What is HPLC method?

High-Performance Liquid Chromatography (HPLC) and Ultra Performance Liquid Chromatography (UPLC) are both liquid chromatography procedures used to separate and detect the different components found in complex mixtures.

What is full form of HPLC?

HPLC is an abbreviation for High Performance Liquid Chromatography. “Chromatography” is a technique for separation, “chromatogram” is the result of chromatography, and “chromatograph” is the instrument used to conduct chromatography.

How do you perform an HPLC analysis?

For setting up the HPLC machine:

  1. Make sure you have all your buffers set up.
  2. Open the purge valve and purge the system for 5 minutes.
  3. Add your samples into the autosampler tray.
  4. Stop the purge.
  5. Close the purge valve.
  6. Run the system at a normal flow rate (1 ml/min) with your buffer to equilibrate the column for 10 minutes.

What are the types of HPLC?

Types

  • Partition chromatography.
  • Normal–phase chromatography.
  • Displacement chromatography.
  • Reversed-phase chromatography (RPC)
  • Size-exclusion chromatography.
  • Ion-exchange chromatography.
  • Bioaffinity chromatography.
  • Aqueous normal-phase chromatography.

What is HPLC calibration?

Know the procedure to calibrate the High Performance Liquid Chromatography ( HPLC) including leakage test, flow rate, reproducibility and linearity, lamp energy and pump pressure drop in Pharmaceutical Quality Control.

Why is HPLC used for?

The purpose high performance liquid chromatography (HPLC) analysis of any drugs is to confirm the identity of a drug and provide quantitative results and also to monitor the progress of the therapy of a disease.

What is column in HPLC?

In HPLC chromatography, the column is where the separation of the sample takes place. This is where the sample is passed through the stationary phase with the mobile phase, resulting in the separation of the sample components.

How do you prepare samples for HPLC?

The simplest form of sample preparation is direct injection of the analyte into the HPLC system. Even if this is possible there will be sample preparations required unless the analyst can sample directly from the sam- ple container, which is rare.

How do you calculate concentration in HPLC?

concentration of sample= Area of sample/ Area of standard x concentration of standard .

What is normal phase column?

When chromatographic separation is done in a normal phase mode, the surface chemistry of the stationary phase has a polar characteristic. The mobile phase is generally nonpolar organic solvent (such as hexane or heptane).