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With traditional methodologies that rely on standard fixed-pathlength UV-visible spectroscopy, the turn-around time may be on the order of hours.That is attributed to the need for careful sample handling and preparation, particularly in creating the dilutions necessary for bringing samples into the linear range of an instrument’s calibration.Additionally, the error created when performing those dilutions can have a significant impact on the calculated sample concentration.Dimensional equality then allows for replacement of the left-hand side of the second equation above with the slope term from the third equation, yielding the following: .That resulting equation is referred to as the slope spectroscopy equation.It may be used to calculate a sample’s concentration — if the molar absorption coefficient is known — by dividing it into the slope as such: .Because pathlength selection is computer controlled and optimized based on the absorbance achieved, the above approach ensures rapid, accurate, and reproducible results.The Solo VPE spectroscopy system is equipped with a highly precise and repeatable computer-controlled linear stage that can quickly ( values.
Background and Purpose: The biopharmaceutical industry’s need for fast, accurate concentration measurements of protein-containing products is of prime importance.
Commonly referred to as “A testing, the speed and turn-around time of results are nearly as critical as accuracy because manufacturing personnel are usually waiting for those data before they can continue processing.
No longer are scientists bound to the time-consuming, error-prone use of dilution factors and fixed-pathlength measurements in determining the concentration of an analyte in solution.
Using the slope spectroscopy technique, the Solo VPE system (from C Technologies) offers a new method of determining analyte concentration based on the Beer–Lambert law and slope derived from absorbance measurements made at multiple pathlengths (1).
Mathematics: The Beer–Lambert law is expressed as , where A is the measured absorbance, α is the molar absorption coefficient, l is the pathlength, and c is the sample concentration.
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This equation can then be rearranged for use with slope spectroscopy:, where m is the slope of the regression line, and b is the y-intercept.