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HPLC trouble shooting - Glutamate estimation (Apr/20/2009 )

Dear all
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



need more information.

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.


Relation between fever and outcome in injured victims of an internal armed conflict: the experience in a war surgery hospital of the International Committee of the Red Cross; Korver AJ; The records of 1,941 consecutive patients-victims of an internal armed conflict-admitted and treated over a period of 12 months at the surgical hospital of the International Committee of the Red Cross are analyzed retrospectively . They are divided in two groups: those with "fever" and those with "no fever." Patients are classified as having fever when the orally measured temperature is 39.0 degrees C or more for 2 or more subsequent days . The study indicates that fever is associated with the outcome in terms of an increase in the number of operations, the use of blood, the number of days in the hospital, and mortality, Surgeons working in internal armed conflicts without microbiological testing facilities have to consider fever as an important indicator for the clinical condition of their patients.