Yes. if possible you should always have at least a small amount of buffer in a HILIC system to help mediate/control IEX and other polar interactions that are bound to be present (even if at a low level). It is not so much the "buffering capacity" that is important, but the presence of the competing ions. We have found that their presence helps with day to day and column to column reproducibility. There are times when you need to eliminate the buffer, but aside from special circumstances, our recommendation is to include them.
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Ascentis® Express 90 Å HILIC (2.7 μm) HPLC Columns
L × I.D. 3 cm × 4.6 mm, HPLC Column
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About This Item
UNSPSC-Code:
41115700
eCl@ss:
32110501
NACRES:
SB.52
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Produktbezeichnung
Ascentis® Express HILIC HPLC Column, 2.7 μm particle size, L × I.D. 3 cm × 4.6 mm
Materialien
stainless steel column
Qualitätsniveau
Agentur
suitable for USP L3
Produktlinie
Ascentis®
Leistungsmerkmale
endcapped: no
Hersteller/Markenname
Ascentis®
Verpackung
1 ea of
Parameter
≤100 °C temp. range
600 bar max. pressure (9000 psi)
Methode(n)
HPLC: suitable
LC/MS: suitable
UHPLC-MS: suitable
UHPLC: suitable
L × ID
3 cm × 4.6 mm
Oberflächenbereich
135 m2/g
Verunreinigungen
<5 ppm metals
Matrix
Fused-Core particle platform
superficially porous particle
Aktive Matrixgruppe
silica phase
Partikelgröße
2.7 μm
Porengröße
90 Å
Betriebs-pH-Wert
1-8
Anwendung(en)
food and beverages
Trenntechnik
hydrophilic interaction (HILIC)
normal phase
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Allgemeine Beschreibung
Ascentis Express HPLC columns, through the use of Fused-Core® particle technology, can provide you with both the high speed and high efficiencies of sub-2 μm particles while maintaining lower backpressures. The combination of high efficiency and low backpressure benefits UPLC® (or other ultra high pressure system) users, as well as conventional HPLC users.
Visit the Ascentis Express home page for more information on this new column technology.
Visit the Ascentis Express home page for more information on this new column technology.
Rechtliche Hinweise
Ascentis is a registered trademark of Merck KGaA, Darmstadt, Germany
Fused-Core is a registered trademark of Advanced Materials Technology, Inc.
UPLC is a registered trademark of Waters
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Lagerklassenschlüssel
11 - Combustible Solids
WGK
WGK 3
Flammpunkt (°F)
Not applicable
Flammpunkt (°C)
Not applicable
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Paweł Kubica et al.
Journal of pharmaceutical and biomedical analysis, 127, 184-192 (2016-01-20)
Hydrophilic interaction liquid chromatography (HILIC) coupled with tandem mass spectrometry (MS/MS) was used to separate artificial and natural sweeteners approved for use in European Union (EU). Among three tested HILIC columns (BlueOrchid PAL-HILIC, Ascentis Express Si and Acclaim™ Trinity™ P2)
Tiziana Bertolini et al.
Journal of chromatography. A, 1365, 131-139 (2014-09-23)
A simple, sensitive and fast hydrophilic interaction liquid chromatography (HILIC) method using ultraviolet diode-array detector (UV-DAD)/electrospray ionization tandem mass spectrometry was developed for the automated high performance liquid chromatography (HPLC) determination of sodium risedronate (SR) and its degradation products in
Imran Ali et al.
Biomedical chromatography : BMC, 26(8), 1001-1008 (2012-01-13)
Superficially porous silica particles columns (SPSPCs) are manufactured by different companies. The most common have the brand names Halo, Ascentis Express and Kinetex. These columns provide super fast, sharp peaks and moderate sample loading and back pressure. These are available
Alexandre Grand-Guillaume Perrenoud et al.
Journal of chromatography. A, 1360, 275-287 (2014-08-19)
Superficially porous particles (SPP), or core shell particles, which consist of a non-porous silica core surrounded by a thin shell of porous silica, have gained popularity as a solid support for chromatography over the last decade. In the present study
Jan Soukup et al.
Journal of chromatography. A, 1374, 102-111 (2014-12-30)
Excess adsorption of water from aqueous acetonitrile mobile phases was investigated on 16 stationary phases using the frontal analysis method and coulometric Karl-Fischer titration. The stationary phases include silica gel and silica-bonded phases with different polarities, octadecyl and cholesterol, phenyl
Artikel
For separation of polar compounds including polar neutrals, polar acids, and polar and non-polar basic amines use our Ascentis® Express HILIC column.
Protokolle
We offer the tools for the analysis of the metabolites; including certified reference standards, enzymes, substrates, and chromatographic products.
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Would you advise addition of a buffer when using diol or amide stationary phase?
1 answer-
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Why is it recommended to run isocratically for HILIC methods?
1 answer-
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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Can peptide or protein samples be analyzed using HILIC columns?
1 answer-
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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Is there anything special I need to do to my HPLC system to use Ascentis Express?
1 answer-
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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In HILIC separations, what happens if the sample is an aqueous matrix? Does it always have a negative effect?
1 answer-
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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Do I need special fittings and tubing to connect Ascentis® Express HPLC Columns?
1 answer-
While operating pressures may not exceed the 400 bar (6,000 psi) capability of your traditional instruments, sustained pressures of about 200 bar (3,000 psi) will exceed the recommended pressure for conventional PEEK tubing and fittings at the column inlet. We recommend changing to stainless steel fittings in all high pressure locations and have designed special High Performance HPLC Fittings/Interconnects that will stay tight at pressures of 1,000 bar (15,000 psi) or greater, even when elevated column temperatures are employed.
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When you recommend only changing one parameter at a time, does this also refer to the total ionic strength?
1 answer-
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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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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How should I store the Ascentis Express HILIC column?
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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.
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