GC Column Care Guide

GC Column Installation............................................................................................. 1-2

GC Column Conditioning and Testing............................................................................ 3
GC Hints and Tips
Protecting the GC Column and Extending Life Time................................................... 4
Carrier Gas Selection and Flow Optimization............................................................. 5
Column Storage..................................................................................................... 5
Chemical Compatibility / Solvent Rinsing of Zebron™ Capillary Columns...................... 6
 GC Column Installation
A. Capillary GC Installation with a Traditional GC Nut
Note: GC columns do not have a specific directional flow when received from the manufacturer. Upon initial use of your new Zebron™ column, Phenomenex
recommends the practice of dedicating one specific end of the column for injector installation only. This is particularly important when dealing with active/caustic
or contaminating compounds. If these compounds are routinely injected onto the column, degradation of the phase will occur - leading to higher bleed. A typical
first step to remedying (removing) this bleed would be to trim 10 cm from the front (injector) end of the column and keep trimming this inlet end of the column as
necessary. Trying to remedy any bleed issues by trimming the column may not work if both ends have been interchangeably installed into the inlet.

Injector Installation Detector Installation

1. Place a capillary nut and ferrule on the injector end of the GC Note: For users with sensitive detectors such as MS and ECD, column
column, allowing a section of column to protrude. Trim one to conditioning steps should be performed before installing the column to
two centimeters (cm) from the protruding end to remove ferrule prevent contamination and frequent maintenance of the detector.
contamination that may have entered the column (Figure 1). Inspect 1. Place the column nut and ferrule past the end of the column and cut
the cut with a magnifier to ensure that a smooth, clean, square-cut a centimeter or two off the end of the column (Figure 1). Be sure
edge has been made – cut again if necessary (Figure 2). that the ferrule is the right size and pointing in the correct direction.
2. Carefully hang the column in the GC oven, being cautious not to Inspect the cut with a magnifier and ensure that the cut is square
scratch or damage the polyimide coating on the capillary tubing. and smooth (Figure 2). Cut again if needed.
Rotate the column to avoid sharp bends of the capillary column and 2. Insert the outlet end of the column into the detector exactly the
any contact of the column with oven surfaces. distance prescribed in the instrument manual. Distances will vary
3. Insert the column into the injector exactly the correct distance between detectors. Tighten the ferrule nut finger-tight then ½ turn
specified in the instrument manual. Tighten the ferrule nut finger- with a wrench. If the column can still be moved, tighten another ¼
tight then ½ turn with a wrench. If the column can still be moved, turn until the column is secure.
tighten another ¼ turn until the column is secure. 3. Inspect the column connections for leaks using an electronic leak
4. Adjust the carrier gas. detector. Leaks at the inlet end may introduce oxygen to the column
that will result in increased column bleed and damage to the column
phase.

Figure 1: Cutting Fused Silica Tubing Figure 2: Proper and Improper Cut Capillary End

score capillary with

smooth edge of
wafer at a 45° angle a b
© 2010 Phenomenex, Inc. All rights reserved.

4 Correct X Incorrect
apply force in a
downward direction
Examples of: (a) A Properly Cut Fused Silica Column End
(b) An Improperly Cut End
tubing should break
cleanly

inspect cut with a

magnifying glass

1 www.phenomenex.com
GC Column Installation
B. Capillary GC Installation with the Cool-Lock™ Nut
• Avoid burning your fingers again – cools with the oven ~exclusive~
• N o Wite-Out or Tipp-Ex needed
® ®

• A chieve the proper installation depth each and every time

• N o need for wrench with hand-tightened connections

patent pending

1 2 3

1. Insert column through column nut and 2. Hand-tighten the column nut into the gauge 3. Unscrew the installation gauge and install
ferrule. Cleanly cut ≥ 1 cm of the column until the ferrule starts to compress. Adjust the the nut and column into the instrument.
that is protruding from the nut and ferrule. length of the column to the desired length.
Insert the end of the column and column Further tighten the nut into the gauge until the 4. Make sure Cool-Lock Nut is in the full
nut into the bottom of the Cool-Lock Nut column cannot slide. If using the Cool-Lock extension position to achieve the correct
Installation Gauge. Nut, tighten its bottom ferrule. Make sure lower premeasured position, and allow for proper
body/ferrule is in the full extension position. leak checking.
Otherwise, mark the column at the base of the
column nut to ensure the correct installation
depth after installation.

Agilent® GC Systems Shimadzu® GC Systems

Cool-Lock GC Capillary Nut * Cool-Lock GC Capillary Nut **
Part No. Description Unit Part No. Description Unit
AG0-8319 Cool-Lock GC Capillary Nut For Use with ea AG0-8419 Cool-Lock GC Capillary Nut For Use with ea
Short-Style Ferrules Short-Style Ferrules
AG0-8320 Cool-Lock GC Capillary Nut For Use with ea AG0-8420 Cool-Lock Nut Installation Gauge ea
Long-Style Ferrules
**Guaranteed fit for Shimadzu 2010 and 2014 GC systems
AG0-8349 Cool-Lock Nut Installation Gauge ea
Replacement Ferrules
AG0-4701 GC Capillary Ferrules Graphite 1⁄16 in. to 0.5 mm ID 10/pk
AG0-4704 GC Capillary Ferrules Graphite 1⁄16 in. to 0.8 mm ID 10/pk

*Guaranteed fit for Agilent 5850, 5890, 6890, 6850 GC systems

www.phenomenex.com 2
 GC Column Conditioning and Testing
A. Column Conditioning Zebron’s Text Mix
1. Allow sufficient time for the carrier gas to flow through the column to • Suitable for Phenomenex Zebron™ and equivalent brands
purge any oxygen that may be in the system.
• C onvenient way to check column performance
2. Raise the temperature of the column to the maximum isothermal
• Affordable and easy-to-use
operating temperature that is listed on the individual Zebron™ GC
Column Test Report. Maintain this temperature until a constant baseline
is achieved. Conditioning times will depend on the phase identity and Zebron Phase Test Mix
thickness, with thicker films taking longer to stabilize. If necessary Part Number
in order to minimize the downtime of the instrument, columns can be ZB-1 AG0-5155
conditioned overnight at the maximum isothermal temperature.
ZB-1ms AG0-7805
B. Installation Testing ZB-1HT AG0-5155

Inject a detectable unretained sample, such as methane for an FID, to determine ZB-5 AG0-5155
dead volume time and linear gas velocity at the desired column temperature. ZB-5ms AG0-7578
Adjust gas pressure for optimal flow depending on carrier gas selection. ZB-5MSi AG0-8362
ZB-5HT AG0-5155
1. The non-retained peak must have ideal peak shape or installation is faulty
and needs to be redone. ZB-35 AG0-5156
ZB-35HT AG0-5156
C. Checking for Leaks ZB-1701 AG0-5156
Use a thermoconductivity detector to check for leaks. It is highly sensitive to ZB-1710P AG0-5156
H2, He, and N2 and will not contaminate the instrument or column. Liquid leak ZB-50 AG0-5157
indicators are not recommended for capillary columns. There is the risk of ZB-624 AG0-5159
drawing the liquid into the column or fittings and contaminating the system.
ZB-WAX AG0-5158
NOTE: If Vespel ferrules are being used, leakage can occur after the initial
® ZB-WAXplus™ AG0-7869
heating phase due to ferrule deformation. Be sure that the fitting is re-tightened ZB-FFAP AG0-5158
after this initial heating phase, and then carefully check all corrections for leaks.
ZB-XLB AG0-7578

D. Test Column with Zebron’s Text Mix ZB-XLB-HT AG0-7578

ZB-Drug-1 AG0-8431
Good chromatographic practices include the use of Performance Check
Standards to establish the baseline performance characteristics of your column Grob Test AG0-5154
and to monitor any changes over time. Test solutions should evaluate column
inertness, efficiency and resolution in an easy-to-use format. Most Phenomenex
test mixes are supplied ready to use at a concentration of 250 μg/mL in glass
ampules.

Match your column phase to the correct standard to set up initial running
conditions. Should you have any questions on the proper use of these Check
Standards, contact your local Phenomenex GC Specialists.

3 www.phenomenex.com
GC Hints and Tips
A. Four Key Steps to Protecting the GC Column and
Extending Lifetime
Figure 3. Some liners that are available pre-packed with glass wool or provide
It is important to protect the column and instrument components from additional column protection:
exposure to dirty samples. Non-volatile or high molecular weight Description Dimensions Material* Quartz Wool
GC Model No. ID x L x OD (deactivated) (Y / N) Mfr. No. Part No. Unit
components can contaminate the stationary phase, causing peak (mm)
resolution, lower accuracy, and poor column lifetime. Cutting off the
damaged portion will usually restore the column performance, but over Split/Splitless
time performance will degrade to a point where the column can no longer 5880/5890/6890 4 x 78.5 x 6.3 B (y) Y 210-4004 AG0-7515 5/pk
be used. If you are experiencing rapid degradation of column performance, AG0-7582 25/pk
there are several simple ways to help protect your column and increase
lifetime: Split/Splitless, Recessed Gooseneck Liner
5880/5890/6890 4 x 78.5 x 6.45 B (y) Y 5183-4691 AG0-4661 5/pk
1. Ensure Proper Sample Preparation 5183-4692 AG0-4662 25/pk

a. Filter your samples prior to injection with Phenex™ syringe filters (see
Appendix A for details). Cup Splitter/Split Liner
5880/5890/6890 4 x 78.5 x 6.3 B (n) N 5183-4699 AG0-4647 5/pk
b. Use Strata® or Strata™-X SPE Cartridges to eliminate contaminants 5183-4700 AG0-4648 25/pk
(see Appendix D for details).
Cup Splitter/Split Liner
2. Use a Guardian™ or Z-Guard AutoSystem™ 3.5 x 100 x 5 B (n) N 0330-5181 AG0-4663 5/pk
a. Guard Columns – Standard Guards. Z-Guard columns are 5 or 10
meter pieces of deactivated tubing that are connected to an analytical Splitless Single Taper/Liner
column using a glass press-fit connector. The tubing acts like a trap 5880/5890/6890 4 x 78.5 x 6.45 B (y) Y 5183-4693 AG0-4657 5/pk
for non-volatile residues that would otherwise damage the stationary 5183-4694 AG0-4658 25/pk
phase of your analytical column. * B = Borosilicate; Deactivated = Yes (y) or No (n)
b. Guardian Integrated Guard Columns. Unlike traditional guard
columns, there is no mechanical connection between the guard
and the analytical column. The result: all the benefits of a guard
column without the possibility of leaks or activity resulting from a
faulty connection. Please contact a Phenomenex GC Specialist for 4. “Baking Out” the Column
information on the Guardian. The easiest way to reduce column contamination is to add a short, high
temperature bake out at the end of the standard GC method. This bake out
3. Use a Wool Liner helps remove high boiling contaminants that would otherwise remain in the
The liner is the first line of defense for the column and the style column and cause damage. To bake out, the final oven temperature needs
chosen can make a big difference in how much contamination to be set high enough to ensure elution of these compounds, but not so high
gets onto the column. The easiest thing to do is to add a small as to cause thermal damage. This can be done either isothermally, or more
amount of silanized glass wool to a liner, which traps the non- commonly, via a gradient or ballistic increase until the last components
volatile compounds and prevents them from entering the column elute from the column.
(Figure 3). See Appendix B for other recommended GC accessories.
Caution: NEVER exceed the upper temperature limits of the column. DO
NOT exceed more than 15 minutes at the upper isothermal temperature
Caution: glass wool can also add activity for acids, bases, and limit specified for the column.
pesticides. Crushing the glass wool can lead to increased activity, so it is
recommended to purchase pre-packed liners, rather than try to pack your
own.

www.phenomenex.com 4
 GC Hints and Tips
B. Carrier Gas Selection and Flow Optimization
It is advisable to use the highest purity gas possible. Ultra high purity Optimal velocities uopt Optimal velocities uopt
(99.99 %), ultra pure carrier (99.995 %), or even research grade (99.9999 for low df values: (cm/sec) for high df values: (cm/sec)
%) is preferred to minimize critical impurities, instrument downtime and
H2 40 H2 25
troubleshooting. Three types of gas are commonly used as a carrier gas:
He 25 He 15
1. Hydrogen (H2): Hydrogen will yield maximal number of theoretical
plates for thin film columns and the high efficiency is largely retained N2 10 N2 7
at velocities higher than uopt. Hydrogen is not generally recommended
due to its hazardous nature.
2. Helium (He): When hydrogen is not used, helium is the best C. Column Storage
alternative for speed and sensitivity.
Important! The column may be left in the instrument for short-
3. Nitrogen (N2): Nitrogen is the last choice for thin film columns. For term storage. Ensure a flow of carrier gas through the column at
thick film columns, nitrogen yields the highest number of theoretical 100-200 °C. For long-term storage, disconnect from the GC and cap or
plates. However, the optimal velocity is fairly low (long analysis seal the capillary ends. Oxygen and moisture can degrade or irreversibly
times), and the loss in efficiency at higher velocities is high. If
damage the column, especially cyanopropyl-based phases. Wax
resolution is sufficient, hydrogen or helium are good alternatives.
(polyethylene glycol) and cyanopropyl-based phases are also susceptible
to UV-induced degradation and should be shielded from light (fluorescent
Figure 4: Carrier Gas Selection and Velocity Optimization Plots
or sunlight). Store the column in the original box. Upon reinstallation, cut
column ends to ensure that septum fragments or other debris have not
Carrier Gas Length
been left in the column.

H(u) curves for different gases H(u) curves for different

column lengths

Film Thickness Retention Factor

H(u) curves for different film H(u) curves for different k values
thicknesses

5 www.phenomenex.com
GC Hints and Tips
D. Chemical Compatibility / Solvent Rinsing of Zebron™ Capillary Columns
Important! Water and organic solvents such as those listed in Table 1 will not damage Zebron column stationary phases. However, inorganic acids and
bases should be completely avoided or rapid degradation and permanent damage to the stationary phase will result. In the event chemical damage is
incurred, the removal of 0.5-2 meter of capillary off the front end will often restore column performance. Select the rinse solvent from the Table below.

Table 1. Phase Compatibility with Rinsing Solvents

Phase Water Methanol CH2Cl2 CHCl3 Acetone Hexane

ZB-1, 1ms, 1HT

ZB-5, 5ms, 5MSi,

5HT

ZB-XLB,
ZB-XLB-HT

ZB-624

ZB-35, ZB-35HT

OK
ZB-1701,
ZB-1701P

ZB-50

ZB-WAXplus™ AVOID

ZB-WAX AVOID

ZB-FFAP AVOID

MR-1, MR-2

ZB-Drug-1

ZB-BAC1,
ZB-BAC2

ZB-Bioethanol

Table 2. Rinsing Conditions

Column ID (mm) Rinse Solvent Volume (mL) Pressure (psig)

0.25 5 40

0.32 5 40

0.53 10 20

Important: Rinse from the back to the front of the column to avoid pushing inlet contaminants further into the column.

www.phenomenex.com 6
 Appendix A
Phenex™ Syringe Filters

For Sample Filtration Prior to GC Analysis

• Increased column lifetime
• M ore consistent, reproducible results
• L ess system downtime

Syringe Filter Applications and Recommended Membranes

Application / Sample Recommended First Alternative Second Alternative
Fitler
HPLC and GC Sample Prep RC PTFE PES
Aggressive or Pure Organic Solvents PTFE RC NY
High Particulate Loads RC NY PTFE Try a Sample Pack!
Environmental Methods RC PTFE NY visit: www.phenomenex.com/sample
Food and Beverage RC PTFE NY
Clinical / Toxicology RC PES NY

Choose a pore size based on the nature of your sample and chromatographic method
Sample Description Recommended Filter Pore Size
General clarification of GC samples. 0.45 µm
Removal of fine pariculate matter prior to GC analysis. 0.20 µm

4 mm Diameter 15 mm Diameter 25-28 mm Diameter

for ≤ 2 mL sample volumes for 2 - 10 mL sample volumes for 10 - 100 mL sample volumes
Membrane Type/Size Part No. Unit Part No. Unit Part No. Unit
0.45 µm
Phenex-RC AF0-3103-12 100/Pk AF0-2103-12 100/Pk AF0-8103-12 1 100/Pk
(Regenerated Cellulose) AF0-3103-52 500/Pk AF0-2103-52 500/Pk AF0-8103-52 1 500/Pk

Phenex-PTFE 2 AF0-3102-12 100/Pk AF0-2102-12 100/Pk AF0-1102-12 100/Pk

(Polytetrafluoroethylene) AF0-3102-52 500/Pk AF0-2102-52 500/Pk AF0-1102-52 500/Pk

0.20 µm
Phenex-RC AF0-3203-12 100/Pk AF0-2203-12 100/Pk AF0-8203-12 1 100/Pk
(Regenerated Cellulose) AF0-3203-52 500/Pk AF0-2203-52 500/Pk AF0-8203-52 1 500/Pk

Phenex-PTFE 2 AF0-3202-12 100/Pk AF0-2202-12 100/Pk AF0-1202-12 100/Pk

(Polytetrafluoroethylene) AF0-3202-52 500/Pk AF0-2202-52 500/Pk AF0-1202-52 500/Pk

Please inquire with Phenomenex about other Phenex Syringe filters offerings.

Above syringe filters are non-sterile. Housing is made of medical-grade polypropylene (PP) and offer luer lock inlet/slip outlet connections, unless otherwise indicated.

1. 26 mm diameter.

2. Hydrophobic membrane. Can be made hydrophilic by pre-wetting with IPA.

7 www.phenomenex.com
Appendix B
Other Recommended GC Accessories

Gold Inlet Base Seals

• Prevents analyte adsorption
• Improves detection sensitivity
• Improves method reproducibility
• Fits all Agilent 5890/6890 split/splitless injection ports

Part No. Description Similar to Mfr. No.* Unit

Standard, single groove for splitless applications, 0.8 mm dia. inlet hole
AG0-7518 Gold Inlet Base Seal, splitless (single groove), 0.8 mm 18740-20885 2/pk
AG0-7519 Gold Inlet Base Seal, splitless (single groove), 0.8 mm 18740-20885 10/pk
High Split Flow, cross groove for split applications, 0.8 mm dia. inlet hole
AG0-7520 Gold Inlet Base Seal, split (double groove/cross), 0.8 mm 5182-9652 2/pk
AG0-7521 Gold Inlet Base Seal, split (double groove/cross), 0.8 mm 5182-9652 10/pk
Replacement Inlet Seal Washers
AG0-8397 Inlet Seal Washers, Gold Plated, for Agilent GC injection port — 12/pk

GuideRight™ Injection Hole Septa Long Graphite Ferrules

The GuideRight™ through-hole guides the needle during injection. Septum • High-purity graphite significantly reduces ferrule bleed
performance and lifetime increase while downtime due to bent needles or dirty • Special construction minimizes “flaking”
injection ports decreases. Choose from either high temperature PhenoGreen™ or • Stable to 450 °C - excellent for high temperature use
PhenoRed™, both rated to 400 °C.
Part No. Description Unit Part No. Description Unit
PhenoGreen 3⁄8 in. (9.5 mm) Diameter 0.4 mm Ferrule ID
AG0-7874 PhenoGreen 400 Injector Septa 50/pk AG0-4698 Graphite Ferrule 1⁄16 in. to 0.4 mm 10/pk
PhenoGreen 7⁄16 in. (11 mm) Diameter AG0-4699 Graphite Ferrule 1⁄16 in. to 0.4 mm 50/pk
AG0-7875 PhenoGreen 400 Injector Septa 50/pk 0.5 mm Ferrule ID
PhenoRed 3⁄8 in. (9.5 mm) Diameter AG0-4701 Graphite Ferrule 1⁄16 in. to 0.5 mm 10/pk
AG0-7916 PhenoRed 400 Injector Septa 50/pk AG0-4702 Graphite Ferrule 1⁄16 in. to 0.5 mm 50/pk

PhenoRed 7⁄16 in. (11 mm) Diameter 0.8 mm Ferrule ID

AG0-7917 PhenoRed 400 Injector Septa 50/pk AG0-4704 Graphite Ferrule 1⁄16 in. to 0.8 mm 10/pk
AG0-4705 Graphite Ferrule 1⁄16 in. to 0.8 mm 50/pk

Note: Not for use with GC/MS transfer lines.

www.phenomenex.com 8
 Appendix C
Vials

Headspace Vials
Round Bottom Flat Bottom
20 mL 20 mL
Compatible With The Following Autosamplers Headspace Autosampler Vials
Brand Model Round Flat Part No. Description 100/pk
Bottom Bottom AH0-7546 Headspace Vial 20 mL clear, round bottom,
Agilent HS 7694 • 23 x 75 mm
Carlo-Erba HS 850 • AH0-7547 Headspace Vial 20 mL clear, flat bottom,
Dani HS 39.50 / 86.50 • 23 x 75 mm
Fisons HS 850 • AH0-7548 Aluminum Seal, with PTFE/Silicone septum,
PerkinElmer HS 100 / 101 / 40 • 0.130” thickness
Tekmar-Dohrmann 7000HT • AH0-7550 Aluminum Seal, with Pharma-Fix molded
ThermoQuest HS 850, Trace GC • pressure-release septum
2000 + HS 2000 AG0-8350 Aluminum Seal, Magnetic Crimp Cap, 20 mm
Varian Genesis • • TEF/SIL, blue

Limited Volume Vials

QsertVial™ CD™ Vial

QsertVial kits, Clear wide-mouth vial, 12 x 32 mm, 300 µL volume, with CD Vial kits, Clear wide-mouth vial, 12 x 32 mm, 1.5 mL volume
write-on patch.
Part No. Description 100/pk Part No. Description 100/pk
AH0-7508 QsertVial Kit, Silicone/PTFE septa AH0-7509 CD Vial Kit, Silicone/PTFE septa
AH0-7505 QsertVial Kit, Silicone/PTFE septa with pre-slit AH0-7506 CD Vial Kit, Silicone/PTFE septa with pre-slit

9 www.phenomenex.com
Kit packs include caps.
Appendix D
Strata® and Strata™-X SPE Products
SPE products can help to eliminate contaminants and matrix effects,
resulting in increased signal-to-noise ratios and longer column lifetimes.
Phenomenex manufactures two types of SPE sorbents:

Product Offerings:
1. Strata - Silica Solid Phase Extraction (SPE) Products 2. Strata-X-Polymeric Solid Phase Extraction (SPE) Sorbents.
(Same functions as Strata but with the additional benefits):
• Quickly and easily cleans samples, concentrates analytes, and
facilitates solvent switching • De-conditioning resistant surface resists dry out and de-activation
and provides worry-free manual and automated processing
• Removes contaminants and eliminates matrix effects for improved
separation and column lifetime • pH stability from 1-14 for flexible method development
• A bility to load up to 66 % more sample per gram in comparison to
silica-based sorbents per gram
Formats: Strata and Strata-X SPE products are available in multiple formats:

Standard Tubes for Teflon®-Coated Tubes for Large Volume/Flash Tab-less Tubes
Traditional SPE Extra Inert Analysis Analysis Cleanup

96-Well Plates for High-Throughput On-line Extraction Cartridges Large Volume

20 x 2.0 mm

Need help selecting an SPE Sorbent? Please call your Phenomenex Sample Preparation Specialist for help or visit www.phenomenex.com/info/spehelp

Terms and Conditions

Subject to Phenomenex terms and conditions, which may be viewed at www.phenomenex.com/TermsAndConditions.
Trademarks
Zebron, Inferno, Cool-Lock, Phenex, Strata-X, Guardian, GuideRight, PhenoRed, PhenoGreen, and WAXplus are trademarks of Phenomenex, Inc. Strata is a registered trade-
mark of Phenomenex, Inc. Tipp-Ex is a registered trademark of Tipp-Ex. Agilent is a registered trademark of Agilent Technologies, Inc. Shimadzu is a registered trademark of
Shimadzu Corp. Wite-Out is a registered trademark of BIC Corporation. Vespel and Teflon are registered trademarks of E.I. du Pont de Nemours and Co. CD Vial and Qsert Vial
are trademarks of AR Corporation.
Strata-X is patented by Phenomenex.

© 2010 Phenomenex, Inc. All rights reserved.

www.phenomenex.com 10
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