Is it necessary to construct the calibration curve of the drug in plasma for in-vivo pharmacokinetic study? Can we use simple calibration curve of drug constructed in solvent not in the plasma matrix, for quantification of the drug in plasma samples?
A calibration curve serves to standardize activities and reactions and compare the known with the unknown. To draw an objective conclusion, there must be some reference to some standard.
To establish the calibration curve in a matrix similar to your unknown samples, in this case your matrix is plasma, is considered the best option to account for matrix interference with your analytes of interest. It is known as "Matrix-Matched Calibration". It is a common practice when you expect an intereference that may decrease or increase the analytes' signal in the Mass Spectrometer.
To establish the calibration curve in a matrix similar to your unknown samples, in this case your matrix is plasma, is considered the best option to account for matrix interference with your analytes of interest. It is known as "Matrix-Matched Calibration". It is a common practice when you expect an interference that may decrease or increase the analytes' signal in the Mass Spectrometer.
Thus, to answer your question in simple words, yes it is necessary, at least from analytical point of view.
You may refer to some publications about matrix effect in mass spectrometry, like on of my papers in my profile.
Please look at the following below links which may help you in your analysis:
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3520453/pdf/JZUSB13-0990.pdf
https://www.scielo.br/pdf/bjps/v53n2/2175-9790-bjps-53-02-e16047.pdf
Thanks
Best practice would be to prepare calibration standards in plasma (i.e. serially dilute in 100% matrix), then apply the same minimum required dilution to your standards as you do to your samples in assay diluent. For example with a protein tested by ELISA, if your on-plate standard range is 1000 ng/mL to 1 ng/mL, and your minimum required dilution is 10, you would prepare standards at 10000 ng/mL to 10 ng/mL in plasma, then dilute 10-fold in assay diluent and your samples 10-fold in assay diluent before plating.
For the purpose of comparing unknown values of evaluated parameters, and for objective validation of data, it is necessary in most cases to construct a calibration curve.
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