Features of Spursil™ Columns

• Combine high purity silica with unique polar modification technology
• Unique selectivity and enhanced resolution
• Silanol shielding for excellent peak shape
• Improved water wettability and stable retention in highly aqueous mobile phase conditions
• Excellent retention for polar compounds
• Extended range pH stability
• Choose from a variety of selectivities and hardware formats

Spursil™ polar-modified phases are based on ultra high-purity silica and novel polar modification technology. The unique bonded phases maximize polar retention and selectivity, while virtually eliminating silanol activity. The resulting polar-modified packing material contains a surface which is easily "wetted" with polar eluents and can run in highly aqueous conditions. The polar group also seems to play a role in base deactivation. We have seen some evidence that hydrogen bonding occurs between certain polar linkers and the silica surface, thereby decreasing the interaction of such silanols with basic components in the sample. Additionally, polar-modified phases often provide selectivity quite different from standard C18 phases.

Standard reverse phase columns, particularly C18 columns, often suffer from "phase collapse" and the retention of the compounds is severely diminished when used in combination with highly aqueous phases. This phenomenon is substantially reduced with Spursil™ because the polar modifications make the phase less hydrophobic and more wettable. Polar-modified phases remain fully extended in aqueous phases, allowing increased interaction between samples and the bonded phase. Spursil™ columns show good retention of polar compounds which tend to elute in the void volume on standard ODS phases.

Spursil™ columns are available in 3, 5, and 10 micron particle sizes, column lengths are ranging from 30 mm to 250 mm, and column dimensions are ranging from 2 mm to 21.2 mm. Ten micron preparative materials are available in bulk form.

Spursil™ Material Characteristics

High Efficiency

A chromatographic efficiency test is run on all columns. Plates per column and USP tailing factor are measured for each column to confirm packing efficiency. The USP tailing factor, a measure of peak symmetry at 5% of peak height, is a stringent indicator of peak shape. The test analytes are neutral hydrophobic compounds chromatographed in a acetonitrile: water mobile phase. The retention, selectivity, efficiency and peak symmetry of these molecules are measured to provide specifications for column performance.

Performance Test

Column: Listed on chromatograms   Dimension: 150 x 4.6 mm   Mobile Phase: MeCN:H2O = 60:40   Flow Rate: 1.0 mL/min   Temperature: 30 ºC   Detection: UV 254 nm   Sample: 1. Uracil 4. Toluene   2. Acetophenone 5. Naphthalene   3. Methyl benzoate

Batch-to-batch Reproducibility of Spursil™ C18

Reproducibility is imperative to any application in the laboratory. A new column may be interesting, but if the retentivity and selectivity are not consistent, it has little value in a validated method. Dikma has designed reproducibility into the Spursil™ family starting with tight specifications on the surface area, pore size and particle size of the silica. Bonding procedures are transferred to production through a manufacturing transfer validation procedure. All Spursil™ columns are manufactured at our ISO 9001 registered facility. This ISO 9001 registration provides quality oversight into all aspects of the manufacturing process, leading to a product that consistently meets exact specifications.

Reproducibility Test

Wide pH Range

One of the most important parameters in choosing a column is lifetime. Columns that last longer save time and money. Column failure often occurs because of hydrolytic instability of the stationary phase and silica as well as mechanical instability of the column packing (voiding) . The high phase coverage of all Spursil™ products plus proprietary endcapping creates a phase that is more stable against hydrolytic attack. Dikma's narrow specifications on surface area of the silica help to create a more consistent and well-packed column that minimizes shifting of the column bed during use. This reduces the chance of voiding. With Spursil™ columns, there is no worry about bonded phase hydrolysis, dissolution of the silica particle, or bonded phase collapse. The polar groups make the Spursil™ columns highly stable in hydrophilic conditions, from pH 1.5 to 10 and from 0 to 100% aqueous. All Spursil™ columns are tested at pH 1.5 and pH 10 under "real world" conditions with basic and acidic compounds to ensure good peak symmetry.

pH Stability Test

Column: Spursil™ 5 μm C18, 150 x 4.6 mm   Cat. No.: 82001   Mobile Phase: MeCN:20 mM phosphate buffer (pH 7) = 40:60   Flow Rate: 1.0 mL/min   Temperature: Ambient   Detection: UV 254 nm   Sample: 1. Uracil 3. Phenol   2. Pyridine 4. Benzene

Unique Selectivity

Spursil™ analytical HPLC columns are designed to provide enhanced retention and selectivity for highly polar compounds in the reversed-phase mode, as well as compatibility with highly aqueous mobile phases. These phases offer several advantages over traditional alkyl bonded phases including a unique selectivity, improved peak shape for basic compounds, and the ability to operate in highly aqueous mobile phases. These polar-modified columns give the same range of application but alternative selectivity compared to traditional alkyl bonded phases. Spursil™ phases provide additional charge density to adjacent silanol through electron delocalization, thereby removing mixed-mode interactions and improving peak symmetry. The best methods employ simple mobile phases. Spursil™ columns enable separation scientists who work with polar compounds to consistently deliver robust methods with good selectivity and excellent peak shape.

Cold Medicine

Column: Listed on chromatograms   Dimension: 150 x 4.6 mm   Mobile Phase: 0.1% TFA in MeCN:0.1% TFA in H2O = 35:65   Flow Rate: 1.0 mL/min   Temperature: Ambient   Detection: UV 220 nm   Sample: 1. Doxylamine 4. Dextromethorphan   2. Acetanilide 5. Diphenhydramine   3. Acetylsalicylic acid

Polar Acids*

Column: Listed on chromatograms   Dimension: 150 x 4.6 mm   Mobile Phase: MeOH:0.1% TFA in H2O = 20:80   Flow Rate: 1.0 mL/min   Temperature: Ambient   Detection: UV 210 nm   Sample: 1. Maleic acid 3. o-Toluamide   2. Procainamide 4. p-Hydroxybenzoic acid

Polar Compounds*

Column:  Listed on chromatograms   Dimension: 150 x 4.6 mm   Mobile Phase: 10 mM HCOONH4, pH 3   Flow Rate: 1.0 mL/min   Temperature: Ambient   Detection: UV 254 nm   Sample: 1. Thiourea 3. Adenine   2. Cytosine 4. Uridine

Outstanding Performance

Our Spursil™ family of reversed-phase columns, each prepared from the same high-purity silica, differ from one another solely and predictably in the hydrophobicity and polarity inherent in the stationary phase chemistry. The outstanding performance of Spursil™ columns allows users who are developing methods to achieve the right separation the first time, even with a simple mobile phase. This means faster development of HPLC methods, and more robust methods being transferred to QA / QC labs.

Anti-inflammatories

Column: Spursil™ 5 μm C18, 150 x 4.6 mm   Cat. No.: 82001   Mobile Phase: 0.1% HCOOH in MeCN:0.1% HCOOH in H2O = 55:45   Flow Rate: 1.0 mL/min   Temperature: Ambient   Detection: UV 254 nm   Sample: 1. Phenacetin 5. Flurbiprofen   2. Tolmetin 6. Diclofenac   3. Ketoprofen 7. Ibuprofen   4. Fenoprofen

Steroids

Column: Spursil™ 5 μm C18, 150 x 4.6 mm   Cat. No.: 82001   Mobile Phase: MeOH:H2O = 55:45   Flow Rate: 1.0 mL/min   Temperature: Ambient   Detection: UV 254 nm   Sample: 1. Prednisone 4. Dexamethasone   2. Cortisone 5. Hydrocortisone 21-acetate   3. Prednisolone 6. 11-α-Hydroxyprogesterone

Our Solution for Working Under Highly Aqueous Conditions

The separation of polar compounds under highly aqueous mobile phase conditions is not reproducible with conventional reversed-phase materials. A proprietary derivatization procedure enables Spursil™ columns to be penetrated by water without losing their hydrophobic property. This allows polar compounds to be separated with excellent peak shape and remarkable reproducibility. The separations of water-soluble vitamins and amino acids illustrate the features of the Spursil™ columns in improving selectivity and peak shape and optimizing retention.

Water-Soluble Vitamins

Column: Spursil™ 5 μm C18, 150 x 4.6 mm   Cat. No.: 82001   Mobile Phase: MeOH:10 mM K2HPO4 (pH 7) = 3:97   Flow Rate: 1.0 mL/min   Temperature: Ambient   Detection: UV 254 nm   Sample: 1. L-Ascorbic acid 5. Pyridoxol   2. Orotic acid 6. Nicotinamide   3. Pyridoxamine 7. Thiamine   4. Pyridoxal

Amino acids

Column: Spursil™ 5 μm C18, 150 x 4.6 mm   Cat. No.: 82001   Mobile Phase: 20 mM KH2PO4 (pH 2.5 by H3PO4)   Flow Rate: 1.0 mL/min   Temperature: Ambient   Detection: UV 210 nm   Sample: 1. Citrulline 3. Methionine   2. Valine 4. Tyrosine

Although they are designed for use in highly aqueous mobile phase conditions, Spursil™ columns are completely compatible with highly organic mobile phases. The ability to cover the full range of mobile phase composition, from 100% aqueous to 100% organic, is useful for developing gradient methods for analyzing complex samples. High phase density allows highly organic mobile phase for the best desolvation and MS sensitivity.

Tocopherols*

Column: Listed on chromatograms   Dimension: 150 x 4.6 mm   Mobile Phase: MeOH   Flow Rate: 1.0 mL/min   Temperature: 30 ºC   Detection: UV 295 nm   Sample: 1. δ -Tocopherol     2. γ -Tocopherol     3. α -Tocopherol

EasyGuard™ Columns

• Universal design to match any brand column
• A variety of optional bonded phase material
• Does not affect the analytical column resolution
• Long lifetime column cartridges, high performance and low price

Balance

Guard columns provide protection against contamination with minimal impact on column efficiency. The column diameter determines both the sample loadability and column efficiency. A small diameter will decrease the column lifetime, but a large diameter will adversely affect column resolution. EasyGuard™ columns effectively profect the analytical column without adversely affecting the resolution or column lifetime.

Simple to Use

• Flexibility to move PEEK fittings
• Matches any brand analytical column
• Low dead volume connection
• Rugged 316 stainless steel column holder

Easy-Lok™ Coupler ^New!

When the EasyGuard™ guard column connects to a 250 mm analytical column, it can not fit in certain ovens (such as Agilent 1200) because the length is not long enough. The Easy-Lok™ coupler resolves this issue by removing the pre-tightened connecting pipe by wrench before using.

HPLC Column Care Instructions

- Dropping or otherwise "shocking" columns can disrupt the column bed and cause peak splitting.
- Use of eluents in the pH range of 1-11 will maximize column life.
- The lower the operating pressure, the longer the operating life.

HPLC packings are subject to a rigorous array of QC tests in a ISO9001 compliant facility, with special emphasis on reagent purity, raw material traceability, and consistency in raw materials and finished products. A detailed analysis of all physical properties, chemical purity, chromatographic selectivity, and column packing efficiency is used to confirm that each lot of column is identical to all previous lots.

We know how important column consistency is to your work, so Dikma Technologies goes to great lengths to make certain that every HPLC column we ship to our customers is of the highest quality possible.

When you receive an HPLC Column from Dikma Technologies, please take the following simple steps:

(1) Check the column for signs of physical damage which may have occurred during shipping. Contact Dikma Technologies immediately to report any problems.
(2) Make sure that the column you received is the column that you ordered.
(3) Take note of the solvent contained in the column during shipping. (The solvent used for shipping is the same as that used as the mobile phase on the QC test chromatogram ). Before attempting to change solvents, make certain that the eluent you will be introducing into the column is COMPLETELY miscible with the eluent contained in the column, to avoid precipitation of buffer salts or other mobile phase additives.
(4) Test the column to verify column efficiency and back pressure (using one of the later-eluting components of the QC test sample). Contact Dikma Technologies immediately to report any problems.

There are several important things about your analytical method that will greatly affect column life and column performance, including sample preparation, solvent selection, and solvent preparation.

Sample Preparation
Non-ideal chemical and physical interaction of samples with the column frits and column packings is a primary source of problems. Most columns fail because the frit becomes clogged or the stationary phase at the head of the column becomes contaminated.

Samples should be filtered prior to injection. Even sample solutions which appear to be particulate-free can contain small solids which can clog the pores of the column inlet frit. Samples should be filtered through a 0.45µm or 0.2µm syringe filter before injection.

In addition to filtering samples, Guard cartridges can be used to trap "problems" before they reach the analytical column. Guard cartridges are essentially tiny HPLC columns that are cheaper to replace than the analytical column. Please contact Dikma Technologies of Ordering information for guard cartridges.

Solvent Selection
Columns last longest when they are used with benign eluents. Using eluents of high pH or low pH can dissolve silica or catalyze hydrolysis of the bonded phase. Try to stay within the pH range of 1 - 11 on the columns. If you must use a pH outside this range, column life might be reduced.

Solvent Preparation
Use a 0.45 µm or 0.2 µm filter for solvents as well as samples, even HPLC grade solvents!

COLUMN STORAGE
Column storage conditions can have a profound effect on column lifetime and performance-after-storage. Before extended storage (e.g. greater than 2 days), rinse the column COMPLETELY free of eluents containing buffers, ion-pair reagents, or inorganic solutes, by flushing with 20-50 column volumes of the eluent WITHOUT the dissolved additives. Then flush the column with 5-10 column volumes of water (reverse phase columns only). Then flush the column with 20 volumes of storage solvent (shown above)

In-Situ Column Cleaning
Columns that become fouled over time can sometimes be rejuvenated with an aggressive rinsing sequence, as shown below. In all cases, reverse the column (e.g. attach the outlet end of the column to the pump, and pump the eluent directly into a waste reservoir) and flush the column with 50ml volumes of the indicated solvents in the indicated sequences:

* If mobile phase contains a buffer, flush the column with the mobile phase MINUS the buffer first, to avoid precipitation of the buffer in the pure MeOH remaining in the column.