Chromatography Forum

Posted: **Fri Feb 06, 2009 11:29 am**

Hi to all,

I would like to know
what concentration of standard solution should be used to perform the wavelength accuracy test with aqueous caffeine solution for HPLC-UV-Vis detector calibration and is there any guideline to use caffeine as standard.

Thanks in advance

Nageshwar

Posted: **Fri Feb 06, 2009 4:20 pm**

I would favour the use of a STD which does not produce an absorbance in excess of 1000 mAU (1 AU) at the theoretical caffeine absorbance maximum.

We use a 25 ÂµL injection of a 50 Âµg/mL STD solution.

Posted: **Sat Feb 07, 2009 7:34 pm**

you should check if your detector has a mercury lamp.
this lamp has a set of very specific WL and it is used these days in many detector for WL accuracy of the optics

Posted: **Mon Feb 09, 2009 2:46 am**

Hi,

The detector is built with duterium lamp.

Thanks

Nageshwar