Long-term storage of silica-based columns is best in 100% acetonitrile. Columns may be safely stored for short periods (up to 3 or 4 days) in most common mobile phases. However, when using buffers, it is best to remove the salts to protect both the column and the HPLC equipment by first flushing the column with the same mobile phase without the buffer (e.g., when using 90/10 ACN/buffer, flush the column with 90/10 ACN/H2O) to eliminate any concern about salt precipitation or corrosion from the salts then flush the column with 100% acetonitrile for storage.Before storing the column, the end-fittings should be tightly sealed with the endplugs that came with the column to prevent the packing from drying.
53939-U
Ascentis® Express 90 Å HILIC (2.7 μm) HPLC Columns
L × I.D. 10 cm × 2.1 mm, HPLC Column
Synonym(s):
Core-shell (SPP) Fused Core Si HPLC column
About This Item
Recommended Products
Product Name
Ascentis® Express HILIC, 2.7 μm HPLC Column, 2.7 μm particle size, L × I.D. 10 cm × 2.1 mm
material
stainless steel column
Quality Level
Agency
suitable for USP L3
product line
Ascentis®
feature
endcapped: no
manufacturer/tradename
Ascentis®
packaging
1 ea of
parameter
≤100 °C temp. range
600 bar max. pressure (9000 psi)
technique(s)
HPLC: suitable
LC/MS: suitable
UHPLC-MS: suitable
UHPLC: suitable
L × I.D.
10 cm × 2.1 mm
surface area
135 m2/g
impurities
<5 ppm metals
matrix
Fused-Core particle platform
superficially porous particle
matrix active group
silica phase
particle size
2.7 μm
pore size
90 Å
operating pH
1-8
application(s)
food and beverages
separation technique
hydrophilic interaction (HILIC)
normal phase
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General description
Visit the Ascentis Express home page for more information on this new column technology.
Legal Information
Application
analytical column
guard cartridge
related product
required but not provided
Storage Class Code
11 - Combustible Solids
WGK
WGK 3
Flash Point(F)
Not applicable
Flash Point(C)
Not applicable
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Articles
A significantly improved HPLC-fluorescence method for DMB-NANA and -NGNA, and application of this method to compare 2 candidate biosimilar therapeutic proteins to their respective RMs.
A significantly improved HPLC-fluorescence method for DMB-NANA and -NGNA, and application of this method to compare 2 candidate biosimilar therapeutic proteins to their respective RMs.
A significantly improved HPLC-fluorescence method for DMB-NANA and -NGNA, and application of this method to compare 2 candidate biosimilar therapeutic proteins to their respective RMs.
A significantly improved HPLC-fluorescence method for DMB-NANA and -NGNA, and application of this method to compare 2 candidate biosimilar therapeutic proteins to their respective RMs.
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Chromatograms
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How should I store the Ascentis Express HILIC column?
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In HILIC separations, what happens if the sample is an aqueous matrix? Does it always have a negative effect?
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Yes, it would be highly preferential (especially in this case where you want partitioning to dominate) to inject in high organic. That said, you can 'get away' with it if the injection volume can be kept small - much like we can inject low volumes of stronger solvents in RP mode, if needed. What you will want to do to minimize impact is to get as much retention on the analytes of interest as you can, this helps give the sample solvent some time to dissipate and negate the effects.
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Can peptide or protein samples be analyzed using HILIC columns?
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Polar peptides are quite amenable to HILIC separations; however, our experience with larger peptides has been only minimally successful - mainly due to solubility issues. Proteins are even more difficult due to the same issue. An additional problem with proteins is that they are often multiply charged. When IEX is performed on multiply charged analytes, you often get what is referred to as a rolling effect where the analyte interacts with ionic sites on the surface in many different ways as it 'rolls' down the column; this produces broad and misshapen peaks.
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What flow rate should I use with Ascentis® Express HPLC Columns?
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Based on the minimum in the van Deemter curves, higher flows than 5um particle columns are required in order to maximize Ascentis Express column efficiency. The suggested starting point for flow rate for Ascentis Express columns: 1.6 mL/min for 4.6 mm ID; 0.8 mL/min for 3.0 mm ID; and 0.4mL/min for 2.1 mm ID.
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When you recommend only changing one parameter at a time, does this also refer to the total ionic strength?
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If you change organic, try to keep the overall buffer concentration (and all other parameters, for that matter) constant. There are times when you will want to change both and perhaps pH/temp/etc. simultaneously, but that drastically complicates the system and thus should be avoided, if possible.
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What is the Department of Transportation shipping information for this product?
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Transportation information can be found in Section 14 of the product's (M)SDS.To access the shipping information for this material, use the link on the product detail page for the product.
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Can I use Ascentis Express on any type of HPLC system?
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Ascentis Express HPLC columns are capable of use on standard HPLC systems as well as UHPLC systems. Columns are packed in high pressure hardware capable of withstanding the pressures used in UHPLC systems.
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Why is it recommended to run isocratically for HILIC methods?
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When running in HILIC mode, both isocratic and gradient practices result in instability. If you keep the re-equilibration times constant, gradient should not be a problem, but changing this parameter can have a significant impact. It is not so much that it is bad as it is different than we are used to in reversed phase. Usually, we assume that once equilibrated (5, 10, 15 min, etc.), we can leave the system for any time period and come back to the same results. This does not appear to be the case in our studies of HILIC. Knowing that the re-equilibration time has an impact, you should get in the habit of making several injections with known re-equilibration times prior to making any development decisions. To get around this, isocratic runs are recommended. Attached are two posters; the first was presented at HPLC 2013 (Amsterdam) and the second was presented at Balaton Symposium on High Performance Separation Methods 2013 (Hungary). Both show 'reproducibility' at any set re-equilibration time is good but both show that if you change the re-equilibration time; then retention, peak shape and selectivity can change especially where ionic interactions are prevalent.
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Is there anything special I need to do to my HPLC system to use Ascentis Express?
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Nothing special is required to use Ascentis Express HPLC columns. To obtain the full benefits of Ascentis Express, one should minimize dispersion or instrument bandwidth in the HPLC system (tubing, detector flow cell) as well as confirm the detector response system is set at a fast level. For more information, request Guidelines for Optimizing Systems for Ascentis Express Columns (T407102)
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How does the flow rate influence the water layer on the column?
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We are not aware of any systematic studies with respect to the impact of flow rate on HILIC separations. Our concern would be that as you move to higher flow rates, you might observe peak shape issues due to the slow kinetics of IEX and adsorption mechanisms. If the retention mechanisms for a given system are partition dominated, this should be of less concern. It will be a case by case cause and effect.
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