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Superior Performance and Reliability
Enduro GC and HPLC Columns
SGE has five decades of expertise in Chromatography providing unique understanding of the customer’s requirements enabling us to optimize column chemistry for analyte separations. The Enduro range of gas and liquid chromatography columns provide an excellent resource for routine analysis in a wide variety of fields including environmental, food, petrochemical, forensics and pharmaceutical.
Enduro GC and LC Columns2
GC Capillary Column SelectionSelecting the appropriate capillary column for your separation requires you to consider four fundamental parameters – phase type, film thickness, column ID and column length. The following serves as a guide to help you choose the right column for your work.
1. Phase Selection
• Select the least polar phase that will perform the separation you require (EN1, EN5, EN5MS and EN8).
• Non-polar stationary phases separate analytes predominantly by order of boiling point. By increasing the amount of phenyl and/or cyanopropyl content in the phase, and the separation is then influenced more by differences in dipole moments or charge distributions (EN10 (1701), ENX50 and ENX70).
• To separate compounds that differ more
in their hydrogen bonding capacities (for example aldehydes and alcohols), polyethylene glycol type phases are best suited - EN20 (WAX) and ENSGW.
2. Internal Diameter
• The smaller the diameter the greater the efficiency, leading to improved resolution. Fast columns, those with a 0.1 mm ID, are used because the same resolution can be achieved in a shorter time.
3. Film Thickness
• For samples with a variation in solute concentration, a column with film thickness >0.25 µm is recommended. This will reduce the possibility of broad overloaded peaks co-eluting with other compounds of interest. If the separation of two solutes is sufficient and co-elution is still unlikely, even with large differences in concentration, then a thinner film can be used.
• The greater the film thickness the greater the retention of solutes, therefore the higher the elution temperature.
• From the phase ratio value, a column can be categorized for the type of application it would best suit:
• The smaller the phase ratio value, the greater the ratio of phase to the column inner diameter, making it better suited for analyzing volatile compounds.
• Conversely, columns that have thin films are generally better suited for high molecular weight compounds and are characterized by large phase ratio values.
• Maintain phase ratio among different ID columns to yield similar chromatography.
Gas Chromatography Capillary Column Range
The Enduro range of stationary phases consist of basic polymer units with functionalities that can be modified by the addition of various moieties during synthesis. These moieties can be added in various amounts to create different concentrations of a particular functionality.
3Enduro GC and LC Columns
4. Column Length
• Always try to select the shortest column length that will provide the required resolution for the application. If the maximum column length available is being used and resolution of the sample mixture is still inadequate then try changing the stationary phase or internal diameter.
• Resolution is proportional to the square root of the column efficiency; therefore, doubling the column length will only increase the resolving power of the column by approximately 40%.
Table 1. Phase ratios available for the Enduro range of GC capillary columns. Maintaining a similar phase ratio when changing column internal diameters will ensure that your chromatographic parameters will not require substantial changes.
Application Range for Varying Phase Ratios16-100 Gases, Low Molecular Weight Hydrocarbons, Solvents, Volatile Halogens
(M.W.16-250)100-320 Semi-volatiles, General Applications (M.W. 100-700)320-1325 High Molecular Weight Hydrocarbons, Waxes, Petroleum Products (M.W.
300-1500)
Film Thickness (μm)
Column ID (μm)100 220 250 320 530
0.1 250
0.25 220 250 320
0.5 110 125 160
1 80 132
3 44
Enduro GC and LC Columns4
Enduro Phase Description and Recommended Applications
Enduro Phase Description Recommended Applications
EN1 100% Dimethyl Polysiloxane Suitable for analysis of hydrocarbons, aromatics, pesticides, phenol, herbicides, amines.
EN1MS SolGel + 100% Dimethyl Polysiloxane
Recommended for highly active compounds.
EN5 5% Phenyl Polysiloxane General purpose, aromatics, pesticides, herbicides, drugs of abuse, hydrocarbons, solvent impurities, PCB congeners or Aroclor mixes, essential oils, semivolatiles.
ENX5 5% Phenyl Polysilphenylene-siloxane
Ultra trace analyses, pesticides/herbicides, hydrocarbons, solvents, phenols, amines, GC/MS and other specific detector applications.
EN8 8% Phenyl Polycarboranesiloxane PCB congener analyses, nitro-substituted aromatics, polynuclear aromatic hydrocarbons, pesticides/herbicides.
EN10 (1701) 14% Cyanopropylphenyl Polysiloxane
Environmental analyses (EPA methods 608 and 8081), pesticides/herbicides, drugs of abuse, pharmaceuticals.
EN20 Bondable Polyethylene Glycol Alcohol, free acids, fatty acid methyl esters (FAMEs), aromatics, solvents, essential oils.
ENX50 50% Phenyl Polysilphenylenesiloxane
EPA methods 604, 608, 8060, 8081, triazines/herbicides, drug screening, steroids and a variety of pharmaceutical applications GC2D
ENX70 70% Cyanopropyl Polysilphenylenesiloxane
Fatty acid methyl esters (FAMEs), carbohydrates, pharmaceuticals, GC/MS applications
ENSGW SolGel + Polyethylene Glycol Recommended for highly active compounds. Alcohol, free acids, fatty acid methyl esters (FAMEs), aromatics, solvents, essential oils.
EN624 Cyanopropylphenyl Polysiloxane EPA method 624, drinking water volatiles, chlorinated hydrocarbons, solvents, Excellent for U.S. EPA Methods
5Enduro GC and LC Columns
Minimum TemperatureMaximum Continuous
Operating TemperatureMaximum Cycling
TemperatureThe temperature below which the capillary column will not separate components due to loss of partitioning in the stationary phase.
The maximum temperature at which a capillary column can be held for 72 hours with no significant change. Enduro capillary columns are designed to pass all criteria measured by their test analysis after 72 hours at their Maximum Continuous Operating Temperature
The maximum cycling temperature to which a capillary column can be taken for short periods (up to 30 minutes) without causing serious bleed problems or degradation of the phase. This is usually higher than the Maximum Continuous Operating Temperature. The lifetime of a capillary column is affected by the amount of time it spends at high temperatures.
Enduro GC Capillary Column Operating Temperature
For each Enduro GC column phases temperature limits are represented three ways:
Maximum Continuous Operating Temperature
Minimum Temperature
-60 to 320 / 340 Maximum Cycling Temperature
Enduro GC and LC Columns6
EN1 - 100% dimethyl polysiloxane; classic crosslinked dimethyl polysiloxane technology.
• Excellent general purpose non-polar GC column.• Suitable for all routine analyses.• 320 – 340 °C upper temperature limit – dependent on film thickness.
ID (mm) Film Thickness (μm)
Length (m) Temperature Limits (°C)
Part No.
0.22 0.25 30 -60 to 320/340 053101
0.25 0.25 30 -60 to 320/340 053120
0.25 0.25 60 -60 to 320/340 053121
0.32 0.25 30 -60 to 320/340 053107
0.32 1.0 60 -60 to 320/340 053109
0.53 1.0 30 -60 to 320/340 053119
EN1MS - 100% dimethyl polysiloxane in a Sol-Gel Matrix
• Same selectivity as EN1.• Improved inertness.• Low bleed for MS applications.
ID (mm) Film Thickness (μm)
Length (m) Temperature Limits (°C)
Part No.
0.25 0.25 30 0 to 340/360 053280
0.32 0.25 30 0 to 340/360 053281
EN5 - 5% phenyl / 95% dimethyl polysiloxane
• Ideal general purpose non-polar column.• Excellent inertness.• Good thermal stability.
ID (mm) Film Thickness (μm)
Length (m) Temperature Limits (°C)
Part No.
0.22 0.25 30 0 to 340/360 053131
0.25 0.25 30 -60 to 320/340 053139
0.32 0.25 30 -60 to 320/340 053137
ENDURO GC Capillary Columns
7Enduro GC and LC Columns
ENX5 - 5% phenyl polysilphenylene-siloxane
• General purpose GC column – suitable for over 80% of all routine analyses performed by gas chromatography.
• Excellent inertness, suitable for trace analysis.• Superior thermal stability, suitable for high temperature GCMS applications.
ID (mm) Film Thickness (μm)
Length (m) Temperature Limits (°C)
Part No.
0.1 0.1 10 -40 to 360/370 053220
0.25 0.25 30 -40 to 360/370 053221
0.25 0.25 60 -40 to 360/370 053222
0.32 0.25 30 -40 to 360/370 053223
0.32 0.25 60 -40 to 360/370 053224
EN8 - 8% phenyl polycarborane-siloxane
• Preferred column for polychlorinated biphenyl (PCB) compounds.• Unique high temperature phase suited for the analysis of persistent organic pollutants
(POPs).• Excellent thermal stability, suitable for GCMS.
ID (mm) Film Thickness (μm)
Length (m) Temperature Limits (°C)
Part No.
0.25 0.25 30 -20 to 360/370 053250
0.25 0.25 60 -20 to 360/370 053251
EN10 - 14% cyanopropylphenyl polysiloxane
• Used for organochlorine pesticides analysis.• Ideal for EPA methods 608 and 8081.
ID (mm) Film Thickness (μm)
Length (m) Temperature Limits (°C)
Part No.
0.22 0.25 30 35 to 240/250 053151
0.25 0.25 30 35 to 240/250 053164
0.32 0.25 30 35 to 240/250 053157
0.53 1.0 30 35 to 240/250 053163
Enduro GC and LC Columns8
EN20 - bondable polyethylene glycol (Wax)
• Industry standard wax column.• Low bleed.• Polar phase suitable for hydrogen bonding analytes.
ID (mm) Film Thickness (μm)
Length (m) Temperature Limits (°C)
Part No.
0.22 0.25 30 20 to 260/280 053181
0.22 0.25 60 20 to 260/280 053182
0.25 0.25 30 20 to 260/280 053197
0.25 0.25 60 20 to 260/280 053199
0.25 0.50 30 20 to 260/280 053198
0.32 0.50 30 20 to 260/280 053187
0.32 0.50 60 20 to 260/280 053188
0.53 1.0 30 20 to 260/280 053196
ENSGW - bondable polyethylene glycol (PEG) in a Sol-Gel matrix
• Excellent high temperature wax phase.• Bonded polyethylene glycol.• Less susceptible to damage by oxygen than conventional wax phases.
ID (mm) Film Thickness (μm)
Length (m) Temperature Limits (°C)
Part No.
0.25 0.25 30 30 to 260/280 053290
0.32 0.50 30 30 to 260/280 053291
ENX50 - 50% phenyl polysilphenylene-siloxane
• Mid polarity, industry standard for 2D GC second dimension.• High temperature stable, low bleed.• Ideal for a range of EPA methods and pharmaceutical applications.
ID (mm) Film Thickness (μm)
Length (m) Temperature Limits (°C)
Part No.
0.10 0.10 10 10 to 330/350 053230
0.25 0.25 30 10 to 330/350 053231
9Enduro GC and LC Columns
Example GC applications
Polynuclear Aromatic Hydrocarbons (PAH) Analysis on ENX50
ENX70 - 70% cyanopropyl polysilphenylene-siloxane
• Custom designed for separation of Fatty Acid Methyl Esters (FAMEs).• Industry standard column for FAME analysis.• Temperature stable polar phase.
ID (mm) Film Thickness (μm)
Length (m) Temperature Limits (°C)
Part No.
0.25 0.25 30 50 to 250/260 053240
0.25 0.25 60 50 to 250/260 053241
0.32 0.25 30 50 to 250/260 053242
EN624 – Cyanopropylphenyl polysiloxane
• US EPA 624 optimized column.• Designed for volatiles analysis.• Ideal for EPA624, SW-846 methods 8240/8260 and USP G43 method.
ID (mm) Film Thickness (μm)
Length (m) Temperature Limits (°C)
Part No.
0.53 3.0 30 0 to 230/240 053210
0.53 3.0 60 0 to 230/240 053211
Column: ENX50 30 m x 0.25 mm ID x 0.25 μm filmPAH standard: 10 ng/ μL in dichloromethaneDetector: MSD
Components
1. Naphthalene2. Acenaphthylene3. Acenaphthene4. Fluorene5. Phenanthrene6. Anthracene7. Fluoranthene
8. Pyrene9. Benzo (a) anthracene10. Chrysene11. Benzo (b) fluoranthene12. Benzo (k) fluoranthene13. Benzo (a) pyrene14. Indeno (1,2,3-c,d) pyrene15. Dibenzo (a,h) anthracene16. Benzo (g,h,i) perylene
Enduro GC and LC Columns10
8141 Organophosphorous Pesticide Mix on ENX5
Column: ENX5 30 m x 0.25 mm ID x 0.25 μm film8141 Standard: 10 ng/ μL in dichloromethaneDetector: MSD
Analysis of Omega - 3 Fatty Acids using a Highly Selective GC Capillary Column
Column: ENX70; 30 m x 0.25 mm ID x 0.25 μm filmFAME standard
Components
1. 4-Chloro-3-nitrobenzotrifluoride2. Dichlorvos3. 1-Bromo-2-nitrobenzene4. α-Mevinphos5. Tri-butylphosphate6. Ethoprop7. Sulfotepp
8. Naled9. Phorate10. Demeton11. Diazinon12. Disulfoton13. Methyl parathion14. Ronnel15. Chlorpyrifos16. Fenthion
17. Trichlorinate18. Tetrachlorvinphos19. Tokuthion20. Impurity21. Fensulfothion22. Impurity23. Triphenylphosphate24. Guthion25. Coumaphos
Components:
1. C62. C83. C104. C115. C126. C137. C148. C14:1 cis 99. C1510. C1611. C16:1 cis 9
12. C1713. C17:1 cis 1014. C1815. C18:1 trans 916. C18:1 cis 917. C18:1 cis 1218. C18:2 trans 9, 1219. C18:2 cis 9, 1220. C18:3 cis 6, 9, 1221. C18:3 cis 9, 12, 1522. C18:4 Cis 6, 9, 12, 1523. C20
24. C21:1 cis 1125. C20:2 cis 11,1426. C20:3 cis 8, 11,1427. C20:4 cis 5, 8, 11, 1428. C20:3 cis 11, 14, 1729. C2230. C20:5 cis 5, 8, 11, 14, 1731. C22:1 cis 1332. C22:4 cis 7, 10, 13, 1633. C2434. C22:6 cis 4,7,10,13,16,19
55 10 15 20 25 30 35min
pA
45
40
35
30
25
20
15
10
5
1
2
3
4
5
6
7
89
1011
13
12
14
15
16
17
1819 20
21
22
23 24
25
2627
28
3129
30
32
33
34
11Enduro GC and LC Columns
Analysis of 18 Alcohols on EN20
Column: EN20 30 m x 0.25 mm ID x 0.25 μm filmDetector: FID
Analysis of Various Monomers on ENSGW
Column: ENSGW 30 m x 0.25 mm ID x 0.25 μm filmSample: 250 ppm in HexaneDetector: FID
Components
1. Methanol2. Propan-2-ol3. Ethanol4. sec-Butan-1-ol5. n-Propanol + 2-Methyl-3-Buten-2-ol6. d,l-3-Methyl-2-Butan-1-ol7. Pentan-3-ol8. d,l-2-Pentan-1-ol9. n-Butanol10. 2,4-Dimethyl, Pentan-3-ol11. Hexan-3-ol12. 2-Methyl Prop-2-en-1-ol13. Crotyl Alcohol (2-Buten-1-ol)14. Hexan-2-ol15. Pentan-1-ol16. 2-Methyl Pentan-1-ol17. 2-Ethyl Butan-1-ol18. Hexan-1-ol
Components
1. Ethyl acrylate2. n-Butyl acrylate3. n-Butyl methacrylate4. Styrene5. 2-Ethylhexyl acrylate
6. Acrylic acid7. Hydroxypropyl acrylate8. Methacrylic acid9. Unknown10. Hydroxyethyl acrylate11. Lauryl Methacrylate
Enduro GC and LC Columns12
The Enduro range of C18 reversed phase HPLC columns are ideal for routine analysis of small molecules in pharmaceutical, environmental and food analysis. The high carbon loading of this phase results in the excellent retention of very polar compounds even in 100% aqueous conditions.
• Each Enduro column is individually tested with a guaranteed column efficiency of ≥ 90,000 theoretical plates.
• Improved peak symmetry – ultra high purity silica, highly end-capped.• Superb pH stability over a pH range from pH 1.0 - 9.0.• High loading capacity – specific surface area: 300 m²/g.
Are all C18 columns the same?
There are many C18 column brands available so how do you compare one C18 silica to the other?A number of test procedures have been published for the evaluation of C18 column performances. One such test was developed by the National Institute for Standardisation (NIST) and the results of this test for a number of commercially available columns, is published by the United States Pharmacopeia.The NIST SRM 870 test evaluates the silica’s properties such as metal activity, silanol activity and selectivity. Enduro C18 demonstrates superior performance with respect to minimal metal activity (Quinazrin peak shape) and free silanol activity (Amitriptyline peak shape).
Ultra-High Purity Silica and Bonded Phase – the SRM 870 Test
Enduro Liquid Chromatography Column Range
Silica Brand X ODS 5 micronSilica Brand Y ODS 5 micronSGE Enduro
Time (min)
Abs
(25
4 nm
)
250000
200000
150000
100000
50000
00 2 4 6 8 10 12 14 16 18 20
13Enduro GC and LC Columns
Enduro (4.6 mm x 150 mm) alkaline pH stability in 0.025 M sodium tetra borate solution and 40 °C – 30 injections of the column text mix with an overall eluted volume of 6,000 mls
0 2 4 6 8 10
100000
90000
80000
70000
60000
50000
40000
30000
20000
10000
0
Abs
(25
4 nm
)
Time (min)
2nd Injection - 50 mL
10th Injection - 500 mL
20th Injection - 1000 mL
30th Injection - 1500 mL
Enduro (4.6 mm x 150 mm) acid pH stability in 1 % TFA and 40 °C – 30 injections of the column test mix with an overall eluted volume of 6,000 mls
Time (min)
Abs
(25
4 nm
)
120000
100000
80000
60000
40000
20000
0
0 1 2 3 4 5 6 7 8 9 10
2nd Injection - 50 mL
10th Injection - 500 mL
20th Injection - 1000 mL
30th Injection - 1500 mL
How robust is Enduro to variation in pH of the mobile phase?
Enduro columns demonstrate minimal impact of acid and alkali pH on the retention time after the 1st or after the 30th injection. Column was exposed to pH 1.0 and pH 9.0 buffers at 40 °C and a test mix of theophilline, 4-nitro aniline, phenetole, methyl benzoate and xylene injected every 200 mls ~25 column volumes - shown in the figures below.
Enduro GC and LC Columns14
What if I need a C18 column that can withstand 100% aqueous conditions – is the Enduro C18 column sufficiently robust to withstand these conditions?
The best way to answer this question is to review the separation of fruit acid standards as well as a sample of Cranberry juice. In this application a stop-flow experiment demonstrates the column’s stability in 100% aqueous conditions and there is no evidence of phase collapse. After extended exposure to the analysis conditions, the column shows sharp symmetrical peaks and a good resolution for all six organic acids.
3 Quinic
4 Malic
6 Citric
1 Oxalic
2 Tartaric
5 Ascorbic
Column: 250 x 4.6 mm Enduro C18 (5 µm)Mobile Phase: 20 mM KH2PO4, pH 2.5 (H3PO4) 1 mL/minTemperature: 30 °CDetection: 210 nmSamples: i) Test Mix (Organic Acids)
ii) Cranberry Juice
15Enduro GC and LC Columns
I need a C18 carbon column with a high carbon load to retain polar compounds. How selective is Enduro C18 for polar compounds?
To address this question we examined the retention of artificial sweeteners and benzoate in soft drink. The purpose of the test was two-fold. First to characterise the retentive behaviour of the column by the retention of the benzoic acid and second to quantitate silica matrix effects by the peak shape of aspartame. Aspartame is a sensitive probe for non-specific interactions with the silica matrix. The chromatogram shows a good retention for benzoic acid (k’=9) highlighting the high hydrophobicity of the phase – Enduro is an excellent choice for polar compounds considering its stability in 100% aqueous mobile phase. The good peak shape of the aspartame peak demonstrates the high quality of the stationary phase with very little evidence of non-specific interactions visible.
2 Acesulfame
1 Saccharin
4 Benzoate
3 Aspartame
Column: 150 x 4.6 mm Enduro C18 (5 µm)Mobile Phase: 0.5% H3PO4 - AcN 80:20, 2 mL/minTemperature: AmbientDetection: 210 nm 220 nm
254 nm
SGE Analytical Science would like to thank Dr. Constantin S. Sychov from the “SCAN Chromatography Training Center” in Orel, Russia for performing the tests and providing the chromatograms.
Enduro GC and LC Columns16
0 1 2 3 4 5 6 7 8 9 10
Time (min)
2500th Injection (1185 psi)
2000th Injection (1173 psi)
1500th Injection (1162 psi)
1000th Injection (1150 psi)
500th Injection (1150 psi)
1st Injection (1144 psi)
Reproducibility of 2,500 injections of the column test mix on an Enduro C18 150 mm x 4.6 mm ID column P/N 206630.*Subject to quality of the sample and mobile phase conditions.
Long Term Reproducibility
Enduro C18 demonstrates excellent reproducibility* with minimal loss of peak shape and hence efficiency following 2,500 injections. Column life and reproducibility are all sample dependent it is always recommend that analytical column is protected with a guard column.
17Enduro GC and LC Columns
Enduro C18 Specifications
While Enduro is recommended as a “general purpose” reversed-phase column it is important to understand some of the fundamentals of liquid chromatography to extract the best performance from the Enduro column. The most important factors that influence the performance of a reversed-phase C18 LC column are:
• Bonding density – the amount of ligand per m² surface area typically between 3 and 4 µmol/m². It is often confused with carbon load which is a combination of phase and endcapping.
• Particle size – typically 5 and 3 micron spherical silica particles• Pore size and surface area – the typical range for C18 analytical columns is 6 to 30nm
(60-300 Å). The pore has to be big enough to comfortably accommodate the analyte molecule.
Stationary Phase (fully endcapped)
Particle Diameter µm
Pore Size Å Pore Volume m2 Specific Surface m2
% Carbon
Enduro C18 4.5 ± 0.8 135 ± 15 1.0 ± 0.1 300 ± 40 18.2
Enduro C18* and C8* LC Column Range
Dimension Column (120A pore size) PN
250 x 4.6mm C18 – 5 µm 206631
150 x 4.6mm C18 – 5 µm 206630
100 x 4.6mm C18 – 5 µm 206632
150 x 4.6mm C18 – 3um 206640
100 x 4.6mm C18 – 3um 206641
250 x 4.6mm C8 – 5 µm 206651
150 x 4.6mm C8 – 5 µm 206650
100 x 4.6mm C8 – 5um 206652
10 x 4.0mm Guard Column: C18 – 5 µm 250001
10 x 4.0mm Guard Column: C18 – 3um 250000
10 x 4.0mm Guard Column: C8 – 5 µm 250002
Enduro C8 5 μm phase and C18 3 μm phase will be available from July 1st 2011
Enduro GC and LC Columns18
The general purpose, robust nature, of the Enduro C18 means the column is well suited to Method Development. Here is a useful guide to help you develop your HPLC method on Enduro C18.
Gather Information
Study Chemistry of Substance
Published Method?
No
Is Substance Charged?
No
Start with Enduro C18 on partially degraded* product or
resolution mixture/standard
Adjust pH >1.5 Away from pKa Value
Yes
Run Gradient with 5-90% Acetonitrile
All Peaks Resolved?
No
Look for Critical Peak Pairs
Change Parameters: - Flow Rate - Temperature - pH - Gradient Profile to
Resolve Close Peaks
All Peaks Resolved?
No
Change Organic Modifier to MeOH or THF.
Consider Ion-pairing Reagents
All Peaks Resolved?
No
Change to Different RP Column, Phenyl, C8, CN, etc.
All Peaks Resolved?
No
Change to Different LC Mode Normal Phase, IEC, etc.
All Peaks Resolved?
No
LC is Not Suitable for the Analysis
Yes
Yes
Perform Run under Conditions of the Published Method
Yes
Yes
Yes
Yes
Large Gaps Between all Peaks?
No
Optimize Method for Short Analysis Time
Parts With No Peaks?
No
Check for Method Robustness and Critical
Parameters
Method Finished - Documentation
Method Validation
Yes Increase Slope of Gradient, Use Shorter Column
YesPartially Increase Gradient
Slope to Skip Areas With No Peak
Does the Method Work on the Real Sample?
Yes
No
*Alakaline, acid, UV, heat or peroxide degradation
Enduro C18 is Ideal for Method Development
19Enduro GC and LC Columns
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