Correction Baseline

I am developing a method for protein concentration in the Spectrophotometer. The concentration is measuring at 280 nm and correcting light scattering background at 320 nm.  I have noticed that the baseline is higher for this material varies from 0.1 to 0.8. I want to verify if I use the three point drop line form 320 nm to 340 nm  the baseline can be normalized? The concentration without correcting at 320 nm is 1.1 mg/mL when the subtraction of 320 nm is applied the concentration is 0.91 mg/mL.  This concentration was measured using SOLOVPE and it was 0.88 mg/ml. This concentration is similar to 0.91 mg/mL when the correction is applied.  I want to know if using a three point drop line it will be more efficient inserted of subtract a single wavelength of 320?

  • The use of a 3 point baseline probably won't make a considerable difference, but it's hard/impossible to say that with any certainty if you just look at a bunch of numbers collected from single wavelength measurements.  Instead, I would always recommend that you collect scans over the range of interest and look to see in the spectra if there is any variation.

Was this helpful?