medical laboratory technology

1. High performance
Liquid
Chromatography
(HPLC)

2. principle
• Sample to be separated is allowed to pass through a
stationary phase, due to high pressure applied, based on
the size sample to be separated will be comes out of the
stationary phase
Stationary phase; column
Mobile phase; solvent

3. DATA
RECORDER
filter

4. COMPONENTS
•Solvent system
•It is based on the sample to be separated
•Solvent system should be free from
contaminations

5. •Pumps
•Used to produce appropriate pressure to
push solvent and sample into the column

6. •Sample injector
Automatic, manual

7. •Filters
•To remove impurities in sample/solvent
•Column
separate sample into different components

8. •Detectors
•UV
•Fluroscence

9. Data recorder
Records the data

10. Working
•Solvent and sample is pushed to column by high
pressure produced from pumps
•Sample is applied through sample injection loop
•Sample to be separated is passed through the
stationary phase
•Sample is separated And comes out of the column
•Then detected by the detector
•Data recorder Records the data

11. Rt

12. •Retention time (Rt); time period from the
sample injection to the appearance of 1st
signal (peak)

13. applications
•Separation of aminoacids
•Separation of proteins
•Measurement of glycated Hb