Zorbax 300extend c18

Manufactured by Agilent Technologies

Sourced in Japan, United States

The ZORBAX 300Extend-C18 is a reversed-phase high-performance liquid chromatography (HPLC) column designed for the separation and analysis of a wide range of analytes. It features a 300 Å pore size and a C18 stationary phase, providing a durable and reliable platform for various applications.

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Lab products found in correlation

1260 hplc system, by Agilent Technologies (1 mentions) Tmt 10 plex reagent, by Thermo Fisher Scientific (1 mentions) Ultimate 3000, by Thermo Fisher Scientific (1 mentions) Isolute c18, by Biotage (1 mentions)

8 protocols using zorbax 300extend c18

HPLC analysis of vitamin D2 and ergosterol

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A volume of 10 μL of filtered sample was
injected into an HPLC system equipped with 1260 quat pump, 1290 vial
sample, 1260 TCC column oven, and 1260 DAD VL detector (Agilent Co.,
Santa Clara) and was eluted through a reversed-phase C18 column (Zorbax
300 extend-C18, 4.6 × 100 mm² (3.5 μm), Agilent
Co., Santa Clara). The mobile phase was 0.1% formic acid/acetonitrile,
5:95, at a flow rate of 1 mL/min, and UV detection was performed at
264 and 282 nm. Vitamin D₂ and ergosterol were determined
by comparing the retention times to those of the standards, and quantification
was conducted by using a calibration curve.
The two compounds
vitamin D₂ and ergosterol were well separated, with retention
times of 5.8 and 7.8 min, respectively. Quantification of vitamin
D₂ and ergosterol was achieved by extrapolation from a
standard curve. The eight-point calibration curves used for quantification,
which were obtained using a linear fit, ranged from 1 to 200 μg/mL
(y = 17.19x – 1.30) and from
10 to 100 μg/mL (y = 10.69x + 4.79) and had r² values of 0.9999
and 0.9998 for vitamin D₂ and ergosterol, respectively.
The average recoveries (n = 9) of vitamin D₂ and ergosterol were 98.69 and 100.31% with average relative standard
deviations (RSDs) (n = 9) of 2.91 and 2.69%, respectively.

Hu D., Chen W., Li X., Yue T., Zhang Z., Feng Z., Li C., Bu X., Li Q.X., Hu C.Y, & Li L. (2020). Ultraviolet Irradiation Increased the Concentration of Vitamin D2 and Decreased the Concentration of Ergosterol in Shiitake Mushroom (Lentinus edodes) and Oyster Mushroom (Pleurotus ostreatus) Powder in Ethanol Suspension. ACS Omega, 5(13), 7361-7368.

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HPLC Analysis of Compound TFH

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The HPLC analysis was performed on a Shimadzu LC-20AT system (Tokyo, Japan) equipped with an Agilent Zorbax 300Extend C18 (150×4.6 mm, 5 μm) column. The mobile phase was set 0.8 mL/min at 30°C. The mobile phase consisted of acetonitrile (A) and 0.1% formic acid (B) was performed as follows: 5% (A) to 35% (A) for 20 min, and 35% (A) held for 10 min, and TFH was measured at 280 nm.

Liu F., Zhang X, & Ji Y. (2020). Total Flavonoid Extract from Hawthorn (Crataegus pinnatifida) Improves Inflammatory Cytokines-Evoked Epithelial Barrier Deficit. Medical Science Monitor : International Medical Journal of Experimental and Clinical Research, 26, e920170-1-e920170-8.

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Peptide Fractionation via Reverse-Phase LC

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Basic reverse-phase LC for fractionation of peptide mixture was performed with a 4.6 × 250–mm analytical column containing 5 μm of C18 particles (ZORBAX 300Extend-C18, Agilent). The flow rate was set as 1 mL/min. The mobile-phases were as follows: phase A,5 mM ammonium bicarbonate in water (pH 8.0), and phase B, 25 mM ammonium bicarbonate in water (pH 8.0) with 80% ACN. The gradient was as follows: held at 2% B for 5 min, from 2% to 5% B in 1 min, from 5% to 10% B in 9 min, from 10% to 30% B in 60 min, from 30% to 40% B in 15 min, from 40% to 80% B in 5 min, and held at 80% B for 20 min. Carboxypeptidase B–treated peptides (1.1 mg) were fractionated. A total of 51 fractions from 19.5 min to 96 min were collected and further mixed into 17 fractions. These 17 fractions were dried and desalted. Arginine dimethylated peptides in each fraction were then enriched by SECEM.

Wang Q., Li Z., Zhang S., Li Y., Wang Y., Fang Z., Ma Y., Liu Z., Zhang W., Li D., Liu C, & Ye M. (2022). Global profiling of arginine dimethylation in regulating protein phase separation by a steric effect–based chemical-enrichment method. Proceedings of the National Academy of Sciences of the United States of America, 119(43), e2205255119.

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Formaldehyde and Aniline Detection Protocol

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The determination of formaldehyde
was accomplished by the high-performance liquid chromatography (HPLC)
system (1260, Agilent Co., Santa Clara). It was eluted through a reversed-phase
C18 column (Zorbax 300 extend-C18, 4.6 × 100 mm (3.5 μm),
Agilent Co., Santa Clara). The mobile phase was water/acetonitrile,
45:55, at a flow rate of 1 mL/min, and UV detection was performed
at 413 nm. The column temperature was at 40 °C, and the injection
volume was 20 μL. The GC/MS analysis of aniline was performed
on a Thermo Trace gas chromatograph (Thermo Finnigan, Miami, FL) coupled
with Ultral ITQ 900 mass spectrometer (Thermo Finnigan, Miami, FL)
under the following analytical conditions: DB-WAX column (15 m ×
0.53 mm × 0.25 μm film thickness); helium (0.5 mL/min);
programmed temperature 50–220 °C (7 °C/min); injector
temperature (200 °C); and interface (230 °C).^{8 (link)}

Hu D., Yang X., Chen W., Feng Z., Hu C., Yan F., Chen X., Qu D, & Chen Z. (2021). Rhodiola rosea Rhizome Extract-Mediated Green Synthesis of Silver Nanoparticles and Evaluation of Their Potential Antioxidant and Catalytic Reduction Activities. ACS Omega, 6(38), 24450-24461.

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RP-HPLC Fractionation and Antimicrobial Analysis

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Fractions were obtained from filtered permeates by RP-HPLC (1100 series; Agilent Technologies Japan Ltd.). Separation was carried out with a Zorbax 300 Extend-C18 (250 × 4.6 mm, 5-µm particle size, 300-A pore size) from Agilent Technologies Japan Ltd. with a solvent flow rate of 0.25 mL/min. Once the column was equilibrated with solvent A (0.04% trifluoroacetic acid in water), 20 µL of the sample was injected. Fractions were eluted with an increasing gradient of solvent B (0.03% trifluoroacetic acid in acetonitrile) from 0 to 100% in solvent A for 70 min. Fractions monitored at 214 nm were collected from 15 chromatographic runs, freeze-dried, and resuspended in 1.5 mL of distilled water (Rodríguez-Figueroa et al., 2012) (link) for protein determination by Lowry method. Samples were adjusted to 0.30 mg/mL protein for antimicrobial activity determination and MS analysis.

Heredia-Castro P.Y., Reyes-Díaz R., Rendón-Rosales M.Á., Beltrán-Barrientos L.M., Torres-Llanez M.J., Estrada-Montoya M.C., Hernández-Mendoza A., González-Córdova A.F, & Vallejo-Cordoba B. (2021). Novel bacteriocins produced by Lactobacillus fermentum strains with bacteriostatic effects in milk against selected indicator microorganisms. Journal of dairy science, 104(4).

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Peptide Fractionation using HPLC

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Peptides (100 µg) were fractionated using a ZORBAX 300 Extend-C18 2.1 × 150 mm, 3.5 µm column on a 1260 HPLC system (Agilent, Santa Clara, CA, USA). Buffer A (5 mM ammonium formate (NH₄COOH)) and B (5 mM NH₄COOH, 90% acetonitrile in water) were used for the fractionation at a constant flow rate of 0.3 ml/min.

Ahn S.B., Sharma S., Mohamedali A., Mahboob S., Redmond W.J., Pascovici D., Wu J.X., Zaw T., Adhikari S., Vaibhav V., Nice E.C, & Baker M.S. (2019). Potential early clinical stage colorectal cancer diagnosis using a proteomics blood test panel. Clinical Proteomics, 16, 34.

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Quantification of Vitamin D2 and Ergosterol

Cited 2 times

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Vitamin D₂ and ergosterol were extracted and analyzed using a modified method, as reported previously [6 (link),7 (link)]. The standard curve was obtained using series standard solutions of vitamin D₂ and ergosterol. The quantification of vitamin D₂ and ergosterol was achieved by extrapolation from a standard curve. The samples of mushroom powder (0.2 g) were ultrasonically extracted with 3 mL of ethanol for 25 min at 50 °C, three times. The ethanol phase was then collected and diluted with ethanol to 10 mL. The extract was filtered through a 0.45 μm filter, before undergoing high-performance liquid chromatography (HPLC) analysis. A volume of 10 μL of filtered sample was directly injected into an HPLC system (1260, Agilent Co., Santa Clara) and was eluted through a reversed-phase C18 column (Zorbax 300 extend-C18, 4.6 × 100 mm (3.5 μm), Agilent Co., Santa Clara, CA, USA). The mobile phase for HPLC was acetonitrile/0.1% formic acid (95:5, v/v) in a constant program at a flow rate of 1.0 mL/min. The UV detection was set at 264 nm.

Hu D., Wang Y., Kong F., Wang D., Hu C., Yang X., Chen X., Chen W, & Feng Z. (2024). Analysis of Volatile Aroma Components in Different Parts of Shiitake Mushroom (Lentinus edodes) Treated with Ultraviolet C Light-Emitting Diodes Based on Gas Chromatography–Ion Mobility Spectroscopy. Molecules, 29(8), 1872.

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TMT-based Quantitative Proteomics

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For TMT experiment, first 10 μg of each peptide sample was pooled for the reference peptide channel. Ten TMT 10plex reagents (Thermo Fisher) with different mass of reporter ions were labeled on N-terminus or amine group of Lysine amino acid, and the last channel of TMT-131 was used for reference peptides to normalize experimental batch effects. After quenching of excess TMT reagent using 5 % ammonium hydroxide (NH₄OH), all channels in a batch were pooled in a tube followed by cleaning up salts using C18 column (Isolute-C18, Biotage). Eluted peptides were fractionated on HPLC (Ultimate 3000, Thermo Fisher) coupled with C18 column (ZORBAX 300Extend-C18, Agilent). Mobile phases were composed of 2% ACN, 15 mM NH₄OH, pH 10 for Buffer A and 90% ACN, 15 mM NH₄OH, pH 10 for Buffer B. Sample separation was accomplished using the following linear gradient: from 0% to 7% B in 2 min, from 7% to 45% B in 78 min, from 45% to 80% B in 5 min, and held at 80% B for an additional 10 min. Separated samples were collected in a 96 deep well plate and further concatenated into 24 fractions. The fractionated peptides were dried down and reconstituted in 0.1% FA with 3% ACN at a final concentration of 0.1 μg/μL for further analysis.

Woo J., Sudhir P.R, & Zhang Q. (2020). Pancreatic tissue proteomics unveils key proteins, pathways, and networks associated with type 1 diabetes. Proteomics. Clinical applications, 14(6), e2000053.

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