by
carls » Sat Sep 11, 2010 4:21 pm
I just go up to 2 ul or do I go up and down few times and then slowly go to 3 ul
I follow the above but I dispense the extra volume with the syringe still in contact with the sample/liquid. If you remove the syringe and expel the extra, how do you remove the extra?
There are several schools of thought (we are analytical chermists after all) regarding which technique gives the most reproducible results and I'll mention a few below.
1) Pull the needle out of the solution and dispense the extra volume with the needle in contact with wall of sample vessel. The only question here is, do you lose additional sample due to capillary action/surface tension.
2) With the needle still in the solution, dispense the extra volume then remove the needle from the solution. The question here is, does additional liquid remain on the needle.
Additonally, after getting the desired volume in the syringe some propose to pull up a few uL of air (so as to not remove sample thru adsorption onto the wipe) and wipe off the needle.
3) A further variation (probably way more than you wanted to know at this point) is to flush the syringe with clean solvent then pull a microliter of solvent into the syringe. Then pull a microliter or 2 of air followed by the desired volume of sample. You can pull the sample up a few times to rinse the syringe, just dont expell all liquid, i.e. leave some of the air and all of the clean solvent in the syringe. This variation, some argue, ensures better precision by rinsing the sample out of the syringe during the injection process and reducing variability due to evaporation from the needle while in the injection port (time dependant). Just be sure the total volume of solvent (clean + sample) doesn't exceed the vapor volume of your inlet liner.
Either way, as long as you follow one of the techniques consistently your results should be consistent.