I am not sure whether anyone has much of an idea on using HPLC, but hope that someone does and can help me out a bit. I am trying to detect a number of compounds in a mixture using HPLC (currently developing a method), and have managed to separate and identify most on the HPLC. What I am not certain is whether the calibration curves could be prepared as mixtures, or whether I would need to prepare calibration curves for each of the compounds separately. Thank you

What detection system are you using (MS? UV? etc)? As long as all your compound peaks are fully resolved, there shouldn't be a problem running your cali plots using samples containing known quantities of a mixture of the compounds. Have you checked accuracy and precision yet? Usually worth running this part of the validation before doing the cali plot. Good luck :-)

======= Date Modified 18 Mar 2011 01:33:37 =======
I ran my calibration curve with a know amount of standard solution, made up of known amounts of pure compounds. Worked brilliantly! So using a mixture is no problem, as long as you know what's in there. And when in doubt you can always run a separate compound afterwards to confirm your conclusions.

On another note... I've been to a very good HPLC seminar in London last week... really helpful! Learned lots!

Thank you to both. Re the seminar. Who was it organised by? I am on the lookout for such seminars on HPLC and LC-MS method development, so would like to check them out. Thank you

The seminar I went to was organised by Thames Restek. They are doing a few all over the UK. I know that one of my other suppliers, Phenomenex, also does a series of seminars on HPLC. It's worth checking them out, because they are also nice networking opportunities and a way to get discounts from your suppliers ;)