HPLC..... High Performance Liquid Chromatography Parts of HPLC Trouble shooting in HPLC Detector

1. High Performance Liquid Chromatography
Presented By
Varsha Jindaniya

3. What is HPLC??
• HPLC is high Performance Liquid Chromatography
• It is an important analytical method used to seperate the mixture of
compounds.
• It is an qualitative and quantitative method.
Instrumentation of HPLC
Solvent Reservoir:- used to storing the mobile phase

4. •Pupm:- An HPLC pump is an solvent delivery system. The function
of the pump is to maintain a constant flow of mobile phase through
the HPLC system. Pump is to help in flow the mobile phase through
the column with the help of high pressure.
A pump forces a solvent With the help of the high pressure up to
400 atmosphere through a column.
There are two types of HPLC pumps: reciprocating pumps and syringe pumps.
Injector:- Injector is inject the sample from the sample vials .

5. •Column:- column is an work as a heart of HPLC .Column is responsible
for seperation of mixture.
• Different Types of HPLC Columns Used in Analysis
• Normal Phase Columns.
• Reverse Phase Columns.
• Ion Exchange Columns.
• Size Exclusion Columns.
•Detector :- In HPLC the detector
• Is used to detect the seperated
• Components of mixture of
• Compounds.

6. Trouble shooting is observed in HPLC
1. Sharp peak is not found.
2. Pressure fluctuation .
3. Sample preparation is not right.
4. Standard sample and test sample is not working properly.
5. Something is wrong in PH of mobile phase .
6. System is not working properly.
Note :- This graph is showing tailing in
Peak . This type of peak is not acceptable.
Note :- If tailing limit is less than 2 %
Then peak is acceptable (NMT 2%)

7. Theoretical Plates should be greater than
1500 (NLT 1500).
This type of peak is acceptable.This is sharp
peak and theoretical Plates showing also.
In this graph the peak is sharp and this type of peak is also
acceptable.

8. Resolution in HPLC :- It is ability to seperate the two peaks.
Resolution limit is less than 2%
This graph is showing the proper resolution
between the two peaks.
This graph is not show the proper resolution in two
peaks . Both peaks are mearge together. This type of
peak is not acceptable.

9. Thank you