Indian Council of Agricultural Research
(Agricultural Knowledge Management Information System)


ICAR-Central Coastal Agricultural Research Institute, Ela, Old Goa, Distt. North Goa, Goa-403402 (https://ccari.icar.gov.in/)
Click Here to See Management Committee
Instrument Details
Instrument/Facility Name: HPLC (High performance liquid chromatography)
Make                                   : Agilent
Model                                  : 1200 Infinity Series
Specification   :Consists of Auto sampler, Diode Array Detector with column 16 and 18

Click to ViewHPLC (1200 Infinity Series of Agilent) is an high performance equipment for identification and quantification of different molecules in the samples. This modular design has wide range of solvent delivery, injection and detection options and suitable for configuration as per requirement of samples. It has capacity at purification at micro, analytical or prep scales and suitable for high throughput labs. Fitted with Diode Array Detectors achieves a peak to peak noise level with utmost accuracy.

  Working Principles
Click to ViewHigh performance liquid chromatography (HPLC) is basically a highly improved form of column liquid chromatography. Instead of a solvent being allowed to drip through a column under gravity, it is forced through under high pressures of up to 400 atmospheres. That makes it much faster. All chromatographic separations, including HPLC operate under the same basic principle; separation of a sample into its constituent parts because of the difference in the relative affinities of different molecules for the mobile phase and the stationary phase used in the separation.

  Applications
  User Instructions
The information that can be obtained by HPLC includes resolution, identification and quantification of a compound. It also aids in chemical separation and purification. The other applications of HPLC include : Food and Flavour, Pesticide residues, Pollutant levels etc.HPLC Operating Instructions: Prime the pumps by drawing the solvent (Pump B), Power on the HPLC. Invoke the program. Ensure loading device is set to ‘L’ (for load). Use a blunt needle syringe for injections. Wash syringe a few times with your sample, tap-out/remove any air bubbles in the syringe. Draw about 0.5mL of the ‘test’ sample into the syringe Inject the sample into the loop of the injector, please do not force the needle; leave the needle in the injector. Only a volume equal to that of the loop size will be injected on to the column at the running of the chromatogram. Make sure there is sufficient amount of the solvent in the solvent reservoir and the eluate ends up in the waste container. Running the Program: Once the separation is complete use the vertical scroll bar of the upper left window, section next to the graph, to zoom in/out on the peak Click ‘Post Run’ button, a cursor appears on the chromatogram window. Place the cursor on top of the peak(s). Left click to place a tag on the top of the peak(s) of the chromatogram. Watch the information on the top left window for the highest absorbance of the peak, before selecting the peak. At this time you may save the data file; File : Export Data. Click ‘Tags’ button; UV absorbance at the peak(s) of the eluted components selected in the preceding step appear in a table. This value of absorbance is proportional to the concentration of the analyte (plug) flowing through the detector.. To terminate the session; click ‘Stop’ in the program, Exit Program Turn off HPLC power.

  Contact Us
Contact No. 0832-2284678 Extension - 215
Email Id thangam.m@icar.gov.in

  HPLC (High performance liquid chromatography) Charges
Procedure Name
Industry
University
National LAB/R&D
Remark
Resolution, identification and quantification of solvent soluble compound
Rs. 5000 p
Rs. 1000 p

Additional Rs 100/-towards postal charges if result are to be posted
The Demand Draft should be in favour of "ICAR UNIT: CCARI, GOA Payable at goa"
Letter, DD & Samples send to "To, The Director, ICAR-Central Coastal Agricultural Research Institute, Ela, Old Goa - 403 402, Goa "