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.
568223-U
Discovery® BIO Wide Pore C18 (5 µm) HPLC Columns
L × I.D. 25 cm × 4.6 mm
Synonym(s):
Wide Pore C18 HPLC Column
About This Item
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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
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).
Legal Information
Application
guard cartridge
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Chromatograms
application for HPLC-
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-
Helpful?
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WHIGH SOLVENT USED FOR WASHING OF COLUMN
1 answer-
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.Helpful?
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