Evoc18 column

Manufactured by Phenomenex

Sourced in France

The EvoC18 column is a reversed-phase high-performance liquid chromatography (HPLC) column designed for the separation and analysis of a wide range of organic compounds. It features a C18 stationary phase with optimized pore size and surface area for efficient chromatographic performance.

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Lab products found in correlation

Ultimate 3000 uplc, by Thermo Fisher Scientific (4 mentions) Lc 20ar pumps, by Phenomenex (2 mentions) Spd 20a uv vis detector, by Phenomenex (2 mentions) Whatman 1ps phase separator, by GE Healthcare (2 mentions) Whatman, by Cytiva (2 mentions) Uhplc system, by Shimadzu (1 mentions) Hplc system, by Shimadzu (1 mentions)

Close spelling variants of this product

Kinetex 1.7 μm EVOC18 100 Å LC column (2 citations)

9 protocols using evoc18 column

HPLC Purification of Peptides and Amphiphiles

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HPLC purification
was performed on a Shimadzu system equipped with two LC-20AR pumps,
an SPD-20A UV–vis detector, and a Phenomenex Kinetex EVO C18
column. The mobile phases were water and acetonitrile, containing
either 0.1% TFA, for peptides, or 0.1% NH₃, for amphiphiles.
The purity of the compounds was assessed using LC-MS (Figures S15–S25). All purified molecules
were lyophilized and stored at −20 °C until required.

Egorova E.A., van Rijt M.M., Sommerdijk N., Gooris G.S., Bouwstra J.A., Boyle A.L, & Kros A. (2020). One Peptide for Them All: Gold Nanoparticles of Different Sizes Are Stabilized by a Common Peptide Amphiphile. ACS Nano, 14(5), 5874-5886.

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Aflatoxin B1 Quantification by HPLC

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AFB1 analysis was done as described by El Khoury et al. [19 (link)]. In brief, AFB1 extraction was done with 30 mL of HPLC-grade absolute chloroform. Next, supernatants were filtered through a Whatman 1PS phase separator (GE Healthcare Life Sciences, Vélizy-Villacoublay, France). Two milliliters of filtrate were evaporated to dryness in a STUART SBH200D/3 sample concentrator (Stuart equipment, Paris, France) at 45 °C, and the samples were re-solubilized in 2 mL of acetonitrile. Finally, the samples were filtered through 0.45 μm disk filters (Thermo Fisher Scientific, Illkirch-Graffenstaden, France) and placed in HPLC vials. AFB1 was analyzed by using an Ultimate 3000 UPLC (Thermo Fisher Scientific, Illkirch-Graffenstaden, France) with an EvoC18 column (3 µm, 150 × 3.2, Phenomenex, Le Pecq, France) conditioned at 27 °C. The elution program used for separation consisted of forming an isocratic mixture composed of acetonitrile and water (25:75 v:v). The mobile phase had a flow rate of 1.2 mL/min. Ten microliters of sample were injected. AFB1 was detected by using a fluorescent detector at 365 (430) nm excitation (emission) wavelengths. The identity of the molecule was confirmed by analyzing the UV absorption spectrum by an additional diode matrix detector (DAD) coupled to the system. AFB1 production levels were calculated based on a standard calibration curve (0.16 to 20 mg/L).

Hernandez C., Cadenillas L., Maghubi A.E., Caceres I., Durrieu V., Mathieu C, & Bailly J.D. (2021). Mimosa tenuiflora Aqueous Extract: Role of Condensed Tannins in Anti-Aflatoxin B1 Activity in Aspergillus flavus. Toxins, 13(6), 391.

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Bioactive Molecule Characterization in Lemon Verbena

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The characterization of the bioactive molecules was done as described by Quirantes- Piné et al. [37 (link)]. Analyses were carried out on an Ultimate 3000 UPLC (Thermo-Fisher, Illkirch-Graffenstaden, France), including a diode array detector (DAD). An EvoC18 column (2.6 µm, 150 × 2.1 mm, Phenomenex, Le Pecq, France) was used for all assays. The mobile phases consisted of solvents A and B, which were respectively water: acetonitrile (90:10 v/v) with 1% of formic acid and acetonitrile. The linear gradient applied was 5% B 0–25 min, 20% B 25–30 min, 40% B 30–35 min, 5% B 35–40 min. The separation was performed at 30 °C with a flow rate of 0.5 mL/min. The injection volume of each sample was 20 µL. The UV detection was made in the λ range of 190–450 nm.
To better identify some of the components of the A. citrodora extract, the following standards were used: luteolin-7-diglucuronide, verbascoside, apigenin-7-glucuronide and caffeic acid. Standards were dissolved in Dimethyl sulfoxide (DMSO) to obtain a final concentration of 700 mg/L. Different concentrations of the standards were injected (50–100–200–300–400–500 mg/L) in order to draw calibration curves.

Cadenillas L.F., Hernandez C., Bailly S., Billerach G., Durrieu V, & Bailly J.D. (2023). Role of Polyphenols from the Aqueous Extract of Aloysia citrodora in the Inhibition of Aflatoxin B1 Synthesis in Aspergillus flavus. Molecules, 28(13), 5123.

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Quantifying Brain S1P Levels

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S1P levels were measured as previously described [32 (link)] with some modifications. Briefly, 20 μl of brain homogenate was suspended in 200 μl methanol with 0.25% conc. HCl. Alkaline extraction of the lipids was performed by addition of 400 μl chloroform, 30 μl of 10 M NaOH, 580 μl of 2 M KCl and 200 μl methanol with 200 pmol C17-S1P and centrifuged (5 min, 16000xg). The upper aqueous/methanol phase was collected and acid extraction was performed with 40 μl conc. HCl and 300 μl chloroform. The upper phase was aspirated and 200 μl of lower choloroform phase was collected and chloroform was evaporated using a speed vacuum system. The dried lipids were resuspended in 275 μl methanol/70 mM K₂HPO₄ (9:1) with 1 mM EDTA by sonication in a bath sonicator for 30 sec. A derivatization mixture of 10 mg o-phthalaldehyde, 200 μl ethanol, 10 μl β-mercaptoethanol and 10 ml of a 3% boric acid solution (adjusted to pH 10.5 with KOH) was prepared and 25 μl of this was added to the lipid samples and incubated for 15 min at room temperature followed by centrifugation. S1P levels were then determined by HPLC analysis with an EVO C18 column (Phenomenex, Lane Cove, NSW, Australia) as previously described [32 (link)].

Al-Shujairi W.H., Clarke J.N., Davies L.T., Alsharifi M., Pitson S.M, & Carr J.M. (2017). Intracranial Injection of Dengue Virus Induces Interferon Stimulated Genes and CD8+ T Cell Infiltration by Sphingosine Kinase 1 Independent Pathways. PLoS ONE, 12(1), e0169814.

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UHPLC-MS Analysis of CQAs and diCQAs

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For CQAs and diCQAs the samples were run on a Nexera/Prominence UHPLC system attached to a 2020 single quadrupole mass spectrometer (both from Shimadzu). Separation was on a 100×2.1mm 2.6μ EVO C18 column (Phenomenex) using the following gradient of acetonitrile (B) versus 0.1% formic acid in water (A), run at 0.55 ml.min^-1 and 40°C: 5% B over 0.0-11.0 min, 25% B over 11.0-11.5 min, 80% B over 11.5-14.0 min, 5% B over 14.0-18.00 min. Samples were kept at 15°C in the autosampler prior to injection. The PDA collected spectra from 200-800 nm at 6.25Hz with a time-constant of 0.24 sec. The MS was equipped with a dual ion source, used in electrospray mode (no APCI), with spray chamber conditions of 250°C desorbation line temperature, 200°C heat block, 1.5l.min^-1 nebulizer gas, and 15l.min^-1 drying gas.
CQAs and diCQAs were analysed by single ion monitoring event in negative mode monitoring the following ions for 0.25 sec in total: m/z 353 for CQA and isomers and m/z 515 for diCQAs.

D’Orso F., Hill L., Appelhagen I., Lawrenson T., Possenti M., Li J., Harwood W., Morelli G, & Martin C. (2023). Exploring the metabolic and physiological roles of HQT in S. lycopersicum by gene editing. Frontiers in Plant Science, 14, 1124959.

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HPLC-based Aflatoxin B1 Quantification

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AFB1 was extracted from culture media with HPLC-grade chloroform and the supernatants were filtered off. Filtrates (2 mL) were concentrated to dryness at 45 °C using a STUART SBH200D/3 sample concentrator (Stuart equipment, Paris, France), and the residues were redissolved in acetonitrile HPLC grade (2 mL). Finally, the samples were filtered through 0.45 µm PTFE disk filters (Thermo Fisher Scientific, Illkirch-Graffenstaden, France) and placed in HPLC vials. AFB1 concentration was quantified by HPLC on an Ultimate 3000 UPLC equipment coupled to fluorescent detector at 365 nm wavelengths for excitation (and 430 nm for emission) (Thermo Fisher Scientific, Illkirch-Graffenstaden, France). Separation was carried out with an EvoC18 column (3 µm, 150 × 3.2, Phenomenex, Le Pecq, France) at 27 °C, at a 1.2 mL/min flow of a mixture acetonitrile HPLC grade and water (25:75 v:v). Injection volume was 10 µL.
The identity of the AFB1 was confirmed by checking the UV absorption spectrum thanks to an additional DAD detector in the series. AFB1 concentrations were calculated with an external standard calibration curve in the range of 0.16 to 20 mg/L.

Hernandez C., Cadenillas L., Mathieu C., Bailly J.D, & Durrieu V. (2022). Preservation of Mimosa tenuiflora Antiaflatoxigenic Activity Using Microencapsulation by Spray-Drying. Molecules, 27(2), 496.

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HPLC Purification of Organic Compounds

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A Shimadzu HPLC system equipped with two LC-20AR pumps, an SPD-20A UV-Vis detector, and a Phenomenex Kinetex EVO C18 column (21.2 by 150 mm) was used for purification. The mobile phases were water and acetonitrile (MeCN), containing either 0.1% ammonia (for 1-3 and 1-W, 2-W, 3-W) or 0.1% TFA (for 4, 1-K, and 2-K). Prior to purification of 1-3, the protecting TNP group was removed by incubation with tris(2-carboxyethyl) phosphine hydrochloride (TCEP, added in excess, 30 min). The purity of the compounds was assessed using LC-MS (Fig. S1-S9, ESI †).
All purified molecules were lyophilised and stored at À20 1C until needed.

Egorova E.A., Arias-Alpizar G., Vlieg R.C., Gooris G.S., Bouwstra J.A., Noort J.V., Kros A, & Boyle A.L. (2022). Coating gold nanorods with self-assembling peptide amphiphiles promotes stability and facilitates in vivo two-photon imaging. Journal of materials chemistry. B, 10(10).

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Quantification of Aflatoxin B1 in Cultures

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AFB1 extraction and quantification were done as previously described [15 (link)]. Culture media were mixed with 30 mL of HPLC-grade absolute chloroform and shaken at 200 rpm for 1 h at room temperature. Supernatants were filtered through a Whatman 1PS phase separator (GE Healthcare Life Sciences, Vélizy-Villacoublay, France). Two mL of the filtrates were evaporated to dryness in a STUART SBH200D/3 sample concentrator (Stuart equipment, Paris, France) at 45 °C and redissolved in 2 mL of acetonitrile. Furthermore, they were filtered through 0.45 μm PTFE disk filters (Thermo Fisher Scientific, Illkirch-Graffenstaden, France) and placed in HPLC vials. AFB1 quantification was done using an Ultimate 3000 UPLC (Thermo-Fisher Scientific, Illkirch-Graffenstaden, France) with an EvoC18 column (3 µm, 150 × 3.2 mm, Phenomenex, Le Pecq, France) conditioned at 27 °C. In order to separate AFB1, an elution program consisting of an isocratic mixture of acetonitrile and water (25:75 v:v) was used. The mobile phase had a flow rate of 1.2 mL/min and 10 µL of the sample was injected. Aflatoxin B1 was detected by using a fluorescent detector at 365 (430) nm excitation (emission) wavelengths. A diode matrix detector (DAD) coupled to the system was used to analyze the UV spectra. AFB1 levels were calculated based on a standard calibration curve in a range of 0.16 to 20 µg/mL.

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Purification of Compound SoL A by HPLC

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The active subfraction from Sephadex LH-20 column was purified by semi-preparative HPLC using a Phenomenex Kinetex EVO-C18 Column (100 Å, 5μm, 10×250 mm) and eluted at 3.8 ml/min in a gradient of solvents A (0.1% NH₄OH in water) and B (methanol): 80% B for 20 min to afford SoL A (1.63 mg). The concentration of SoL A from the cultivation was estimated as ~9.0 mg/L (14.5 μM) by a comparative LC-MS analysis between the crude extract and isolated pure SoL A.

Hou L., Tian H.Y., Wang L., Ferris Z.E., Wang J., Cai M., Older E.A., Raja M.R., Xue D., Sun W., Nagarkatti P., Nagarkatti M., Chen H., Fan D., Tang X, & Li J. (2022). Identification and biosynthesis of pro-inflammatory sulfonolipids from an opportunistic pathogen Chryseobacterium gleum. ACS chemical biology, 17(5), 1197-1206.

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