Typically when we refer to pH in HILIC, we use the effective pH or the pH as measured after the addition of organic. The point is that we should always define what pH we are stating. The common way to distinguish is using notation of w/w pH or s/w pH (usually superscript/subscript). The notations mean superscript = solvent the pH is measured in (s would indicate some mixture of aqueous:organic) and the subscript = the solvent the pH meter is calibrated in (typically water (or w) as we readily have calibration standards).
53946-U
Ascentis® Express 90 Å HILIC (2.7 μm) HPLC Columns
L × I.D. 15 cm × 2.1 mm, HPLC Column
Synonym(s):
Core-shell (SPP) Fused Core Si HPLC column
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1 EA
$813.70
About This Item
UNSPSC Code:
41115700
eCl@ss:
32110501
NACRES:
SB.52
$813.70
List Price$1,030.00Save 21%In StockDetails
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Product Name
Ascentis® Express HILIC, 2.7 μm HPLC Column, 2.7 μm particle size, L × I.D. 15 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.
15 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
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.
Legal Information
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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Pricing
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Storage Class
11 - Combustible Solids
wgk_germany
WGK 3
flash_point_f
Not applicable
flash_point_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
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
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
Articles
For separation of polar compounds including polar neutrals, polar acids, and polar and non-polar basic amines use our Ascentis® Express HILIC column.
Protocols
We offer the tools for the analysis of the metabolites; including certified reference standards, enzymes, substrates, and chromatographic products.
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When referring to the pH of the mobile phase (pH 3, pH 4, etc.), does that refer to the aqueous part of the mobile phase?
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Can I use Ascentis Express on a UHPLC system?
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Yes. Ascentis Express columns are packed in a way making them suitable for these ultra high pressure instruments. In fact, Ascentis Express outperforms sub-2 μm micron columns on many applications since Ascentis Express provides the benefits of sub-2 μm particles but at much lower back pressure. These benefits include the capability of providing fast HPLC and higher resolution chromatography. The Fused-Core particle consists of a 1.7 μm solid core and a 0.5 μm porous shell. A major benefit of the Fused-Core particle is the small diffusion path (0.5 μm) compared to conventional fully porous particles. The shorter diffusion path reduces axial dispersion of solutes and minimizes peak broadening.
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What column do you recommend for an anionic compound?
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If the acids are hydrophilic or you can adjust the pH to make them hydrophilic enough, any of the phases that exhibit HILIC partitioning are possible (bare silica, OH5, diol, Zwitterionic, amide). We typically go with the OH5 first to try and avoid any negative impacts on the like charge.
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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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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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How can I measure my instrument bandwidth (IBW) and determine what Ascentis® Express HPLC Columns can be used with minimal efficiency loss created by too much internal instrument volume?
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The Guide to Dispersion Measurement has simple instructions on how to measure IBW and can be found at sigma-aldrich.com/express.
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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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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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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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Would you advise addition of a buffer when using diol or amide stationary phase?
1 answer-
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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