BMB 471 – Problem Set I Answers 1. Determining whether or not a spectrophotometer is accurately measuring absorbance is vital in obtaining reliable data in the laboratory. A researcher can make this determination by using a series of neutral density filters with known (true) absorbance values (A.U.). The accuracy of absorbance measurement of the spectrophotometer in question can be determined by comparing the values of known absorbance for each filter to the values of absorbance displayed when said filter is inserted into the machine. 2. Absorbance measurements of a chromophore by a spectrophotometer might be inaccurate due to a variety of real and apparent deviations. For instance, the chromophore’s observed absorbance value might deviate due to the analyte reacting with itself or other solutes, or due to a very high concentration of analyte (which damages the ability of the chromophore to absorb light). Also, the observed absorbance value of a chromophore might deviate due to stray light
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This note was uploaded on 03/19/2008 for the course BMB 471 taught by Professor Bowlby during the Spring '08 term at Michigan State University.