HPLC Information and Resources

HPLC Protocols - including fluorescence detection
Supplies Companies
Column Care
HPLC systems

Companies  

Alltech (or here for non-Java version, probably less prone to crashing):  For pricing info (Online Store), use account CA095.  But don't order online.
Vydac

Column care

Storage conditions:  >50% organic in H2O (NOT BUFFER).  Absolutely no TFA.

Exposure of column to pH6.5 or greater:  Use silica saturation column between pump and injector, to minimize loss of silica from column (and resulting void volume and decrease in resolution).

From Western Analytical

From Alltech

From Vydac

HPLC Systems

Agilent Technologies (formerly Hewlett-Packard)

HPLC Protocols

Fluorescence detection with PE 650-10 S relic.

The PE 650-10 S is dead.

Power up sequence - (make sure the HPLC is off before lighting the xenon lamp!)

1. Fluorimeter on
2. Xenon power supply on
3. Lamp start on power supply (make sure the HPLC is off or the power surge will cause an error on the LC which can be reversed only by cycling the main power switch on the back of the LC.)
4. HP interface 35900E on
5. HPLC power on
6. HPLC diode array on
7. Start software/load method
8. Pump on

Hardware/software details

Fluorimeter

Make sure the flow cell flows from bottom right to top left, when looking from front to back of instrument along excitation beam path.  If the flow is from bottom left to top right, flip the flow cell upside-down.  I know, it doesn't make sense, but otherwise you will probably get bubbles trapped in the flow cell.  If the flow is from top to bottom:  get with the program, you don't know what you're doing!

Initial suggested slitwidths are 10 nm.

Software

You need to make sure the Instrument/Setup ADC Signals/Stop Datatime setting is the same as the end of the run time.