1.0 PROCEDURE
1.1 CALIBRATION OF PUMP :
Start the HPLC pump as per IOP and check the flow rate by collecting water in a 10ml measuring cylinder. Using a stop watch set the time for 5 minutes and measure the volume collected for 1.0ml/ minute 1.5/minute and 2.0ml/minute flow rates respectively. Note the readings in duplicates.
1.1 CALIBRATION OF PUMP :
Start the HPLC pump as per IOP and check the flow rate by collecting water in a 10ml measuring cylinder. Using a stop watch set the time for 5 minutes and measure the volume collected for 1.0ml/ minute 1.5/minute and 2.0ml/minute flow rates respectively. Note the readings in duplicates.
1.2 CALIBRATION OF DETECTOR
To a clean, dry 100ml volumetric flask, add 0.1ml of Benzene and 0.1ml of Toluene. Mix by adding mobile phase and make up to volume. Inject equal quantities three times and record the ratio of the peak area.
Chromatographic conditions
Detector : 254nm
Flow Rate : 1.0 ml/minute
Mobile Phase : Methanol : Water = 80 : 20
Absorbance Scale : 0.02
Attenuation : 5
Injection Volume : 20micro litre
Column : C-18 (Shim-pak)
Area of first peak
Calculation : = Ratio
Area of second peak
2.0 DOCUMENTATION
2.1 F:Sxxxx/HCR - HPLC Calibration Record.
3 comments:
very useful and currect information u provided
this is useful matter to me.
thank you,
yours,
hajarathreddy.g.
what should be the ratio of benzene to toluene?
Post a Comment