I am Working about assay medicin in plasma with a UV Spectrophotometry but what I want to know if their a possibility to work on extract way or to work directly
the probleme i have i haven't any protocol to follow
Prepare plasma Sample subjected to ultrafiltration. The plasma and the ultrafiltrate samples is store at −20°C until analysis. Frozen plasma or ultrafiltrate samples are
thawed at room temperature and subjected to protein precipitation as follows. Samples aliquot of 200 μL and 20 μL of IS working solution were add to a 1.8 mL Eppendorf tube and the mixture was vortexe for 10 s. Then, 800 μL of acetonitrile was added and the mixture was vortexed for 1 min followed by centrifugation at 20,000 rpm for 5 min at 10°C. The supernatant was transferr into a clean glass tube and evaporate to dryness under a gentle nitrogen stream. The residue is reconstitute in 100 μL of mobile phase, vortexed for 1 min, centrifuge at 3000 rpm for 5 min, transferre into a plastic autosampler vial with pre‐slit septum, and 1 μL was injected into the UPLC MS/MS system.
And if u want to do with UV Spectrophotometry the follow this procedure:
The human plasma is dilute 1:10 with double distilled water.Appropriate aliquots from the stock solution and of the diluted human plasma to get the desired concentration of Standard Drug is pipetted in testing tubes and gently votex-mixed for 5 minutes. A blank plasma sample is also prepare, containing the amount of methanol used for the samples. The UV spectra of the solutions were thereafter plotted against double distilled water in the range 200-400 nm, with a slit of 2 nm, at 200 nm/min speed.
Previously, a background correction of the device was performed with water.
The zero-order spectra obtained were further derivatised (1-st to
fourth derivatives) using the facilities of the Mathematics mode
and used in the analysis.
Best of Luck
thank you a lot for your answer i want to know if there is a guidline to follow or those strategies are already proved by your application
Dear Colleague,
This protocol Already proved.According to Standard books.
UV spectrophotometric analysis may not suit for all plasma studies because of the sensitivity
Dear Akash
If it possible to tell me the names of thoes standard books.
Dear Habibur
I think we can avoid the probleme of sensitivity with a specifique optimal absorbance wavelength or the complementary absorbance wavelength.
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