Nucleodur c18 htec column

Manufactured by Macherey-Nagel

Sourced in Germany, Japan

Nucleodur C18 HTec 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 silica-based stationary phase with C18 bonded ligands, providing high resolution and efficient separations.

Automatically generated - may contain errors

Lab products found in correlation

Chromaster, by Hitachi (3 mentions) Dionex ultimate 3000, by Thermo Fisher Scientific (2 mentions) Reversed phase hplc, by Jasco (1 mentions) Ascend 700 mhz nmr spectrometer, by Bruker (1 mentions) 5 mm txi cryoprobe, by Bruker (1 mentions) Sephadex lh 20 column, by Merck Group (1 mentions) L2455 dad detector, by Hitachi (1 mentions) L 7420, by Hitachi (1 mentions) 2 ethylbutyric acid, by Merck Group (1 mentions) Polygoprep c18, by Macherey-Nagel (1 mentions) Plc 2020, by Gilson (1 mentions) Nexera uhplc system, by Shimadzu (1 mentions)

Close spelling variants of this product

EC250/4 Nucleodur C18 HTec column (2 citations) VP250/21 NUCLEODUR C18 HTec column (2 citations) C18 HTec column (3 citations) Nucleodur C18 column (19 citations) C18 column (10 citations)

11 protocols using nucleodur c18 htec column

Isolation and Purification of Salinichelins

Check if the same lab product or an alternative is used in the 5 most similar protocols

The analysis and purification of the crude extract was carried out on a Dionex Ultimate 3000 (Thermo Fisher Scientific, Waltham, MA, USA). For analysis, the instrument was equipped with a Nucleodur C18 HTec column (5 μm, 5.6 × 250 mm, Macherey Nagel, Dueren, Germany) and run in isocratic mode with 97% A (H₂O with 0.05% trifluoroacetic acid) and 3% B (MeCN with 0.05% trifluoroacetic acid) at a flow of 1 ml min^–1 for 26 min with ultraviolet detection at 230 nm.
For purification, a semi-preparative Nucleodur C18 HTec column (5 μm, 10 × 250 mm, Macherey Nagel) was used. The same method was applied with a flow of 6.25 ml min^–1. Fractions were collected with a fraction collector. Overall six fractions were collected corresponding to salinichelins A-F with retention times of 4–21 min. Based on amount and purity, salinichelins A to C were subjected to nuclear magnetic resonance spectroscopy (NMR) analysis.

Bruns H., Crüsemann M., Letzel A.C., Alanjary M., McInerney J.O., Jensen P.R., Schulz S., Moore B.S, & Ziemert N. (2017). Function-related replacement of bacterial siderophore pathways. The ISME Journal, 12(2), 320-329.

+ Open protocol

+ Expand

Enzymatic Conversion of Z-PAOx to PAN

Check if the same lab product or an alternative is used in the 5 most similar protocols

The activity was determined in modified protocol according to literature known methods^{32 (link)}. The assay solution contained 437.5 µL potassium phosphate buffer (50 mM, pH 7.0), 12.5 µL of Z-PAOx (200 mM solution in dimethyl sulfoxide, final concentration 5.0 mM), 50.0 µL of purified OxdB(C)6His or its variants (0.15 mg mL⁻¹; 3.75 µM), in a total volume of 500 µL. The reaction was carried out for 1 min at 30 °C with shaking at 900 rpm. Simultaneous addition of 400 µL of acetonitrile (ACN) and 100 µL of hydrochloric acid (0.1 M) stopped the reaction. After centrifugation (21,500×g, 10 min, 4 °C), the supernatant was transferred into HPLC vials and the conversion to PAN was measured by reversed phase-HPLC (Jasco, Nova Scotia, Canada) in comparison to a calibration curve. Measurements were conducted on a Nucleodur C18 HTec column (Macherey–Nagel, Düren, Germany) at 40 °C isocratic with water/ACN (70:30 v/v) as mobile phase and UV detection at 210 nm. Assays were conducted in triplicates for each variant.

Oike K., Sproß J., Matsui D., Asano Y, & Gröger H. (2021). Protein engineering of the aldoxime dehydratase from Bacillus sp. OxB-1 based on a rational sequence alignment approach. Scientific Reports, 11, 14316.

+ Open protocol

+ Expand

Characterization of Metabolites from Streptomyces albus

Check if the same lab product or an alternative is used in the 5 most similar protocols

S. albus ATGSal2P2::Tn14 was grown at 30 °C for 3 days in 6 × 500-mL flasks containing 50 mL of TSB, and pre-culture was used to inoculate 100 × 500-mL flasks containing 50 mL of NL19 media. Cultures were incubated at 30 °C for 5 days. Metabolites were extracted as described above. The extracts from biomass and the supernatant were combined and fractionated by size-exclusion chromatography on an LH 20 Sephadex column (Sigma-Aldrich, USA) using methanol as the solvent. The fractions were collected every 15 minutes., evaporated and dissolved in 0.5 mL of MeOH. Samples were further separated by preparative HPLC (Dionex UltiMate 3000, Thermo Fisher Scientific, USA) using a NUCLEODUR® C18 HTec column (250 × 10 mm, 5 µm) (Macherey-Nagel, Germany) with a linear gradient of solvent B (acetonitrile with 0.1% of formic acid) against solvent A (water with 0.1% of formic acid) at a flow rate of 4.5 mL/min at 45 °C. Compounds were separated using a gradient starting from 30% and increasing to 70% of B over 30 min. UV spectra were recorded with a DAD detector at 280 nm. Individual peaks were collected and analyzed by LC-MS as described above.
NMR spectra were acquired on a Bruker Ascend 700 MHz NMR spectrometer equipped with a 5 mm TXI cryoprobe (Bruker, USA). Deuterated CDCL₃ was used as a solvent and HSQC, HMBC and ¹H-¹H COSY spectra were recorded using standard pulse programs (Table 5S).

Ahmed Y., Rebets Y., Tokovenko B., Brötz E, & Luzhetskyy A. (2017). Identification of butenolide regulatory system controlling secondary metabolism in Streptomyces albus J1074. Scientific Reports, 7, 9784.

+ Open protocol

+ Expand

Measuring PET Degradation by DuraPETase

Check if the same lab product or an alternative is used in the 5 most similar protocols

The PET degradation rates for the original DuraPETase and its variants were tested using amorphous PET (2.4% crystallinity, determined by differential scanning calorimetry) with a thickness of 0.25 mm purchased from Goodfellow Cambridge Limited (Huntingdon, England). Substrate cut-outs with a diameter of 6 mm were incubated in 300 µl of 50 mM bicine NaOH buffer pH 9 containing 100 nM of the respective enzyme in microvolume reaction tubes. All reactions were performed as triplicates and were incubated at 50 °C or 52 °C and 300 rpm for 3 days. The reactions were stopped by removing the substrate. Three volumes of acetonitrile were added, and a centrifugation step (20,000 × g, 10 min) was performed. For each sample, 10 µl of the supernatant was injected into a LaChrom Elite HPLC system equipped with a L-2455 DAD detector (Hitachi, Chiyoda, Japan) and a Nucleodur C18 HTec column (Macherey–Nagel, Düren, Germany). The separation was performed isocratically at 40 °C with a flow rate of 0.5 ml/min for 8 min using 70% ddH₂O, 20% acetonitrile, and 10% formic acid. The signal was detected at 254 nm, and standards of the reaction products terephthalic acid (TCI, Tokyo, Japan) and 2-hydroxyethyl terephthalic acid (Activate Scientific, Prien am Chiemsee, Germany) were used to quantify the degradation rate.

Schreiber S., Gercke D., Lenz F, & Jose J. (2024). Application of an alchemical free energy method for the prediction of thermostable DuraPETase variants. Applied Microbiology and Biotechnology, 108(1), 305.

+ Open protocol

+ Expand

Quantification of Fecal Short-Chain Fatty Acids in Mice

Check if the same lab product or an alternative is used in the 5 most similar protocols

The levels of fecal SCFAs in mice were analyzed as described by Torii et al. [21 (link)], with some modifications. Briefly, after feeding the mice with appropriate diets for 10 weeks, their fecal samples were collected. SCFAs in feces were extracted using 70% ethanol solution. The extracted solution was mixed with 2-ethylbutyric acid (109959, Sigma-Aldrich Co., St. Louis, MO, USA) as an internal standard. Next, a reaction-assistive agent was added, and the mixture was reacted at 60 °C for 20 min. The reaction was terminated with potassium hydroxide (30,603, Sigma-Aldrich) solution; the reaction mixture was incubated at 60 °C for 20 min and then extracted with a phosphoric acid solution (B0992, Katayama Chemical Industries Co., Ltd., Osaka, Japan) and ether. The upper ether layer was collected and mixed with water for further extraction. Finally, an ether layer was obtained and air-dried. Methanol was added to dissolve fatty acid hydrazide present in the sample. The final product was analyzed using high-performance liquid chromatography (HPLC). HPLC was performed using a LaChrom L-7100 HPLC system (Hitachi Ltd., Tokyo, Japan) equipped with a NUCLEODUR C18 HTec column (MACHEREY-NAGEL, Düren, Germany) and a UV-VIS detector (L-7420, Hitachi).

Chen Y.T., Hsu A.H., Chiou S.Y., Lin Y.C, & Lin J.S. (2021). AB-Kefir Reduced Body Weight and Ameliorated Inflammation in Adipose Tissue of Obese Mice Fed a High-Fat Diet, but Not a High-Sucrose Diet. Nutrients, 13(7), 2182.

+ Open protocol

+ Expand

Peptide Fractionation for ACE Inhibition

Check if the same lab product or an alternative is used in the 5 most similar protocols

Caulerpa lentillifera protein hydrolysate was sequentially fractionated using successive chromatographic techniques to obtain fractions enriched in peptides with ACEI activity. The peptides in the CLP hydrolysate were separated on the basis of hydrophobicity, charge, size and polarity. About 20 µL of the hydrolysate dissolved in 5% ACN and 0.1% TFA in deionized water was fractionated by HPLC (Hitachi Chromaster, Tokyo, Japan) with a NUCLEODUR^® C₁₈ HTec column (4.6 mm × 250 mm; particle size of 5 µm, Macherey-Nagel, Düren, Germany). The mobile phase was arranged using mobile phase A (5% ACN containing 0.1% TFA) and B (95% ACN containing 0.1% TFA). The gradient was programmed in the following order: 0–50 min gradient elution from 0% B to 25% B; 50–53 min elution going from 25% B to 80% B; 53–58 min isocratic elution with 80% B; 58–60 min gradient from 80% B to 0% B at constant flow rate of 1 mL/min. The peptide mixture was monitored using UV absorptions at 214 nm. The hydrolysate was separated into 12 fractions by every 5 min. These fractions were collected, freeze dried, and kept at −20 °C for the following ACE inhibitory assay.

Joel C.H., Sutopo C.C., Prajitno A., Su J.H, & Hsu J.L. (2018). Screening of Angiotensin-I Converting Enzyme Inhibitory Peptides Derived from Caulerpa lentillifera. Molecules, 23(11), 3005.

+ Open protocol

+ Expand

Isolation and Purification of Herpetopanone

Check if the same lab product or an alternative is used in the 5 most similar protocols

The fermentation and extraction conditions were the same as described in the feeding experiments, except that H. aurantiacus 114-95^T was grown on a 20 L scale in VNY medium and that the cultures were only supplemented with non-labeled D-glucose. The extract that was obtained after the XAD-2 elution was dissolved in 20% (v/v) aqueous MeOH and fractionated by flash column chromatography over Polygoprep C18 (Macherey-Nagel) using an increasing concentration of MeOH in water. Fractions containing 1 were identified by LC–MS analysis, pooled and purified by reversed-phase HPLC on a Shimadzu UFLC liquid chromatography system equipped with a Nucleodur C18 HTec column (VP 250 × 10 mm, 5 μm; Macherey-Nagel) using a gradient from 10% (v/v) MeOH in H₂O (+ 0.1% trifluoroacetic acid) to 100% (v/v) MeOH over 30 min. The purity of the isolated herpetopanone determined by HPLC was >95% at all wavelengths (compared to sum of total peak areas).

Pan X., Domin N., Schieferdecker S., Kage H., Roth M, & Nett M. (2017). Herpetopanone, a diterpene from Herpetosiphon aurantiacus discovered by isotope labeling. Beilstein Journal of Organic Chemistry, 13, 2458-2465.

+ Open protocol

+ Expand

HPLC Analysis of Phenolic Compounds

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

Ethanolic extracts (20 μL), previously filtered using a 0.45 μm nylon membrane filter, were injected in the HPLC system (Hitachi Chromaster, Tokyo, Japan). The chromatographic separation was carried out with a NUCLEODUR^® C₁₈ HTec column (250 × 4.6 mm, the particle size of 5 μm, Macherey-Nagel, Düren, Germany). The mobile phase consisted of deionized water containing 0.1% trifluoroacetic acid (A) and methanol (B). The gradient was programmed in the following order: 90% A at 0–3 min, 70% A at 20 min, 60% A at 30 min, 40% A at 50 min, and 80% A at 60 min. The flow rate was 1.0 mL/min, and the column temperature was maintained at 25 °C. Phenolic compounds were detected by comparing their chromatographic behavior and monitoring UV absorption at 320 nm with authentic standards and reported data [5 (link),10 (link),19 (link)].

Le T.N., Luong H.Q., Li H.P., Chiu C.H, & Hsieh P.C. (2019). Broccoli (Brassica oleracea L. var. italica) Sprouts as the Potential Food Source for Bioactive Properties: A Comprehensive Study on In Vitro Disease Models. Foods, 8(11), 532.

+ Open protocol

+ Expand

Peptide Synthesis and Purification

Check if the same lab product or an alternative is used in the 5 most similar protocols

Peptides were synthesised in a 10 μmol scale (0.25 mmol/g) following the standard solid phase peptide synthesis (SPPS) methodology, using Fmoc-amino acids and Oxyma/DIC as coupling agents. Final deprotection and cleavage from the solid support was performed with 1.5 ml of cleavage cocktail: 94 TFA/1 TIS/ 2.5 DODT/2.5 H₂O for 3 h. Obtained peptides were purified at 25°C by preparative reverse phase (RP)-HPLC performed on a PLC 2020 personal purification system (Gilson) with a preparative Nucleodur C18 HTec-column (5 μm, 250 × 16 mm; Macherey Nagel) and a flow rate of 10 ml/min. Detection of the signals was achieved with a UV detector at 220 nm wavelength. The eluents were MilliQ H₂O and MeCN with addition of 0.1% TFA applied at a gradient of 5-40% MeCN.
Peptides were diluted and concentrations were determined according to [49 (link)]. The following concentrations were used in interaction assays: 3.5 μM, 7.0 μM, 14.0 μM (FOG1 peptides) and 17.5 μM, 35 μM (Ush peptides) in GST pulldown assays; 1.0 μM, 2.0 μM, 3.0 μM FOG1 peptides in immunoprecipitation assays.

Lenz J., Liefke R., Funk J., Shoup S., Nist A., Stiewe T., Schulz R., Tokusumi Y., Albert L., Raifer H., Förstemann K., Vázquez O., Tokusumi T., Fossett N, & Brehm A. (2021). Ush regulates hemocyte-specific gene expression, fatty acid metabolism and cell cycle progression and cooperates with dNuRD to orchestrate hematopoiesis. PLoS Genetics, 17(2), e1009318.

+ Open protocol

+ Expand

Phenolic Profiling of Ethanolic Extracts

Check if the same lab product or an alternative is used in the 5 most similar protocols

The phenolic composition of ethanolic extracts was analyzed using an HPLC system (Hitachi Chromaster, Tokyo, Japan) according to the method previously described [28 ]. Sample compounds were separated in a NUCLEODUR^® C₁₈ HTec column (250 × 4.6 mm, the particle size of 5 µm, Macherey-Nagel, Düren, Germany) at 25 °C by a flow rate of 1 mL/min. The mobile phase consisted of a mixture of 0.1% trifluoroacetic acid solution (A) and 100% methanol (B) with the following gradient: from 0–3 min, 10% B; from 3–20 min, 30% B; from 20–30 min, 40% B; from 30–50 min, 60% B; and from 50–60 min, 20% B. The eluted peaks were detected at 280, 320, and 360 nm. Phenolic compounds were identified and quantified by comparing the chromatographic behavior and retention times in specific UV spectra with external standards and reported data.

Le T.N., Sakulsataporn N., Chiu C.H, & Hsieh P.C. (2020). Polyphenolic Profile and Varied Bioactivities of Processed Taiwanese Grown Broccoli: A Comparative Study of Edible and Non-Edible Parts. Pharmaceuticals, 13(5), 82.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!