HPLC assay of 1,2,4,5-tetra-aminobenzene

3 posts / 0 new
Last post
bijaykp
bijaykp's picture
HPLC assay of 1,2,4,5-tetra-aminobenzene

 Iam planning to work on focal adhesion kinase (FAK) inhibitor, tetraaminobenzene, also called Y15. I couldn't find its HPLC assay method even after extensive literature search. If anybody else is working with it, please do suggest me the method.I will be very grateful. Thanks. 

Dr. Analytical
Dr. Analytical's picture
What HPLC equipment is

What HPLC equipment is available in your laboratory?  You will need an absorbance detector.  Investigate the best wavelength by measuring the absorbance spectrum.  I think you can use 220 - 240 nm.  Prepare a solution of Y15 at 100 ug/mL (0.1 mg/mL).

You should be able to separate this compounds with a C18 column and a mobile phase of acidified water and acetonitrile.  Prepare a solution of 0.1 % H3PO4 in HPLC grade water.  Mix the acidic water with acetonitrile in different ratios.  Start with about 90% acetonitrile and 10% acidic water and make an injection.  The peak should elute very quickly.  Increase the amount of acidic water until you get good retention.

If you do not get good retention, then you will have to try a more polar column.

Write to us with your results.

bijaykp
bijaykp's picture
 Thank you, Dr. Analytical.I

 Thank you, Dr. Analytical.I did a wavelenghth scan and the maximum absorbance was observed at 228-230 nm. I'm currently testing on different prpoprtion of acetonitrile and water as mobile phase. I'm really grateful. Thank you.