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568223-U

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Discovery® BIO Wide Pore C18 (5 µm) HPLC Columns

L × I.D. 25 cm × 4.6 mm

Synonym(s):

Wide Pore C18 HPLC Column

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About This Item

UNSPSC Code:
41115700
eCl@ss:
32110501
NACRES:
SB.52
Pricing and availability is not currently available.

Product Name

Discovery® BIO Wide Pore C18 HPLC Column, 5 μm particle size, L × I.D. 25 cm × 4.6 mm

material

stainless steel column

Quality Level

Agency

suitable for USP L1

product line

Discovery®

feature

endcapped

manufacturer/tradename

Discovery®

packaging

1 ea of

extent of labeling

9.2% Carbon loading

parameter

0-70 °C temperature
400 bar pressure (5801 psi)

technique(s)

HPLC: suitable
LC/MS: suitable

L × I.D.

25 cm × 4.6 mm

surface area

100 m2/g

surface coverage

3.6 μmol/m2

matrix

silica particle platform
fully porous particle

matrix active group

C18 (octadecyl) phase

particle size

5 μm

pore size

300 Å

operating pH

2-8

separation technique

reversed phase

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General description

Peptide maps generated by RP-HPLC provide valuable information about protein structure, stability, and purity. To be effective, the RP-HPLC column must be able to resolve a high percentage of the peptides in the sample. The more peptides, the better the information. Discovery BIO Wide Pore C18 gives unsurpassed RP-HPLC resolution of peptide maps from enzymatic digests. The improvements in silica and bonded-phase chemistry we have incorporated into the Discovery BIO Wide Pore line improve resolution by increasing efficiency and reducing the peak tailing. An added benefit to this is the ability to analyze peptides without TFA in the mobile phase, thereby increasing the LC-MS signal.

Application


  • Kinetic performance comparison of superficially porous, fully porous and monolithic reversed-phase columns by gradient kinetic plots for the separation of protein biopharmaceuticals.: This study provides a detailed kinetic performance comparison of different types of reversed-phase HPLC columns, including the Discovery® BIO Wide Pore C18 HPLC Column. The research highlights its efficiency and resolution in separating complex protein biopharmaceuticals, making it a valuable tool for analytical chemists in biopharmaceutical analysis (Jaag et al., 2022).

  • RP-LC x RP-LC analysis of a tryptic digest using a combination of totally porous and partially porous stationary phases.: This paper investigates the use of reverse-phase liquid chromatography (RP-LC) with both totally porous and partially porous stationary phases, including the BIO Wide Pore C18 HPLC Column, to analyze tryptic digests. The findings demonstrate its robustness and high resolution in proteomics applications, enabling precise peptide mapping and protein characterization (Mondello et al., 2010).

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Legal Information

Discovery is a registered trademark of Merck KGaA, Darmstadt, Germany

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Hannah F Speer et al.
Journal of animal science, 98(9) (2020-08-28)
This study assessed the relative bioavailability of guanidinoacetic acid (GAA) in cattle. Seven ruminally cannulated Holstein steers (initial body weight of 280 kg) were used in an experiment with a 5 × 5 Latin square design; the two additional steers

Related Content

DiscoveryBIO Wide Pore C18 products offered by Sigma-Aldrich.

Chromatograms

application for HPLC

Questions

1–2 of 2 Questions  
  1. What is the method for rewetting a column that may have dried as a result of the end cap being loose or not being used for an extended period of time?

    1 answer
    1. Reverse Phase Column Rehydration Method:
      1. The column should ideally not contain any buffer. If it does, thoroughly flush the column with water to remove the buffer.
      2. Flush the column with 100% acetonitrile, starting at a flow rate of 0.3-0.5 ml/minute. Slowly increase the flow rate to 1 ml/minute, flushing for approximately 30 minutes, as long as the pressure does not excessively increase.
      3. Equilibrate the column with the mobile phase specified on the Certificate of Analysis (CofA) of the column, and run a test mix similar to what was used to test the column.
      4. If the performance is unsatisfactory, repeat the method, increasing the flushing with acetonitrile for one hour.
      5. Repeat the test. If the performance is still not as expected, the column may have degraded.

      Helpful?

  2. WHIGH SOLVENT USED FOR WASHING OF COLUMN

    1 answer
    1. See below for a general cleaning protocol:
      1. Reverse the orientation of the column
      2. Flush for a few minutes, or >1 column volume, with a 20% solution in water, of the organic used in the analysis, such as methanol or acetonitrile. This is to remove any residual buffer salts.
      3. Flush for approximately 30 mins, or >10 column volumes, with 100% of the organic solvent used in the analysis.
      4. The column can be stored with this organic solvent until ready for use.
      5. Before reusing the column, flush again, as per step 2.

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