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I try to determine sulbactam in human plasma by hplc. there
are analytical mathods described sush as protein precipitation(
ACN,Methanol,ZnSO4) Liquid-liquid extraction (diethylether) and
derivatization. The most interesting one is protein precipitated
(ACN) plasma 1 ml+ACN 1 mlvortex 1 min and centrifuged 14000 rpm 45
min . but I still have overcome extraction problem , sample was not
clean blankplasma show many endogeneous peak in chromatogram. I can't
separate sulbactam peak from interferance from metabolic product, lack
of sensitivity. If someone has experience or idea to point me that
would be great , Thanks so much in advance.
siriporn chawanond.
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Hi!
I think your protein ppt is not complete.Have you got clear layer
after cetrifugation?? if not then this unprecipitated protein that may
cause interferance in your analysis(blank plasma).
Why don't you try 10% or 20% perchloric acid or trichloroacetic acid,
add only 25ul in 500ul plasma that would give you clear layer after
centrifugation.Centrifuge it at room temp.
Let me know if you overcome this problem by this method.
With Regds
Yogesh Patil
Research Scientist
Department of Bioequivalence & Clinical Research
Alembic Pharmaceuticals Ltd
Baroda-390003
INDIA
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