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Problemas con detector de fluorescencia HPLC Agilent 1200

Discussions about HPLC, CE, TLC, SFC, and other "liquid phase" separation techniques.

5 posts Page 1 of 1
Hola, estuve haciendo una curva de calibración con varios analitos de estudio, también realice la validación y pruebas en muestras reales. Después vino el service a hacer el mantenimiento, y cuando retomo las pruebas que venia haciendo en las mismas condiciones que venia trabajando, los valores de las áreas de fluorescencia disminuyeron a la mitad comparado con antes del service. En teoría el service me dijo que lo que realizo no debería haber afectado los valores. Con el detector de absorbancia los valores se mantuvieron antes y despues del service y solo se altero en fluorescencia. Realice varias tareas para ir descartando posibles problemas, se mencionan abajo:

Procedí a limpiar la lampara.
Probé con dos concentraciones (una al principio y otra al final de la curva de calibración), de dos compuestos fluorescentes (uno eluyó al principio y otro al final).
Hice varias pruebas con mi sistema de estudio en las condiciones que siempre trabaje (antes y despues del service)
Aun no pude resolver de donde viene el problema del detector de fluorescencia, si alguien puede darme una mano para poder finalizar mi trabajo, se lo agradezco de antemano.


Saludos,


Soledad Moyano
Are you sure that all the method parameters, including the PMT gain, are the same as before?
yes, of course. I have looked all parameters besides I have saved the method in the computer of the apparatus and I havent modificated it....
If you want to find out the cause of the change in peak areas, you have to determine what the service engineer exactly did. Did he change the lamp in FLD?

Altered peak areas are not a problem if the system suitability check of your method is OK. You simply have to make a new calibration and continue working.
Hi Soledad,

The first thing I would do (if possible) would be to test the very same conditions in a different system with exact same setup - maybe a nearby lab/department has one? - this could help confirming whether it is an instrument or not

If this is not possible i would like to understand more about your situation:
- what is your setup - do you have both UV and FLD in the same instrument? what comes first?
- what is your mobile phase?
- what is your sample and how do you do sample preparation?
- what column are you using?
Luccas Name

YMC applications specialist (LATAM)
5 posts Page 1 of 1

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