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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 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


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


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.



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.



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


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.



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.



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.



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


EN20 - bondable polyethylene glycol (Wax)

• Industry standard wax column.• Low bleed.• Polar phase suitable for hydrogen bonding analytes.



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.



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.



Part No.

0.10 0.10 10 10 to 330/350 053230

0.25 0.25 30 10 to 330/350 053231


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.



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.



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


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


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


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


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



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




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.


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


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.


0 1 2 3 4 5 6 7 8 9 10

Time (min)

2500th Injection (1185 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.


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


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

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Beijing Head OfficeRoom 917/918 Prime Tower,No. 22 Chao Wai Street, Chaoyang District, Beijing 100020Tel: 010-65888666Fax: 010-65886577

Shanghai OfficeRoom 513, Xiandai Jiaotong Building,No. 218 Heng Feng Road, Zhabei District, Shanghai 200070Tel: 021-51611815Fax: 021-51611816

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enduro gc and hplc columns - jjindustries.com.cn€¦ · enduro gc and hplc columns sge has five...

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