Trouble Shooting Guidance for Chromatography

S. No.	Problem	Probable Cause	Recommended Action
1	Noise	Injector contamination	Clean the injector or replace linear
		Detector Contamination	Clean the detector
		Contaminated or low quality gases	Use better grade gases, Check for expired gas traps or leakage.
		Incorrect combustion gases or flow rate	Adjust the flow rates to the recommended values or as per STP.
		Air leakage when using an MS, ECD or TCD	Find and eliminate the leakage.
		Old detector filament, lamp or electron multiplier	Replace appropriate port to resolve the issue.
		Septum degradation	Replace septum
2	Sequence Interrupted due to missing vial	Vail Not kept use of incorrect vial or Vial not detected or Vial alignment and carousel malfunction or Vial broken in Head space injection or loop system	Place the vial at proper position, append the sequence and re-inject all the solution from the bracketing standard before the interrupted sample, provided the system suitability and solution stability requirement are met.
3	System Suitability not within acceptance criteria	Instrument related Incidence	Rectify the error, calibrate wherever required and reload the sequence
3		Insufficient Solution in vial	Fill sufficient quantity in vial. Repeat the sequence and re-inject provided solution stability and system suitability requirement are met
4	Baseline drift	Change in carrier gas flow rate during the temperature program	Verify the carrier gas flow rate and temperature programming.
		Poor column installation	Reinstall the column
		Too low of a split ratio	Increase the split ratio
		Incomplete conditioning of column	Condition the column until a suitable baseline is achieved
		Un-equilibrated detector	Allow the detector sufficient time to equilibrate
		Septum bleed	Use higher temperature septum or analyse sample at lower injector temperature
		Insufficient column equilibration or condition	Equilibrate/condition the column and Repeat the sequence with same standard and sample preparation provided solution stability and system suitability requirement are met.
		Pressure Variation	Identify and rectify the probable cause, calibrate the instrument and repeat or append the sequence provided the solution stability and system suitability requirement are met.
		Air bubble in system or improper degassing of mobile phase	Remove the air bubble by adequate purging or degas mobile phase repeat or append the sequence provided solution stability and system suitability requirement are met
		Wrong grade of reagent used	Prepare fresh sequence and reload the sequence after use of proper reagents and solvent.
5	Loss of resolution	Decrease in separation, Decrease column temperature	Check column temperature
		Different in column dimension or phase	Verify column Dimensions.
		Increase in peak width., Change in carrier gas velocity	Check carrier gas velocity
		Change in the injector	Check the injector settings
		Change in the sample concentration or solvent	Verify the Sample preparation against STP.
6	Rounded or flat Topped peaks	Detector overload	Verify the sample preparation and concentration.
7	Split peaks	Poor column installation	Recut the column and re-install the injector
		Sample degradation in the injector	Verify the injector temperature and investigate.
		Detector overloaded	Verify the sample preparation and concentration.
		Column temperature fluctuation	Check the oven temperature
		Co-elution of two or more compounds	Check the sample preparation and operational procedure
8	Ghost and Extra Peaks	Dirty syringe	Clean the syringe properly and if required use new syringe.
		Septum bleed	Use higher temperature septum or analyze sample at lower injector temperature. Condition septum before use.
		Impurities from solvent	Use high quality grade solvents.
		Carry over from syringe rinse	Clean and rinse the syringe with solvent.
9	Carry Over Obtained	Wash option not incorporated or inappropriate wash solution used	Incorporate wash option or replace with appropriate wash solution and repeat the sequence provided solution stability and system suitability requirement are met.
		Wash option not incorporated or inappropriate wash solution used	Incorporate wash option or replace with appropriate wash solution and repeat the sequence. Provided solution stability are met.
		Buffer disposition at the needle seat	Clean the needle seat and repeat the sequence provided solution stability and system suitability requirement are met.
		Inadequate wash or rinse parameters	Modify the parameters. Repeat the sequence if solution stability and system suitability requirement are met.
		Insufficient volume in rinse vial	Ensure sufficient volume in the rinse vial and repeat the sequence provided solution stability and system suitability requirement are met.
10	Tailing peaks	pH of mobile phase not adjusted properly.	Verify the pH of mobile phase.
		Sample components absorbed by inlet liner or column	Replace with new deactivated liner or replace glass wool or packing.
		Co-elution of two or more compounds	Check the sample preparation and operational procedure
11	No peaks	Plugged syringe	Check the syringe or inject the sample with new syringe
		No carrier gas flow	Immediately lower the column oven temperature to 35°C. Verify the carrier gas flow rate. Check the leakage at column connection and septum. Pressure at gas source should exceed pressure needed at head of column by at least 15 psi at maximum temperature.
		Detector not ‘ON’ or gases improperly set	Check detector and gas settings
		Sample injected into the wrong injector	Re-inject the sample into proper injector
		Column installed into the wrong detector	Reinstall the column into the proper detector.
		Column broken	If broken at the inlet or the detector end, make clean cut and re-install column.
		Injector port temperature too low	Verify injector port temperature
		Oven not heated or column temperature too low	Verify column oven temperature
		Mobile phase port not selected properly or Drain valve open	Select proper port or close drain valve and repeat the sequence provided the provided solution stability and system suitability requirement are met
		Change in position of vial due to wrong vial labelling	Fill new vial with correct solution and append the sequence along with all the vials from the bracketing standard before the interrupted sample, provided solution stability and system suitability requirement are met
		Pressure drop due to air bubble trapping or cartridge problem	Purge the system with mobile phase to remove trapped air bubble or sonicate the cartridge. Saturate the column and re-inject the vials from the bracketing standard before the interrupted sample, provided solution stability and system suitability requirement are met
		Wrong injection type given in the software i.e. Blank is selected instead of standard or Unknown	Re-inject solution, provided solution stability and system suitability requirement are met.
		Leakage in system	Rectify the error, calibrate wherever required and reload the sequence
12	Abnormal peak area or Unusual Chromatogram	Leakage	Rectify the error, calibrate and reload the sequence
		Insufficient solution in vial	Fill sufficient quantity in vial. Re-inject provided solution stability and system suitability requirement are met
		Change in position of vial due to wrong vial labelling	Fill new vial with correct solution and append the sequence provided solution stability and system suitability requirement are met.
		Vial Filled completely till the neck	Re-inject from same solution.
13	System Communication error	Server problem or LAN connection error	Rectify the problem. Re-inject the vials from the bracketing standard before the interrupted sample provided the provided solution stability and system suitability requirement are met
14	Sequence interrupted due to power failure	Any module switched off or loose connection or faulty plug or UPS shutdown	Rectify the problem. Re-inject the vials from the bracketing standard before the interrupted sample provided the provided solution stability and system suitability requirement are met
15	Instrument hardware related error like “ Pump No Motor Home”, “ Buffer Overflow”, “Cooling fan Error” “Auto sampler Get hang”	Instrument hardware error	Rectify the problem. Re-inject the vials from the bracketing standard before the interrupted sample provided the provided solution stability and system suitability requirement are met