Dansyl chloride is not very soluble and after SPE when is added in plasma extract generate a over pressure with dirty precolumn ...is it possible that precipitates with NaOH?
Hey Roberta Merlanti, nice to meet you!
We have published a paper for estradiol with Dansyl.
Article High Sensitivity Method Validated to Quantify Estradiol in H...
And, I remember that once I've developed a method for Propofol using Dansyl.
For estradiol, we performed a double liquid-liquid extraction.
For propofol I've used both simple ACN precipitation and a liquid-liquid. It depends how level-sensitivity equipment was avaiable.
Is mandatory to use SPE or could you think in a simpler approach?
My best regards!
we used 20ul of NaOH 100mM in the exctract with the presence of Dansyl cloride (total volume 120ul). We evaporeted and resospended with mobile phase (50ul)
It is possible that we damage the column ? Did you mean this? the pH range is 2-9 for poroshell C8
"We evaporeted and resospended with mobile phase (50ul)";
Regarded the addition of base only to samples: If you dried down and removed the base, dissolved the sample into the mobile phase (initial conditions), then you should be OK. - - *We have seen client's inject samples dissolved into pure base into their HPLC columns (NaOH) resulting in damage to the support and clogging.
Use clean fresh columns. FULLY dissolve all samples and filter them (0.45u) before injecting into any HPLC system. Use proper column washing procedures before and after each analysis. Follow good chromatography fundamentals.
Dear Andre I try your method for estradiol with a double liquid liquid extraction and dansyl cloride incubation but with my equipment (LTQ ion trap ESI source) on plasma was not succesfully.
Only after a single working session with dansyl cloride samples we have problems of high pressure..we wash the column and we used also SPE extraction (no liquid-liquid). We have problem with dansyl cloride from the beginning because it doesn't melt completly as reported in literature....
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