Microsolv make Cogent HPLC Column including a range of specialties
MicroSolv Technology Corporation is a pioneer in manufacture of Silica hydride based Cogent HPLC columns. Indeed, their Silica hydride columns and ANP methods are the choice for separating highly polar small molecules. Besides, they are used worldwide as favoured product in metabolomics industry
MicroSolv Technology Corporation is designing, producing, and selling specialty products for the separation and purification sciences. Their product range includes HPLC and GC columns, vials, and tools. The purpose of most products are to reduce workload or to increase effectiveness in chromatography.
We represent MicroSolv Technology since about 20 years in the German speaking countries.
MicroSolv is in 9158 Industrial Boulevard NE, Leland, NC 28451, USA.
Speaking about this, Silicas come in three different types
Silica Type A: is Silica gel manufactured prior 2000. Silica gel caries four different silanol groups on the particle surface. Indeed, the ratio of acidity and reactivity of silanol groups varies. In the early era it was difficult to control the ratios of the various silica moieties. Conjugating organic ligands to the surface was a gamble.
Silica Type B; After 2000 the industry launched ultrapure base deactivated Silica gel. Silica gel B type provides sharp peaks and good reproducibility with stabile methods.
Cogent type B Silica specialty products are:
Chiral Columns: Cogent EE, Cellulose Coated
High Performance Reverse phase columns: Cogent hQ,
Longer Chain, Hydrophobic Columns: Cogent C27, Cogent C30
Silica Type C:
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Silica hydride has Si-H moieties on its surface. Conjugating organic ligands form strong Si-C bonds (like C-C bonds]
The surface of the TYPE-C™ columns are populated with silica hydride groups instead of silanols. Thus the particles are slightly hydrophobic. As a consequence, it will adsorb and desorb solvents much more easily than ordinary silica. Type C™ silica-hydride columns are bonded with the same ligands that are used with silica gel columns.
The following Type C Cogent columns are available: Cogent Bidentate C8, C18 and UDC-Cholesterol columns.
Benefits of Cogent HPLC Silica hydride Technology from Microsolv
Benefit 1 : Faster turn-around-time between runs. Most Type B HPLC columns require 15-20 columns volumes to equilibrate. TYPE-C columns only require 1-2 column volumes. You save time, solvents and cost are lower per analysis.
Benefit 2 : Easy method development. Retain both polar and non polar compounds on the same column. In less than 60 minutes you arrive at a optimised method. Most methods developed on TYPE-C columns use the same mobile phase even when changing to different bonded phases. They are reliable and reproducible HILIC columns Read under HILIC
Benefit 3 : Extended column lifetime, less lifetime failures and investigations. Direct silicon carbon bonds and the lack of a need for end capping makes these columns very robust. Indeed, they last 10-15 times longer than columns based on older silica technology.
Benefit 4 : Selectivity enhancements. All Type C column phases can be used in any of the (3) modes of chromatography. Reverse Phase, Normal Phase or Aqueous Normal phase. Also, one can switching between modes without hysteresis or damaging the column.
Benefit 5 : Easy to use. Simple singular mobile phases such as acetonitrile and water with acid or base can usually separate most compounds.
Benefit 6 : Save time and money, do more with less. The initial cost of column acquisition is similar to other market leaders. Such as reliability, precision, fast equilibration, savings in personnel and instrument time. Solvents, extended column lifetime and low column failure rate, gives incredible overall column life-time savings.
Eluent selection strategy for Microsolv Cogent HPLC
- A wide range of solvents can be used with Cogent TYPE-C Silica™ Based HPLC Columns..
- Selecting which TYPE-C column to use (C18, C8, Cholesterol or Silica-C) will be determined by your compounds of interest. Highly polar mixtures without non polar compounds are ideal for Silica-C Chromatography. Use Bidentate C8, C18 or UDC Cholesterol for compounds with both polar and non polar components.
Step 1: After you have properly installed and conditioned the column start with a typical reverse phase gradient run. We suggest starting with an acidified mobile phase of Water as component A and Acetonitrile as component B. Acidify both components with up to 0.5% of an acid. You may use Formic, Methyl Phosphonic or Acetic Acid. If you are not using LC-MS, TFA (up to 0.1%) is another good candidate.
Step 2: Run about 6 column volumes of the mobile phase in Step 1 at 95% Water.
- Step 3: Set up your instrument to run a shallow gradient from 95% Water (A) to 40% Water (A) over 20 minutes on a 75mm long column. For longer or shorter columns, increase the gradient time proportionally.This long and shallow gradient will be very beneficial for determining the optimal gradient or isocratic method to run your mixture with later. For sharper peaks and less retention, run a shorter (Steeper) gradient from the same starting points to end points.
- Step 4: Equilibrate the column by running 100% Acetonitrile for approximately 2 minutes for the 75mm long column.
- Step 5:. Set up your instrument to run a shallow gradient using the same mobile phase to run from 90% Acetonitrile (B) to 40% Acetonitrile(B) over 20 minutes for a 75mm long column. For longer columns increase the gradient time proportionally. This long and shallow gradient will be very beneficial for determining the optimal gradient or isocratic method to run your mixture with later. For sharper peaks and less retention, run a shorter (Steeper) gradient from the same starting points to end points.
- Step 6: Evaluate both gradient runs for retention time, peak shape and elution order. Since analyte retention on these columns is compound and method specific some compounds may not retain in Step 3 (Reverse Phase) and some may not retain in Step 5 (Aqueous Normal Phase). However, one column could produce an isocratic run which retains both polar and non polar compounds.
Cogent HPLC Column selection
- Note: Cogent Bidentate C8, C18 and UDC-Cholesterol columns have a unique quality in that they may retain polar compounds not retained on other columns while run at 100% Water without loss of retention with continued use. You could insert an isocratic run at 100% acidified Water after Step 3 and before Step 4.
- If you do not find satisfactory results or once you have established some retention and selectivity, you need to optimize the method for your target compounds. We have listed some suggestions below that will offer some ideas. If you are not satisfied with the results, please contact our technical support team.
For some organic acids that do not retain at low pH in an ANP mobile phase, you can try a high pH ANP mobile phase for retention. A rule of thumb is that for acids you would want to be from 1-2 pH units above the pKa. Some examples of additives that may work for you are Ammonium Acetate, Ammonia, Sodium Acetate, Ammonium Formate or Sodium Formate.
- The choice of solvents that you make for ANP will have different resolving powers for Acids and Bases. Acetone, Acetonitrile, THF, Ethanol, Acetonitrile/Methanol, Methanol and then Water will have a descending strength of retention in ANP.
All Cogent TYPE-C columns have the ability to perform in ANP. Depending on your sample mixture and matrix. Different columns will produce different peak shapes, retention and selectivity. Column hydrophobicity increases in the following order: Silica-C™, Bidentate C8, UDC-Cholesterol and Bidentate C18,
Equilibration between gradient runs:
- Although the Cogent TYPE-C Columns equilibrate quickly, the mobile phase solvents you are using will determine how much equilibration time you may need between runs. The hydride surfaces have a preference for Methanol. Therefore, it may take longer to equilibrate if methanol is used in a gradient end and not in the beginning. To equilibrate, use the starting point of your gradient as the solvent composition for equilibration.
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