preparing dilution of analyte [August 25, 2004]
Posted: Sun Aug 29, 2004 8:45 pm
By Lily Tan on Wednesday, August 25, 2004 - 10:13 pm:
I used to do Histology. In histology, we always prepare dilution of antibody in an eppendorf tube. Is it acceptable to prepare dilution of analyte in an eppendorf tube in HPLC? Otherwise, what sort of method should I use?
Is there any Good Practice for HPLC that I should take note as beginner? Any suggestions from anyone.
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By bert on Thursday, August 26, 2004 - 03:13 am:
It depends on the accuracy you want or need. For quantitative measurements, we always use volumetric flasks for the stock-solution as well as dilutions of the stock.
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By Ann on Thursday, August 26, 2004 - 08:03 am:
Hi Lily
I often prepare small volume of solutions in an eppendorf tube but I use Gilson variable volume pipettes to add the required volumes of stock solution and diluent to the tube. I would not use the printed graduations on the tubes.
Ann
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By HW Mueller on Thursday, August 26, 2004 - 11:49 pm:
Surely, thousands of people do what Ann described (me inclusive), but, if you get strange peaks remember that plastics may release monomers, softeners?, etc (never seen this in my work). Experienced "bio-people" told me (years ago when I started with bio-stuff) that many substances, especially proteins, have less tendency to stick to plastic than to glass. Of cause, as usual for the bio world, there are hydrophobic materials that would rather stick to plastic.
Now if you have organics (mobile phase....) in your solvent you should only let polyethylene devices (or teflon derivatives) get in contact. Also it is then better to use positive displacement pipets (piston in tip).
I used to do Histology. In histology, we always prepare dilution of antibody in an eppendorf tube. Is it acceptable to prepare dilution of analyte in an eppendorf tube in HPLC? Otherwise, what sort of method should I use?
Is there any Good Practice for HPLC that I should take note as beginner? Any suggestions from anyone.
-------------------------------------------------------------------------------------------------------
By bert on Thursday, August 26, 2004 - 03:13 am:
It depends on the accuracy you want or need. For quantitative measurements, we always use volumetric flasks for the stock-solution as well as dilutions of the stock.
-------------------------------------------------------------------------------------------------------
By Ann on Thursday, August 26, 2004 - 08:03 am:
Hi Lily
I often prepare small volume of solutions in an eppendorf tube but I use Gilson variable volume pipettes to add the required volumes of stock solution and diluent to the tube. I would not use the printed graduations on the tubes.
Ann
-------------------------------------------------------------------------------------------------------
By HW Mueller on Thursday, August 26, 2004 - 11:49 pm:
Surely, thousands of people do what Ann described (me inclusive), but, if you get strange peaks remember that plastics may release monomers, softeners?, etc (never seen this in my work). Experienced "bio-people" told me (years ago when I started with bio-stuff) that many substances, especially proteins, have less tendency to stick to plastic than to glass. Of cause, as usual for the bio world, there are hydrophobic materials that would rather stick to plastic.
Now if you have organics (mobile phase....) in your solvent you should only let polyethylene devices (or teflon derivatives) get in contact. Also it is then better to use positive displacement pipets (piston in tip).