HPLC Rookie questions

[Davidsonchemgrad]

Quick storyline behind this...

I just graduated college last August. The first of the year I was hired to work for a company and essentially be a guys understudy to learn how to run the HPLC and develop methods. Needless to say I understood the theory behind normal and reverse phase HPLC as well as a little about Ion Pairing, but I had very little actual experience. Long story short my boss got cancer and has since passed away. Now I am stuck with still very little experience and even less direction, but am supposed to be developing methods.

so...

First things first I am looking to take a course in HPLC method development. I have looked at the Waters course partly because we run a Waters instrument and partly because I have heard it is a great course. Any incite or recommendations?

Second I have some serious method development questions. I would like to be able to set up some methods for analysis of surfactants (I know this may be broad, but we use many) and common plasticizers (essentially the phthalates). I have very little direction at this point.

I have developed some less sophisticated methods for analysis of dye particles. One method simply uses 52% 0.1M Acetic Acid and 48% THF with .1 uL injections at 1.0mL/min. The second method is slightly more complex using 55% Ammonium Acetate/TBAH @ pH 7 and 45% THF. All samples are currently being dissolved in 90%THF/10%H2O. We use a Sunfire Column (5um, 4.6 x 100 mm, C18) and a PDA detector. The methods are isocratic 30 min runs at room temp. We also have an RI detector which I believe may be important in surfactant analysis, but we have not had to use the RI to this point.

Sorry if this is slightly unorganized. I can obviously give more details, but wanted to see if people had any incite first. Thanks in advance.

[chromatographer1]

No matter how good the course is, it is usually less than a week in length. There are limits in learning highly technical skills in such a short time, but by all means go and take note of who else is there and network network network.

There are some excellent books available concerning HPLC method development.

Use some of the petty cash fund and buy, read, and learn a lot.

It will be worth it.

best wishes,

Rod

[Consumer Products Guy]

Agilent has some free on-line courses in HPLC test method development and other subjects http://www.chem.agilent.com/Scripts/education.asp

Also, we only use THF here for HPLC separations when we really need its unique separation qualities, due to its odor, toxicity, and tendency to form peroxides. For dyes, maybe you can get by with preserved THF if your wavelength of detection is in the visible region....

[DR]

LCR's Systematic HPLC Method Development Using DryLab course, and the accompanying Snyder et al book (and a copy of Drylab) would combine to put you in a good place very quickly.

[juddc]

Echo on DR's comments.

I've only ever taken one LC course in my 15 years as a chromotagrapher; a very good one to train in qualifying instrumentation (IQ/OQ/PQ)...in 2005. The rest of it I've done through reading and experimentation, making mistakes and fixing them. I was never formally trained myself, but have trained a number of chemists successfully.

If I was in your position (again), I'd grab a copy of Snyder's book, read as many relevant applications as you can (bug column manufacturers to start), ask questions here, and don't be afraid to experiment on your own. As long as you follow some very basic rules (filter your mobile phase and samples, don't run a pH 12 buffer on a silica column support, and the like) you won't break anything, so have at it!

[tom jupille]

If you have six hours to spare, I'll put in a plug for the "live on the web" course we're doing with GMP Training Systems (three two-hour sessions, July 13, 20, and 27).

More details:
http://www.sepsci.com/chromforum/viewtopic.php?t=6232

[Uwe Neue]

I wrote a book chapter a few years ago about method development, at least about how to get started. Contact me and I send it to you!