode array detector, by Agilent Technologies (1 mentions) Securityguard c18, by Phenomenex (1 mentions) Agilent 1100 series hplc system, by Agilent Technologies (1 mentions) Lc 2050c 3d system, by Shimadzu (1 mentions) Ultrafiltration units, by Merck Group (1 mentions) Ultimate 3000 rslcnano hplc system, by Thermo Fisher Scientific (1 mentions) Kinetex c18 column, by Phenomenex (1 mentions) Totalchrom software, by PerkinElmer (1 mentions)

Close spelling variants of this product

Gemini C18 reverse-phase Gemini C18 column Gemini C18 Gemini column C18 column

8 protocols using gemini c18 reverse phase column

HPLC Analysis of Olive Leaf Extract

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A high-performance liquid chromatography analysis (HPLC) was carried out to identify and quantify the major phenolic compounds of the OLE using a slightly modified method developed in our previous study [40 (link)].
The retention times and UV absorbance spectra of phenolic compounds present in OLE were compared with those of the commercial standard and quantified at 278 nm, using p-hydroxyphenyl acetic acid as the internal standard, according to the previously reported method. Sample concentrations were determined by linear regression. For each calibration curve, the correlation coefficients were >0.99.
HPLC analysis was performed using an HPLC instrument (Beckman, Ramsey, MN) equipped with a System Gold Solvent Delivery module (Pumps) 125 and a System Gold UV/Vis Detector 166, set to 280 nm, and using a Phenomenex Gemini reverse-phase C18 column (250 × 4.6 mm, 5 μm particle size; Phenomenex, Castel Maggiore, Italy). The mobile phase was a mixture of H₂O/AcOH (97.5:2.5 v/v) (A) and MeOH/ACN (1:1 v/v) (B), programmed as follows: a gradient from 5% (B) to 30% (B) in 45 min; 30% (B) for 5 min and then from 30% (B) to 5% (B) in 5 min. The flow rate was 1 mL/min, and the injected volume was 50.0 μL.

De la Ossa J.G., Felice F., Azimi B., Esposito Salsano J., Digiacomo M., Macchia M., Danti S, & Di Stefano R. (2019). Waste Autochthonous Tuscan Olive Leaves (Olea europaea var. Olivastra seggianese) as Antioxidant Source for Biomedicine. International Journal of Molecular Sciences, 20(23), 5918.

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Metabolite Separation by HILIC-LC-MS/MS

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Resuspended metabolites were separated by HILIC with a Gemini reverse-phase C18 column (50 mm × 4.6 mm with 5 μm diameter particles) from Phenomenex together with precolumn (C18, 3.5 mm, 2 mm × 20 mm). Mobile phase A was composed of 100% H₂O (10 mmol/L tributylamine aqueous solution, adjusted to pH 4.95 with 15 mmol/L acetic acid), and mobile phase B was composed of 100% methanol. We used a multistep gradient with buffer A and B as follows: from 0 to 5 minutes, 95% buffer A; from 5 to 15 minutes, linear change from 95% to 90% buffer A; from 15 to 22 minutes, linear change from 90% to 85% buffer A; from 22 to 26 minutes, 10% buffer A; and from 30 to 33 minutes, 95% buffer A, and maintained for 7 minutes. The flow rate was 0.2 mL/minute for 0 to 15 minutes and 30 to 40 minutes, and 0.3 mL/minute for 15 to 30 minutes. Targeted MS/MS analysis was performed on an Agilent triple quadrupole LC-MS/MS instrument (Agilent Technologies 6460 QQQ). The capillary voltage was set to 4.0 kV. The drying gas temperature was 350°C, the drying gas flow rate was 10 L/minute, and the nebulizer pressure was 45 psi. Relative metabolite abundance was quantified by integrated peak area for the given MRM transition. Data presented are representative of 3 independent biological experiments each containing 3 technical replicates for a given condition.

Stein B.D., Ferrarone J.R., Gardner E.E., Chang J.W., Wu D., Hollstein P.E., Liang R.J., Yuan M., Chen Q., Coukos J.S., Sindelar M., Ngo B., Gross S.S., Shaw R.J., Zhang C., Asara J.M., Moellering R.E., Varmus H, & Cantley L.C. (2023). LKB1-Dependent Regulation of TPI1 Creates a Divergent Metabolic Liability between Human and Mouse Lung Adenocarcinoma. Cancer Discovery, 13(4), 1002-1025.

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Solvent Evaporation and Chromatographic Analysis Protocol

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Solvent evaporation was achieved under vacuum by using a rotating evaporator (Strike 300, Stereoglass, Perugia, Italy). The homogenization was achieved by using a vortex mixer (Zx3, Advanced Vortex Mixer, VELP^® Scientifica, Usmate Velate, Italy) and a rotary shaker. Centrifugation was performed by using a centrifuge 4225 at 4000 rpm.
HPLC analysis was performed with an HPLC instrument (Thermo Finnigan-Spectra System SCM1000, Thermo Electron Corporation, Waltham, MA, USA) equipped with a Spectra System P2000 (Thermo Electron Corporation, Waltham, MA, USA ), Spectra System UV2000, set to 280 nm, and by using a Phenomenex Gemini reverse-phase C18 column (250 × 4.6 mm, 5 μm particle size; Phenomenex, Castel Maggiore, Italy).
Spectrophotometric analysis was carried out with a spectrophotometer (Shimadzu, Columbia, MD, USA) set at 725 nm.

Esposito Salsano J., Digiacomo M., Cuffaro D., Bertini S, & Macchia M. (2022). Content Variations in Oleocanthalic Acid and Other Phenolic Compounds in Extra-Virgin Olive Oil during Storage. Foods, 11(9), 1354.

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Quantification of Sialic Acid Content

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Sera-Mag SpeedBeads Blocked Streptavidin (Thermo-Fisher cat# 2115-2104-011150, 1 µm diameter) were washed three times and resuspended in 125 mM PBS pH 7.4 (Gibco). Biotinylated mucin mimetic polymers were added at final concentration of 20–300 nM, and incubate 45 min at room temperature with rotation. The polymers-conjugated beads were washed with PBS pH 7.2 (Gibco) three times, and resuspended in PBS pH 7.2 supplemented with 0.02% sodium azide. Polymer conjugation to the beads was quantified by flow cytometry (BD FACSCaliber, BD-Biosciences). Sialic acid content of 6’sialyllactose, 3’sialyllactose and lactose beads was determined by 1,2-diamino-4,5-methylenedioxybenzene dihydrochloride–high performance liquid chromatography analysis (DMB-HPLC) according to a published protocol (35 (link)). Samples were incubated for 1h with 0.1M HCl at 80°C to release Sias, and filtered through microcon-3kDa filtration device (Millipore). Free Sialic acids were incubated for 2.5 h at 50°C in the dark with 7 mM DMB (Sigma), 0.75 M 2-mercapto-ethanol, 18 mM Na-hydrosulfite in 1.4 M acetic acid. Sialic acids were separated on 250x4.6 mm Gemini C18 reverse phase column (Phenomenex) with 7% MeOH, 8% Acetonitrile, 85% H₂O solution at 0.9 ml/min using the ELITE Lachrom HPLC system (Hitachi). DMB-labeled Sialic acids were detected at EX=373 EM=488.

Cohen M., Fisher C.J., Huang M.L., Lindsay L.L., Plancarte M., Boyce W.M., Godula K, & Gagneux P. (2016). Capture and characterization of influenza A virus from primary samples using glycan bead arrays. Virology, 493, 128-135.

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HPLC Analysis of Cx43MP Compound

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HPLC was performed on an Agilent 1100 series HPLC system coupled with a diode array detector (Agilent, USA). Chromatographic separation was achieved using a Gemini^® C₁₈ reverse phase column (150 mm × 3 mm, 3 µm 110Ǻ, Phenomenex; PN 00F-4439-Y0) protected with a SecurityGuard™ C₁₈ (8mm×3 mm, 3 µ, Phenomenex; PN AJ0-4287) guard column. Chromatographic separation was performed in gradient mode with a flow rate of 0.4 mL/min consisting of an aqueous (B, 0.01% (v/v) FA in Milli-Q water) and an organic phase (A, 0.01% (v/v) FA in ACN) with the column temperature set to 30 °C. The injection volume was 10 µL and UV detection of Cx43MP was accomplished at 214 nm. The volume of injection loop was 20 µL. The gradients used are 0 min (A-5% and B-95%), 15 min (A-50% and B-50%), 17 min (A-100% and B-0%), 19 min (A-5% and B-95%) and 25 min (A-5% and B-95%). Instrument control and data acquisition was achieved by ChemStation B.04.03-SP2 (Agilent, USA).

Bisht R., Rupenthal I.D., Sreebhavan S, & Jaiswal J.K. (2017). Development of a novel stability indicating RP-HPLC method for quantification of Connexin43 mimetic peptide and determination of its degradation kinetics in biological fluids. Journal of Pharmaceutical Analysis, 7(6), 365-373.

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Reverse-Phase HPLC for PLA Enantiomer Purity

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To evaluate the purity of the synthesized d- or l-PLA ILs and for the determination of their water solubility, a reverse phase-HPLC method was developed. The HPLC apparatus consisted of a Shimadzu LC2050C-3D system with a PDA detector. Samples were run through a C₁₈ precolumn and a Gemini C₁₈ reverse-phase column [150 × 4.5 mm (I.D.)] with 5 μm particle size packing (Phenomenex, Torrance, CA). The mobile phase contained acetonitrile and 20 mM phosphoric acid solution in water (25 : 75). The flow rate of the mobile phase was set at 0.8 mL min⁻¹, the column oven was set at 45 °C, the injection volume was 10 μL, and the detection was carried out at 210 nm. The retention time for PLA was 5.5 min. For the standard curve, d- or l-PLA stock solutions (0.1 mg mL⁻¹) were prepared in methanol. The stock solutions were diluted to obtain solutions of various concentrations. The standard curve was obtained by injecting 5–25 μg mL⁻¹ of PLA. All the experiments were performed in triplicate.

Crossley P., Sutar Y., Tsoy I., Mukkirwar S., Łaniewski P., Herbst-Kralovetz M.M, & Date A.A. (2024). Development of phenyllactic acid ionic liquids and evaluation of cytotoxicity to human cervical epithelial cells. RSC Advances, 14(23), 16083-16092.

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Encapsulation Efficiency of Oils in Nanoparticles

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The efficiency of encapsulation of the oils in the nanoparticles was evaluated by the ultrafiltration/centrifugation method, using Millipore ultrafiltration units with exclusion pore size of 10 kDa. The concentrations of unencapsulated CVC and LNL in the filtrate were determined by HPLC, and the encapsulation efficiency was calculated as the difference between the initial concentration added (100%) and the concentration in the ultrafiltrate. The analyses (in triplicate) were performed with an UltiMate 3000 RSLCnano HPLC system (Thermo Scientific). For CVC, a Phenomenex Gemini C18 reverse phase column (100 × 4.6 mm; 2.6 μm) was used, and the mobile phase consisted of acetonitrile:water (50:50, v/v), at a flow rate of 1 mL/min. For LNL, a Phenomenex Kinetex C18 column (250 × 4.6 mm; 3 μm) was employed, and the mobile phase consisted of acetonitrile:water (65:35, v/v), at a flow rate of 1.5 mL/min. The analytical curves used for quantification could be described by the following equations:

CVC = 5.4666 \times - 0.065 (r^{2} = 0.9989) and LNL = 1.2262 \times + 1.6471 (r^{2} = 0.9990) .

Campos E.V., Proença P.L., Oliveira J.L., Melville C.C., Della Vechia J.F., de Andrade D.J, & Fraceto L.F. (2018). Chitosan nanoparticles functionalized with β-cyclodextrin: a promising carrier for botanical pesticides. Scientific Reports, 8, 2067.

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Dacarbazine Photoproduct Separation

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Dacarbazine and its photoproducts were separated on a Gemini C 18 reverse phase column 5 µm, 2.1 x 50 mm (Phenomenex, UK) fitted to a Perkin Elmer Series 200 Pump and UV/Vis Detector, and a 600 Series Link interface; all of which were controlled remotely using TotalChrom software (PerkinElmer, USA). Separation was achieved at a flow rate of 0.5 ml/min with a mobile phase of 70% water (adjusted to pH 3 with glacial acetic acid) and 30%
acetonitrile. An injection loop of 20 µl was used and the detector wavelength was set at 236 nm. The overall run time of the assay was 10 min. The retention times for DBZ and its photoproduct were 3.87 and 4.21 min, respectively. The linearity range of DBZ calibration graph ranged between 4.8 x 10 -5 M and 5.5 x 10 -4 M.

Maafi M, & Lee L.Y. (2015). Determination of Dacarbazine Φ-Order Photokinetics, Quantum Yields, and Potential for Actinometry. Journal of pharmaceutical sciences, 104(10).

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