method development

[uppala]

how to develop a HPLC method for a given compound?
intially, what parametres we look for given compound?

[Noser222]

Read a book about the basics of HPLC.

Not to be mean, but that's where you have to start.

[Jade.Barker]

1. I agree read a basic book. Try: "Modern HPLC for the Practicing Scientists" ISB# 978-0-471-72789-7

2. Then decide what data you need, and why-

What Data is needed
For example- Our laboratory tests fermentation samples. The different levels of the 8 components we test for show several things:
- The effectiveness of an enzyme for converting starch to sugar
- The probably bacterial levels based on acid production
- The efficiency of the yeast producing ethanol

Why the Data is needed
This data allows our technical consultants to make recommendations to the Ethanol Plants. For example a high acid sample may indicate a certain area of the plant should be treated with industrial hygiene or antibiotics.

3. Talk to a HPLC Vendor. When our lab was new we had a meeting with the Waters Ethanol HPLC specialist. He set up our first Method Set according to a Waters Method.

If you are new, it will be easier to start with an already existing method. Then as you run into problems, you can troubleshoot and adapt the method to fit your needs.

Good Luck!

[Bruce Hamilton]

Also, why not try to get on one of the courses/eseminars, including those available from this site.

If you plan on doing a lot of HPLC, then investiment in a good HPLC book , such as "Practical HPLC Method Development - 2nd Edition " by L.R.Snyder, J.J.Kirkland, J.L.Glajch 1997 ISBN 047100703X, is money well spent.

If your employer doesn't like buying books, just purchase it as a consumable amonst many other consumales from one of the many chemical and chromatography suppliers that offer such books.

Please keep having fun,

Bruce Hamilton

[Uwe Neue]

I can send you an article on our approach to method screening. I need your e-mail address for doing so.

[SIELC_Tech]

Here is our step-by-step guide based on the properties of your molecules:
http://www.sielc.com/MethodDevelopment_ ... dType.html

contact me if you have questions.

Vlad

[Jade.Barker]

Vlad,
Thats I looked at your link - thats a nice little tool. Thanks for posting it!