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Help with working a fluorescence detector!!!

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

2 posts Page 1 of 1
I am currently attempting to setup an old fluoresence HPLC setup which has recently had a software upgrade to Class VP (latest edition). I have NO experience with fluorescence detectors of any type. This one is a Shimadzu RF 535 and I have used a reference from the literature for the method (for detection of doxorubicin - I have it in a water matrix as the ref. didn't say what to dissolve it in - using a 65% phosphate buffer/35% ACN mobile phase (1mL/min) with excitation at 480nm and emission 560nm).

I have run a baseline using MP but am getting nothing ie no noise at all and no peaks when a sample was run. It looks like there is no signal but I'm not sure why this would be. The detector voltage was at 2V and I have tried it at 3.333 but still no luck.

As I say I have no experience with fluorescence (and unfortunately no-one to teach me : ( ) so any help would be very much appreciated!

Thanks.

Merrin

P.S. if anyone could point me to a good online reference dealing with fluorescence chromatography that would be great too!

There is an old saying. 'If all else fails read the manual'!!

I would add and speak to the makers - Shimadzu.
No Tswett
2 posts Page 1 of 1

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