Features of Inspire™ C18 & C8 Columns

• Rapid separations with outstanding resolution
• Advanced bonding technologies
• High efficiency and outstanding lifetime
• Excellent separation characteristics over wide pH range
• Superior batch-to-batch reproducibility
• Choose from a variety of phases and hardware formats

Inspire™ columns are engineered with high purity raw silica, proprietary bonding techniques, tightly controlled manufacturing processes, and column packing procedures that provide today’s chromatographic laboratories with HPLC columns of unrivaled performance.

Inspire™ Material Characteristics

Superior Batch-to-batch Reproducibility
Reproducibility is essential for the selection of a HPLC column. Today’s chemists often need to establish new analytical methods to evaluate the latest pharmaceutical and biopharmaceutical products. The column they select has to provide the same chromatographic results over the entire lifespan of the new drug product. Chemists doing QA / QC also need a well-producible column, which ensures the accuracy of analytical results and high productivity of chromatographic laboratories. Inspire™ reverse phase columns undergo rigorous quality control testing to ensure long-term reproducibility, letting you increase your laboratory’s productivity and allowing for easier method transfer between labs.

Reproducibility Test

Five randomly selected batches demonstrated excellent reproducibility in the example shown:

Long Lifetime
Columns that last longer not only save you money, but also save you time in establishing and verifying methods for a new column. Inspire™ columns deliver guaranteed, consistent performance in optimizing the two key factors that control column lifetime: the packing material and the mechanical stability of the packed bed.

Lifetime Test     Column: Inspire™ 5 μm C18, 150 x 4.6 mm   Cat. No.: 81001   Mobile Phase: 0.1% TFA in MeCN:0.1% TFA in H2O = 30:70   Flow Rate: 1.0 mL/min   Temperature: Ambient   Detection: UV 220 nm   Sample: 1. Nadolol 5. Labetolol   2. Pindolol 6. Propranolol   3. Acebutolol 7. Alprenolol   4. Metoprolol

Stable from pH 1 - 11
Generally, the cause of shortened column lifetime relates to exposure under extreme pH mobile phases, which leads to hydrolysis of the bonded phase at low pH and dissolution of silica at high pH. The hydrolysis can also lead to significant changes in analyte retention time, making method reproducibility requirements difficult to achieve.
Dikma incorporates proprietary bonding and endcapping techniques, making Inspire™ packing much more stable across a broad pH range when compared to conventionally prepared material. Our packing platform also  effectively resists the typical ligand hydrolysis and silica dissolution seen with conventional columns. In both low pH and high pH tests, Inspire™ C18 columns undergo elution over 1,440 hours and show very little loss of retention time, capacity factor and symmetry, exhibiting their unsurpassed endurance and stability.

pH Stability Test

Column: Inspire™ 5 μm C18, 150 x 4.6 mm   Cat. No.: 81001   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

Efficient Method Development
Inspire™ columns provide chromatographic laboratories with advanced performance and robust, rugged methods that can be achieved across the entire log P range of -2 to 8, simplifying the method validation and transfer process. In the test below, aspartame, reserpine, cortisone and dioctyl phthalate are chosen to evaluate the Inspire™ columns with a sample representative of molecules encountered in drug discovery. The compounds vary in polarity (log P = -2 to 8) and molecular weight (MW 294 to 608). High quality separation of these components demonstrates the broad applicability of Inspire™ C18 to a range of compounds with drug-like properties.
Tang, L.; Fitch, W.L.; Alexander, M.S.; Dolan J.W. Anal. Chem., 2000, 72, 5211 - 5218.

LC / MS Performance Test Mix

Column: Inspire™ 5 μm C18   Dimension: 150 x 4.6 mm   Cat. No.: 81001   Mobile Phase: A: 0.05% HCOOH in MeCN     B: 0.05% HCOOH in H2O     Gradient: 10 - 90% A in 5 min, hold at 90% A for 5 min   Flow Rate: 1.0 mL/min   Temperature: Ambient   Detection: UV 220 nm   Sample: 1. Aspartame 3. Cortisone   2. Reserpine 4. Dioctyl phthalate

Engelhardt Test
The Engelhardt test is a stringent test we use to verify the selectivity of Inspire™ columns. It is based solely on stationary phase chemistry. In this test, aniline elutes before phenol and the three toluidine isomers coelute, indicating that the acidic silanol groups of Inspire™ columns are deactivated.

Engelhardt Test

Column: Inspire™ 5 μm C18   Dimension: 150 x 4.6 mm   Cat. No.: 81001   Mobile Phase: MeOH:H2O = 55:45   Flow Rate: 1.0 mL/min   Temperature: 30 ºC   Detection: UV 254 nm   Sample: 1. Thiourea 5. N,N-Dimethylaniline   2. Aniline 6. Ethyl benzoate   3. Phenol 7. Toluene   4. o,m,p -Toluidine 8. Ethylbenzene

Excellent Peak Shapes with Basic Molecules - but simple mobile phase*

Column: Inspire™ 5 μm C18, 150 x 4.6 mm   Cat. No.: 81001   Mobile Phase: A: 0.1% TFA in H2O     B: 0.1% TFA in MeCN     A:B = 60:40   Flow Rate: 1.0 mL/min   Temperature: Ambient   Detection: UV 254 nm   Sample: 1. Nordoxepin 5. Amitriptyline   2. Doxepin 6. Trimipramine   3. Desipramine 7. Clomipramine   4. Nortriptyline

Column: SunFire™ 5 μm C18, 150 x 4.6 mm   Mobile Phase: A: H2O     B: MeOH     C: 100 mM CH3COONH4, pH 6.0     A:B:C = 18:72:10   Flow Rate: 1.0 mL/min   Temperature: 30 ºC   Detection:  UV 254 nm   Sample: 1. Desipramine 4. Imipramine   2. Nortriptyline 5. Amitriptyline   3. Doxepin

Antihistamines at High pH*

Column: Listed on chromatograms   Dimension: 150 x 4.6 mm   Mobile Phase: MeOH:5 mM NH4HCO3 (pH 10) = 75:25   Flow Rate: 1.0 mL/min   Temperature: Ambient   Detection: UV 254 nm   Sample: 1. Maleic acid 4. Chlorpheniramine   2. Pheniramine 5. Brompheniramine   3. Doxylamine 6. Diphenhydramine

TCAs at High and Neutral pHs

Basic compounds tend to tail on alkyl phases because of the interaction with the silanols on the silica surface. This can often cause increased retention but loss in performance (peak shape). The most sensitive measurement of silanol interactions is achieved using highly basic probes with a pH 7 mobile phase. At this pH, many of the residual silanols are in their ionized form, and the basic probes are completely protonated. The protonated bases interact with the ionized silanols by an ion-exchange mechanism, and the degree of tailing is a direct measure of silanol activity. Inspire™ columns provide more symmetrical peaks and greater resolution for TCAs at high and neutral pHs, demonstrating their outstanding bonding and endcapping techniques.

TCAs at High pH*

Column: Listed on chromatograms   Dimension: 150 x 4.6 mm   Mobile Phase: MeOH:5 mM NH4HCO3 (pH 10) = 80:20   Flow Rate: 1.0 mL/min   Temperature: Ambient   Detection: UV 254 nm   Sample: 1. Nordoxepin 5.Imipramine   2. Doxepin 6.Amitriptyline   3. Desipramine 7.Trimipramine   4. Nortriptyline

TCAs at Neutral pH*

Column: Listed on chromatograms   Dimension: 150 x 4.6 mm   Mobile Phase: MeCN:20 mM phosphate buffer (pH 7) = 2:1   Flow Rate: 1.0 mL/min   Temperature: Ambient   Detection: UV 254 nm   Sample: 1. Desipramine 4. Amitriptyline   2. Nortriptyline 5. Trimipramine   3. Imipramine

Ultimate Performance

High phase density results in increased analyte-bonded phase interactions. These interactions impart greater selectivity and retention, leading to enhanced resolution.

Antibacterials*

Column: Listed on chromatograms   Dimension: 150 x 4.6 mm   Mobile Phase: 0.1% HCOOH in MeCN:0.1% HCOOH in H2O = 20:80   Flow Rate: 1.0 mL/min   Temperature: Ambient   Detection: UV 254 nm   Sample: 1. Sulfanilamide 6. Sulfamethoxypyridazine   2. Carbadox 7. Furazolidone   3. Sulfapyridine 8. Sulfamethoxazole   4. Sulfamerazine 9. Sulfisoxazole   5. Thiamphenicol 10. Oxolinic acid

Separation of Cephalosporin Antibiotics at Different Mobile Phase Conditions

Column: Inspire™ 5 μm C18, 150 x 4.6 mm   Cat. No.: 81001   Mobile Phase: MeOH:25 mM phosphate buffer (pH 3) = 20:80   Flow Rate: 1.0 mL/min   Temperature: Ambient   Detection: UV 230 nm   Sample: 1. Cefadroxil 5. Cefazoline   2. Ceftazidime 6. Cefoxitin   3. Cefaclor 7. Cefradine   4. Cephalexin

Mobile Phase: MeOH:100 mM acetate buffer = 20:80   Flow Rate: 1.0 mL/min   Detection: UV 254 nm   Sample: 1. Cefadroxil     2. Cefuroxime 4. Cefoxitin   3. Cefaclor 5. Cefradine

Mobile Phase: MeOH:0.1% TFA in H2O = 30:70   Flow Rate: 1.0 mL/min   Detection: UV 230 nm   Sample: 1. Ceftazidime 5. Cephalexin   2. Cefadroxil 6. Cefoxitin   3. Cefazoline 7. Cefradine   4. Cefaclor

Separation of TCAs and Benzos* Column: Listed on chromatograms   Dimension: 150 x 4.6 mm   Mobile Phase: 0.1% TFA in MeCN:0.1% TFA in H2O = 40:60   Flow Rate: 1.0 mL/min   Temperature: Ambient   Detection: UV 254 nm   Sample: 1. Nitrozepam 6. Triazolam   2. Nordoxepin 7. Nortriptyline   3. Alprazolam 8. Clonazepam   4. Diazepam 9. Trimipramine   5. Oxazepam

Organic Acids*

Column: Listed on chromatograms   Dimension: 150 x 4.6 mm   Mobile Phase: 25 mM KH2PO4 pH 2.5   Flow Rate: 1.0 mL/min   Temperature: Ambient   Detection: UV 210 nm   Sample: 1. Oxalic acid 5. Acetic acid   2. Tartaric acid 6. Citric acid   3. Malic acid  7. Fumaric acid   4. Lactic acid 8. Succinic acid

Flavonoids

Column: Listed on chromatograms   Dimension: 150 x 4.6 mm   Mobile Phase: MeCN:0.085% H3PO4 = 20:80   Flow Rate: 1.0 mL/min   Temperature: Ambient   Detection: UV 280 nm   Sample: 1. Gallic acid 5. p-Coumaric acid   2. Catechin 6. Quercitrin   3. Caffeic acid 7. Myricetin   4. Vanillic acid

Herbicides

Column: Listed on chromatograms   Dimension: 150 x 4.6 mm   Mobile Phase: 0.1% TFA in MeCN:0.1% TFA in H2O = 40:60   Flow Rate: 2.0 mL/min   Temperature: Ambient   Detection: UV 214 nm   Sample: 1. Dalapon     2. 2,4-D     3. 2,4,5-T

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.