1. Chromatography Integration and Trouble-shooting
Presented by: Marlene Moore
October 2007
Advanced Systems, Inc.

2. Agenda
Problems
Tools
Solutions
Advanced Systems, Inc.

3. Problems Baseline Disturbances Irregular Peak Shapes, Sizes
Retention Time Shifts
Loss of Separation or Resolution
Quantitation Difficulties
Rapid Column Deterioration
Ghost Peaks
Broad Solvent Front
Advanced Systems, Inc.

4. Baseline Disturbances
spiking
noise
wander
drift
offset
Advanced Systems, Inc.

5. Spiking Particulate Matter Passing Through Detector Loose Connections
Cables
Circuit boards
Advanced Systems, Inc.

6. Noise Contaminated Injector/column Physical Defect Detector
Defective Detector Board
Carrier Gas Leak at Septum or Column
FID, NPD, FPD
Column in Flame
Incorrect Combustion Gases or Flow Rate/leak
ECD, TCD
Air Leak
Advanced Systems, Inc.

7. Wander Contaminated Carrier Gas Contaminated Gas Chromatograph
Isothermal Conditions
Contaminated Gas Chromatograph
Poor Control of Carrier Gas or Detector Gas Flows
Poor Thermal Control of Detector
Advanced Systems, Inc.

8. Drift Septum Bleed Upward Downward GC or column contamination
Damaged stationary phase
Downward
Incomplete conditioning of column
Unequilibrated detector
Advanced Systems, Inc.

9. Offset Contamination: Column Inserted Into Flame (FID, NPD, FPD)
Injector, column, detector
Carrier gas - change/install impurity traps
Column Inserted Into Flame (FID, NPD, FPD)
Malfunctioning or Improperly Set Recording Device
Advanced Systems, Inc.

10. Advanced Systems, Inc.

11. Peak Shape Peak Shape tells the story of your chromatography
Advanced Systems, Inc.

12. Irregular Peak Shapes tailing fronting split reduced rounded negative
Advanced Systems, Inc.

13. No Peaks Plugged Syringe Broken Column
Injecting Sample Into Wrong Injector
Column Installed in Wrong Detector
Integrator Reading Wrong Signal or Not Connected
Detector Not on
Temperature
Oven not heated
Column temperature too low
Injection port temperature too low
Detector Gases Improperly Set or Not on
Very Low or No Carrier Gas Flow
Advanced Systems, Inc.

14. Reduced Peaks Plugged Syringe Change in Injection Technique
Large Leak - Injector
Split Ratio Too High
Too Short of Purge Activation Time for Splitless Injections
Very High Septum Purge Flow
Too Low Injector Temperature
Column Temperature Too Low
Sample Components Absorbed by Column or Inlet Liner
Advanced Systems, Inc.

15. Reduced Peaks
Initial Temperature Column Too High for Splitless or On-column Injections
High Background Signal -
Contamination
Excessive column bleed
Check auto zero function and setting
Improperly Operated Detectors
Impurities Carrier Gas or Detector Gas
Detector Compound Mismatch
Excessive Attenuated Integrator Signal
Sample Concentration or Integrity Problems
Advanced Systems, Inc.

16. Tailing Peaks Active Injector Liner or Column Contaminated
Dead Volume Caused by Poorly Installed Liner, Column or Union
Poorly Cut Column End
Polarity Mismatch - Stationary Phase, Solute or Solvent
Cold Spot in the Flow Path
Advanced Systems, Inc.

17. Tailing Peaks Solid Debris in Liner or Column
Poor Injection Technique - Too Slow
Too Low of a Split Ratio
Overloading on a PLOT Column
Decrease sample amount
Amines and carboxylic acids tail on most columns
Need ph modified stationary phase
Some Peaks Always Exhibit Tailing
Advanced Systems, Inc.

18. Rounded or Flat Topped Peaks
Overloaded Detector
Exceeding Range of Integrator
Advanced Systems, Inc.

19. Split Peaks Detector Overloaded Poor Injection Technique
Poorly Installed Column Into Injector
Column Temperature Fluctuations
Coelution of Two or More Compounds
Mixed Sample Solvent for Splitless or On-column Injections.
Advanced Systems, Inc.

20. Negative Peaks All Peaks Negative - Polarity Check Connectors
TCD - Select Peaks
Thermal conductivity of compound greater than carrier gas
ECD - After Positive Peak
Dirty or old ECD cell
Advanced Systems, Inc.

21. Retention Time Shifts Different Column Temperature
Different Carrier Gas Flow or Linear Velocity
Leak in Injector - Septum
Contaminated Column
Change in Sample Solvent
Advanced Systems, Inc.

22. Loss of Separation/Resolution
Contaminated Column
Damaged Stationary Phase
Different Column Temperature, Carrier Flow Rate, Column
Large Changes in Sample Concentrations
Improper Injector Operation
Advanced Systems, Inc.

23. Quantitation Difficulties
Injection Techniques
Split Discrimination
Different Purge Activation Time for Splitless Injection
Baseline Disturbances
Improper Integrator Settings
Inconsistent Detector Gas Flows or Temperatures
Column or Liner Activity
Advanced Systems, Inc.

24. Rapid Column Deterioration
Exposure of Column to Air at Elevated Temperatures
Exceeding Upper Temperature Limit of Column
Chemical Damage
Contamination of Column With High Molecular Weight Materials
Column Breakage
Advanced Systems, Inc.

25. Ghost Peaks Contamination Septum Bleed Dirty Syringe Sample Too Large
Injector
Column
Septum Bleed
Dirty Syringe
Sample Too Large
Previous Run Terminated Too Soon
BOO!
Advanced Systems, Inc.

26. Broad Solvent Front Poorly Installed Column Leak in the Injector
Solvent Interacting with Detector
Too Low
Split Ratio (Flow)
Injector Temperature
Too Long Purge Activation for Splitless Injections
Large Injection Volume
Low Column Temperature and High Boiling Solvent
High Column Temperature and Low Boiling Solvent
Advanced Systems, Inc.

27. Advanced Systems, Inc.

28. Tools Flowmeter Syringe (NEW) Methane or Nonretained Compound
Septa, Ferrules, Injection Liners (NEW)
Leak Detector
Column Test Mixture (Reference)
Checkout Column
Instrument Manuals
Advanced Systems, Inc.

29. Solutions Quickest Solution ---- Improve Chromatography ---
Do Manual Integration ---
Get Results out Fast !!
NO!
WRONG ANSWER!!!
Advanced Systems, Inc.

30. Solutions Preventive Maintenance Allow Time for Optimizing System
Clean detector
Clean injector
Clean and repair parts
Clean and tighten connections
Follow instrument manual suggestions
Allow Time for Optimizing System
Integration parameters
Programs
Advanced Systems, Inc.

31. Does anyone have a question?
Where do you document your trouble shooting?
Advanced Systems, Inc.