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HPLC Analysis of Blood Morphine Levels

HPLC Precipitation Sample Prep Blood Morphine

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#1 DCZach

DCZach

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Posted 28 November 2011 - 10:49 AM

I am trying to analyze blood morphine levels using liquid chromatography with UV detection. I plan on using a gradient flow (ammonium acetate to acetonitrile as the mobile phase) while observing wavelengths @254nm in ambient temperature with a C18 packed column and a guard column with comparable stats.

I need to process the blood samples at the outset. I am looking to use ammonium acetate as a protein precipitation agent but am unsure if this is possible. I have read on the forum that individuals use it to crash DNA but cannot seem to see anything about crash proteins as a whole out of the sample. If we can add in ammonium acetate we will then spin it down in SpinX vials (with a 0.2um pore) to remove lipids, proteins and excess matrices.

Does anyone know if using ammonium acetate would work to clean up a blood sample for HPLC analysis?

I also know that acetonitrile can be used but I worry what adding that to my sample followed by injection into the HPLC where the initial mobile phase will be ammonium acetate.

Thanks so much for your time-

Zach





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