Chromatography Forum

Dear All,

I have some issue with my HPLC 1200 Injector program. Accordingly that I have not so match experience with Injector program but I need to set up it because I have to mix two component and inject they at Time 0 and the same vial after 60 minutes.

I followed the Agilent Instruction and I setted COMMAND like follow:

METHOD 1
-DRAW MAX AMOUNT (100uL) from vial 4 max speed.
-EJECT MAX AMOUNT INTO VIAL 12, MAX speed
-DRAW MAX AMOUNT FROM VIAL 2, MAX speed
-EJECT MAX AMOUNT INTO VIAL 12
-DRAW DEF AMOUNT FROM VIAL 12 MAX SPEED
-INJECT

Def amount was 1uL

METHOD 2 after 60 minutes
-DRAW DEF AMOUNT FROM VIAL 12 MAX SPEED
-INJECT

For time 0 I used METHOD 1 (total time for the course was 60 minutes).
After first course, 60 minutes happens, and in the sequence table I setted METHOD 2 on same vial.

Issue was that at Time 0 I had a very big peak (over 2000UA) and after 60 minutes I had nothing......

I put also a wash step after mixing and before inject in the method 1 but nothing.

Sorry for the long explanation but I'm in loop and I cannot go out.....

Thanks in advance

Antonio

Hello

It should work with programed injection...However, if you have defined injection volume for method 2 why do you use program for injection? Just set up standard injection with exact injection volume.
Perhaps you should check some other compounds (even simple LC checkout sample) just to confirm that sampler takes and injects sample with both programmed (injections) methods.

Regards

Tomasz Kubowicz