Which standard is used for HPLC calibration?
HPLC Calibration – GMP SOP This HPLC calibration Standard Operating Procedure (SOP) describes all the individual steps necessary for calibrating a HPLC in accordance with cGMP (current good manufacturing practices).
Why caffeine is used in HPLC calibration PDF?
Caffeine is convenient because it is not volatile, readily available, safe, has strong UV adsorption if you are using UV detector, and does not retain too long on RP columns (faster calibration).
How do you do gradient calibration in HPLC?
Calibration of the Gradient Proportioning Valve (GPV) Fill the reservoir A and B with 100% Methanol. Fill the reservoir for the C and D solution containing caffeine in Methanol. Prime the pump using the solvent from all the lines individual for about 5 min with flow rate of 3.0ml. Connect the union in place of column.
What are HPLC parameters?
The HPLC parameters are the: efficiency factor(N), the retention factor (kappa prime), and the separation factor (alpha). Together the factors are variables in a resolution equation, which describes how well two components’ peaks separated or overlapped each other.
What is calibration as per ICH guidelines?
Definition of Calibration: ICH The demonstration that a particular instrument or device produces results within specified limits by comparison with those produced by a reference or traceable standard over an appropriate range of measurements.
Why uracil is used in HPLC calibration?
Uracil is used for determination of the column void (dead) volume in reverse-phase chromatography, or in other words it is used to determine the dead time (the time for the unretained species to reach the detector).
Why is caffeine used in HPLC?
Caffeine is cheap, stable, safe and can be shipped without restrictions. It is not the best choice for wavelength calibration, but for injector precision and detector linearity it works well.
What is RT in HPLC?
Retention time (RT) is a measure of the time taken for a solute to pass through a chromatography column. It is calculated as the time from injection to detection.
What is calibration period?
A calibration interval is the period of time established for a test instrument to be returned for recalibration. We recalibrate instruments so we can be confident they are performing as good as, or better than, described in the manufacturer’s specifications.
What is the SOP for HPLC?
This SOP is applicable for the procedure to be followed in HPLC analysis and documentation and to ensure that Good Laboratory Practices are followed in the HPLC analysis.
How to operate and calibrate high performance liquid chromatography (HPLC)?
This SOP describes the procedure for operation and calibration of High Performance Liquid Chromatography (Model Shimadzu LC-2010 CHT). Ensure the calibration of HPLC before Use. Refer the manual for detail operation. Switch “ON” mains of instrument and PC. Switch “ON” instrument by switch provided on front side.
How do I calibrate my HPLC pump?
HPLC Calibration Procedure 1. Ensure that, the instrument is ready for calibration and Start-up procedure is followed. 2. Place inlet tubing of the Pump into the Water HPLC grade through a suction filter. 3. Allow the mobile phase to flow for about 5 min. 4. Block Pump outlet with the block screw.
What is the SOP for the calibration of the instrument?
Calibration, operation, cleaning, and precautions shall be done as per the respective instrument operation SOP Each individual module shall be calibrated as per the defined calibration frequency.