Cromatografia Hplc

TROUBLESHOOTING IN HPLC : 
TROUBLESHOOTING IN HPLC BY SHEKHAR SHINDE

WHAT IS TROUBLE SHOOTING ? : 
WHAT IS TROUBLE SHOOTING ? Troubleshooting  is a form of problem solving most often applied to repair of failed products or processes. It is a logical, systematic search for the source of a problem so that it can be solved, and so the product or process can be made operational again.Troubleshooting is needed to develop and maintain complex systems where the symptoms of a problem can have many possible causes.

WHAT IS TROUBLE SHOOTING ? : 
WHAT IS TROUBLE SHOOTING ? Troubleshooting requires identification of the malfunction(s) or symptoms within a system. Then, experience is commonly used to generate possible causes of the symptoms. Determining which cause is most likely is oftena process of elimination - eliminating potential causes of a problem. Finally, troubleshooting requires confirmation that the solution restores the product or process to its working state.

WHAT IS TROUBLE SHOOTING ? : 
WHAT IS TROUBLE SHOOTING ? In general, troubleshooting is the identification of, or diagnosis of "trouble" in a [system] caused by a failure of some kind. The problem isinitially described as symptoms of malfunction, and troubleshooting is the process of determining the causes of these symptoms. This is the short but brief idea about the Trouble shooting.

TROUBLE SHOOTING ON HIGH PERFORMANCE LIQUID CHROMATOGRAPHY : 
TROUBLE SHOOTING ON HIGH PERFORMANCE LIQUID CHROMATOGRAPHY There are many causes in which problem are occured in HPLC. By Instrumental. By Procedure.By Human Errors. Because of this causes user have to face various problem such as Back pressure, Leakage, Change in Retention Time, peak shape and base line problem.

CONTENTS : 
CONTENTS START UP-PRELIMIARY CHECKS COLUMNS AND FITTINGS LEAKS CHANGE IN RETENTION TIME BASELINE PRESSURE PEAKS EQUILIBRATION SENSITIVITY

START UP PRELIMINARY CHECKS : 
START UP PRELIMINARY CHECKS PROBLEM:- NoPeaks or Very Small Peaks. POSSIBLE CAUSE:- Detector off. Broken Connections to the Recorder. No sample/Wrong sample. Wrong settings on recorder or detector.

START UP PRELIMINARY CHECKS : 
START UP PRELIMINARY CHECKS SOLUTION:- Check detectors. Check Connections. Check Sample. Be Sure It Is Not Deteriorated. Checks For Bubbles In The Vials. Check Attenuation, check Gains.

START UP PRELIMINARYCHECKS : 
START UP PRELIMINARY CHECKS PROBLEM:- No Flow. POSSIBLE CAUSE:- Pump Off. Flow Interrupted. Leak. Air Trapped in the System.

START UP PRELIMINARY CHECKS : 
START UP PRELIMINARY CHECKS SOLUTION:- Start Pump. Check reservoirs. Check position of the inlet tubing. Check loop for obstruction or air. Check degassing of mobile phase. Check compatibility of the mobile phase components.Check fittings. Check pump for leaks and precipitates. Check pump seals. Disconnect column and prime pump. Flush system with 100% methanol. Contact servicing if necessary.

COLUMN AND FITTINGS LEAKS : 
COLUMN AND FITTINGS LEAKS PROBLEM:- Column End Leaks. POSSIBLE CAUSE:- Loose Fitting. SOLUTION:- Tighten or Replace Fitting Cut Tubing and Replace Ferrule; Disassemble Fitting, Rinse andReassemble.

COLUMN AND FITTINGS LEAKS : 
COLUMN AND FITTINGS LEAKS PROBLEM:- Leak at Detector. POSSIBLE CAUSE:- Detector-seal Failure. SOLUTION:- Replace Detector Seal or Gaskets.

COLUMN AND FITTINGS LEAKS : 
COLUMN AND FITTINGS LEAKS PROBLEM:- Leak at Injection Valve. POSSIBLE CAUSE: Worn or Scratched Valve Rotor. SOLUTION:- Replace Valve Rotor.

COLUMN AND FITTINGS LEAKS : 
COLUMN ANDFITTINGS LEAKS PROBLEM:- Leak at Pump. POSSIBLE CAUSE:- Pump Seal Failure. SOLUTION:- Replace Pump Seal; Check Piston for Scratches And, If Necessary, Replace.

CHANGE IN RETENTION TIME : 
CHANGE IN RETENTION TIME PROBLEM:- Changing Retention Times. POSSIBLE CAUSE:- Contamination Buildup. Equilibration time insufficient for gradient run or changes in isocratic mobile phase.

Slide 16: 
First few...