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- Posts: 22
- Joined: Thu Apr 25, 2013 9:21 am
We are doing a project on UPLC, and we have a "simple" question.
We are to calculate a final concentration from a measurement on UPLC as follows.
each sample:
5ul sample diluted in 50 ul buffer (1:10 dilution)
5ul 1:10 dilution in 100 ul precipitation solvent (1:20 dilution)
vortex + -20 degree precipitation for 15 min + spin 3000g for 10 min 4 degree
100 ul supernatant transfer to new tube and evaporate to dryness
Reconstitute with 25ul running solvent and load into UPLC
The concentrations are measured from a std. curve of pure anhydrous powder dissolved in running solvent and a 2-fold dilution series are made (concentration in uM)
However, the std samples are not prepped as the samples.
We hereafter interpolate "unknowns" from the std curve using graphed prism
We now just need to know if these steps are correct:
The final values for the samples are multiplied first by 10 and then 20 to get the final concentration in uM, as the std. curve ??
Thanks