Does anyone here have an established guideline on conducting HPLC on plasma sample? How much of standard solutions (containing serial concentration of drug) to be added into 500ul plasma prior analysis to obtain the standard curve?
adding 25 µl of standards, controls and saliva samples into appropriate
I have used the maximum volume 10 % in my work. I think this volume depend of extraction method, if you perform a protein precipitation extraction and the solvent solution it is the same, I think it is no problem for extraction.
If 5% spiking is to be done then 25 micro lit drug solution and if 10% spiking is to be done then 50 microliter drug solution is required in 500 microliter. % spiking depends on sensitivity of the method and efficiency of the extraction technique. If 5% spiking then standard drug solution should be 20 times higher concentrated and of 10% spiking then 10 times higher concentrated then the concentration required.
thanks so much for answering my question. Is there any reference based on your suggestions?
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