Chromatography Forum

I am using Waters 996 PDA Detector. I need to perform qualfication in the operating range of 210 nm to 270 nm. Please let me know if you know of any compounds and working concentration to evalute the PDA performance in this operating range. I tried Anthracene and Caffeine and did not have much luck. I was using about 100 mg anthracene in 500 mL acetonitrile and could not get any spectrum. If you have an exact procedure, please let me know.
Thanks

holmium oxide gives several sharp peaks at 240nm to 650nm

Dear USAllevyn,

You may want to consult the following reference;

B. Esquivel, "Wavelength Accuracy Testing of HPLC Detectors", Chromatographia, Vol. 26 (1988), pages 321-323

There is a description of the use of an Er solution that has several sharp bands in the region you mention. This solution will let you check wavelength accuracy and spectral resolution. Waters used to ship this solution with their Uv detectors. I do not know if they still do.

if you have any problems using the solution, let me know. I am very familiar with its use and practical details.

Good Luck,

josebenjamin

Benzophenone, Methyl paraben or propl paraben can also be used

I was using about 100 mg anthracene in 500 mL acetonitrile and could not get any spectrum.

Please let me know:
You use this solution as injected sample solution or as the component of mobile phase? If the first, it is very odd that you did get the spectra.
If the solution is a component of mobile phase, please don't use the method anymore. Use mobile phase without antracene and inject the sample that contains antracene.
Antracene has a sharp spektra with the peak in the wavelength of 251 nm+/- 2 nm.

Regards,
SYX

Try running caffeine without a column. Use about 5M of .01 PEEK tubing and 1ml minute H20. You should get lambda max at 205 and 273.