C18 sphereclone

Manufactured by Phenomenex

Sourced in United States

The C18 SphereClone is a reversed-phase liquid chromatography column designed for the separation and analysis of a wide range of organic compounds. It features a spherical silica-based stationary phase with chemically bonded C18 alkyl chains, providing efficient and reproducible chromatographic performance.

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Lab products found in correlation

Uflc pda, by Shimadzu (2 mentions) Luna c8, by Phenomenex (1 mentions) Scl 10a system, by Shimadzu (1 mentions) Lc 2010a system, by Shimadzu (1 mentions) Whatman no 4 paper, by Cytiva (1 mentions) Ultra fast liquid chromatograph, by Shimadzu (1 mentions)

6 protocols using c18 sphereclone

RP-HPLC Purification of Peptides and Glycopeptides

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RP-HPLC analysis and semi-preparative purifications were performed as follows:
Analytical separations were done in an LC-2010A system (Shimadzu, Kyoto, Japan) using Luna C₈ (3 µm, 50 mm × 4.6 mm) and Sphereclone C₁₈ (5 µm, 250 mm × 10 mm) columns (Phenomenex, Torrance, CA, USA) for peptides and glycopeptides, respectively. Linear gradients of solvent B (0.036% TFA in ACN) into solvent A (0.045% TFA in H₂O) were used over 15 and 20 min, respectively, at a 1 mL/min flow rate.
Semi-preparative purifications were done in a SCL-10A system (Shimadzu) using Phenomenex Luna C₈ (10 µm, 250 mm × 10 mm) and SphereClone C₁₈ (10 µm, 250 mm × 10 mm) columns for peptides and glycopeptides, respectively. Linear gradients of solvent B (0.1% TFA in ACN) into A (0.1% TFA in H₂O) over 30 and 20 min, respectively, were used, at a 5 mL/min flow rate.

Defaus S., Avilés M., Andreu D, & Gutiérrez-Gallego R. (2018). Lectin-Binding Specificity of the Fertilization-Relevant Protein PDC-109 by Means of Surface Plasmon Resonance and Carbohydrate REcognition Domain EXcision-Mass Spectrometry. International Journal of Molecular Sciences, 19(4), 1076.

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Synthesis and Purification of Glycopeptides

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Conjugation between N[Me]-O-Aoa-GFKKG-amide peptide and oligosaccharides was done at 20 and 25 mM, respectively in 0.1 M NaOAc for 72 h at 37 °C and pH 3.5 for NAc-hexosamines or pH 4.6 for hexoses. Conjugations involving Fuc-containing disaccharides (Fuc-α1,(3,4,6)-GlcNAc) were performed both in the presence and the absence of 100 mM aniline as a nucleophile catalyst, to explore improvements in conjugation yields.
All glycoconjugates were purified by semi-preparative RP-HPLC on SphereClone C₁₈ (Phenomenex, 250 mm × 10 mm; 5 μm) using a 10–20% linear gradient of acetonitrile into water (both eluents with 0.1% TFA). Immediately after purification, glycopeptide-containing fractions were neutralized with 10 mM NH₄HCO₃ (up to pH~5) to prevent acid degradation, and lyophylized. All synthetic di/trisaccharide-N[Me]-O-Aoa-GFKKG-amide glycopeptides were >90% pure by analytical RP-HPLC and had the expected mass by MALDI-TOF MS as shown Table 5.

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Quantitative Analysis of Organic Acids

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The assessment of organic acids, as earlier defined by the authors [42 (link)], was made via ultra-fast liquid chromatography combined with a photodiode array detector (UFLC-PDA; Shimadzu Corporation, Kyoto, Japan), and the chromatographic separation occurred in a C18 SphereClone (Phenomenex, Torrance, CA, USA) reverse-phase column (5 µm, 250 × 4.6 mm i.d.) thermostated at 35 °C, utilizing 3.6 mM sulfuric acid solution as an eluent at a flow rate of 0.8 mL/min. The classification was performed by making a comparison of the chromatograms obtained for the analyzed samples with commercial standards. The quantification of organic acids was carried out by relating the peak areas recorded at 215 nm, with the calibration curves obtained with authentic commercial standards for each compound. The outcomes were stated in g/100 g dw.

Baptista E., Liberal Â., Cardoso R.V., Fernandes Â., Dias M.I., Pires T.C., Calhelha R.C., García P.A., Ferreira I.C, & Barreira J.C. (2024). Chemical and Bioactive Properties of Red Rice with Potential Pharmaceutical Use. Molecules, 29(10), 2265.

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Determination of Organic Acids in Samples

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Organic acids were determined following a previously described procedure and optimized by the authors [1 (link)]. In brief, samples (~1.5 g) were extracted by stirring with 25 mL of metaphosphoric acid (25 °C at 60 g) for 25 min, and subsequently filtered through Whatman no. 4 paper (diam. 12.5 cm). The assessment was achieved by ultra-fast liquid chromatography coupled with a photodiode array detector (UFLC-PDA; Shimadzu Corporation, Kyoto, Japan). The compound separation was carried out in a C18 SphereClone (Phenomenex, Alcobendas, Spain) reverse phase column (5 µm, 250 × 4.6 mm id) thermostated at 35 °C using 3.6 mM sulfuric acid (0.02%) solution as an eluent at a flow rate of 0.8 mL/min. The identification was carried out by comparing the chromatograms obtained for the analyzed samples with those obtained using commercial standards. The quantification of the compounds was completed by relating the peak areas, recorded at 215 nm, with the calibration curves obtained with commercial standards for each compound. The results were expressed in g per 100 g of fw.

Liberal Â., Fernandes Â., Dias M.I., Pinela J., Vivar-Quintana A.M., Ferreira I.C, & Barros L. (2021). Phytochemical and Antioxidant Profile of Pardina Lentil Cultivars from Different Regions of Spain. Foods, 10(7), 1629.

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Quantitative Analysis of Organic Acids

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The analysis of organic acids followed the protocol established by the group [70 (link)]. The evaluation was performed by ultra-fast liquid chromatography coupled to a photodiode array detector (UFLC-PDA; Shimadzu Coperation, Kyoto, Japan). The separation of the compounds was carried out in a C18 SphereClone (Phenomenex) reverse phase column (5 μm, 250 × 4.6 mm id) thermostated at 35 °C, using 3.6 mM sulfuric acid solution as an eluent at a flow rate of 0.8 mL/min. The identification was carried out by comparing the chromatograms obtained for the analyzed samples with those obtained using commercial standards. The quantification of the compounds was done by relating the peak areas, recorded at 215 nm, with the calibration curves obtained with commercial standards for each compound. The results are presented in mg per 100 g dw.

Novais C., Pereira C., Molina A.K., Liberal Â., Dias M.I., Añibarro-Ortega M., Alves M.J., Calhelha R.C., Ferreira I.C, & Barros L. (2021). Bioactive and Nutritional Potential of Medicinal and Aromatic Plant (MAP) Seasoning Mixtures. Molecules, 26(6), 1587.

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Organic Acid Quantification via HPLC-DAD

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Organic acids were determined following a procedure previously described by Carocho, Barros, et al. (2015) . The detection and identification of these compounds relied on a Shimadzu (Shimadzu Corporation, Kyoto, Japan) ultra-fast liquid chromatograph, coupled to a diode array detector (DAD) with the wavelengths set at 215 and 245 nm (for ascorbic acid). The used column was a reverse phase C18 SphereClone (Phenomenex, Torrance, CA, USA). The compounds were identified by comparison with calibration curves of commercial standards. The equations were: (y = 9 Â 106x + 377,946; R2 = 0.994); quinic acid (y = 612,327x + 16 563; R2 = 1); malic acid (y = 863,548x + 55,591; R2 = 0.999); shikimic acid (y = 8 Â 107x + 55,079; R2 = 0.999); citric acid (y = 1 Â 106x + 16,276; R2 = 1); succinic acid (y = 603,298x + 4994.1; R2 = 1); fumaric acid (y = 148,083x + 96,092; R2 = 1). The results were expressed in mg per g of lyophilized decoctions.

Carocho M., Barros L., Barreira J.C., Calhelha R.C., Soković M., Fernández-Ruiz V., Buelga C.S., Morales P, & Ferreira I.C. (2016). Basil as functional and preserving ingredient in "Serra da Estrela" cheese. Food chemistry, 207.

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