HPLC trouble shooting
Posted 20 April 2009 - 09:43 PM
I am trying to estimate glutamate via HPLC ECD methods. For the same i am using 0.1Mphosphate buffer ph 6 with 15% methanol as mobile phase and glutamate is derived with OPA (orthophthaldehyde) reagent composition [ 27 mg OPA + 1ml methanol + 12 ul mercaptopropanoic acid + 9ml of 0.1Mborate buffer ph 9.2]
now problem is that we are getting a very negative baseline which start after 1 min of injection and at the same time if glutamate peak is comming we are getting changes in RT after every injection
kindly suggest me about this problem or if any body is working on glutamate estimation by electrochemical detection methods plz provide me working protocol
Posted 21 April 2009 - 06:39 AM
injection size, column phase, equilibration volume, isocratic or gradient separation, etc.
you may be seeing the borate from the injection.
you may also want to post your question on the chromatography forum.
Edited by mdfenko, 21 April 2009 - 06:42 AM.
genius does what it must
i do what i get paid to do
Posted 30 April 2009 - 10:20 PM