Hello sir.
I am currently a master's student, and I am conducting pharmacokinetic studies in rats on the drug Posaconazole. However, I have encountered some problems recently. That is, the calibration curve of Posaconazole drug through HPLC analysis in rat urine is not drawn. It was well drawn in Rat plasma.
The pretreatment method of Posaconazole drug for HPLC analysis on the rat plasma that I did is, 50ul of rat plasma sample, 100ul of Acetonitrile and 1ul of Internal Standard (10mg/ml concentration), and after 3min of vortexing and 10min of 13.0g centrifuge, then take 80ul of supernatant for HPLC analysis. It was put in a vial and analyzed.
The same was done for Rat Urine, but the analysis is not working well. I need your desperate help. If you have any additional information you need, please tell me and I will let you know. Thank you.
Hi
You're method cant work when the different matrices are used. It means even if you change the method from serum to blood plasma method may not give the same results. In your case, it two completely different matrices. Plasma pH and rat urine p His probably different (could be from 5.5-7.5) then you should use some buffer and try to stabilize the sample or generally think about how to stabilize pH and solubility in the sample. Please find a link with quite a nice example of work with urine preparation and TCA .
https://iris.uniroma1.it/retrieve/handle/11573/1194522/902925/Campestre_Analysis_2017.pdf
Best regards
Tomasz
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