Many methods call for reconstituting experimental samples in methanol/ water, but diluting the standards with some sort of plasma (typically blank bovine plasma). Why not dilute the standards in methanol/ water to match the experimental samples?
because everyone is quantifying hormones extracted from plasma-based samples.
The choice of diluent for preparing standards and experimental samples in analytical methods can depend on several factors, including the solubility of the analyte, compatibility with the analysis technique, and the desired matrix for calibration.
Here are some reasons why diluting standards with plasma or other biological matrices may be preferred over diluting them with methanol/water to match the experimental samples:
It's important to note that the choice of diluent depends on the specific analytical method, the analyte being measured, and the overall objective of the analysis. Methanol/water dilution may still be appropriate for some cases where the analyte is stable and soluble in that solvent mixture and if matrix effects are not a significant concern.
Ultimately, the decision on the diluent should be made based on method requirements, validation considerations, and the desired accuracy and reliability of the results. It is recommended to consult the specific method or assay protocol, as well as relevant literature or guidelines, for guidance on appropriate dilution procedures for your specific analysis.
Because LC-MS is used for the quantitation of biological molecules found in blood plasma in very low concentrations (pg/mL, like Vitamin D).
The reason why standards are often diluted in plasma rather than methanol/water is to account for matrix effects. Matrix effects refer to the interference of sample components with the ionization and detection of analytes. such as proteins and lipids, which can affect the ionization efficiency and detection of analytes. By diluting standards in plasma, it better mimics the sample matrix, allowing for more accurate calibration and assessment of the analyte's concentration in real samples.
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