Help with working a fluorescence detector!!!
Posted: Thu Apr 17, 2008 4:41 am
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!
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!