Instructions for the Calibration of the Spectronic 20 GENESYS Spectronic 20 GENESYS Spectrophotometer (Source: Manufacturer's Instruction Manual)

Initial Preparation Make sure the cell holder in the sample compartment is empty before turning on the spectrophotometer. Turn the power switch (located at the rear lower left side) to ON ("I") Allow the spectrophotmeter to warm up for 30 minutes. (Scroll down for additional information.)

Making Measurements Press the "A/T/C" pad to select the desired mode (A, for Absorbance) Press the "nm (up)" or "nm (down)" pad to set the proper wavelength. [Holding down on the desired key changes the wavelength more quickly.] Insert the blank or reference solution into the cell holder in the sample compartment andclose the door. Press the "0 ABS/100%T" pad to set the blank or reference. The spectrophotometer is now calibrated for the chosen wavelength setting. This procedure must be repeated whenever the wavelength is changed. Remove the blank and insert the sample into the cell holder. Close the sample compartment door. The measurement for the sample will appear on the LCD display.

Preparation of Samples in Cuvets Spectrophotometric measurements are very sensitive and precise. By this, we meant that the instrument will reliably measure the absorbance of whatever solution is presented to it. Of course, if solutions are improperly prepared orpresented, the measurements will be in error. Adhering to the following procedures will ensure that your work will yield accurate results. Use the same cuvet for all measurements unless you have a set of precisely matched cuvets (ask yourself: "why?"). Rinse the cuvet twice with a small amount (~1mL) of solution that is to be measured. [Do not use water.] "Roll" the solution in the cuvet so all the inside surface get rinsed. Empty all rinses into a beaker that is beingused as a waste container.

Always measure the %T of the solution with the lowest concentration first then progress to the highest concentration (ask yourself: "why?"). Empty solutions from the cuvet into the waste container. Cuvets need to be about 3/4 full. Make sure there are no bubbles in the solution to be measured. [Bubbles diffract light!] Clean the outside of the cuvet with an absorbsent, lint-free, soft tissue such as KimWipes or Accuwipes. [Fingerprints diffract light!] Insert the cuvet into the cell holder the same way each time. If your cuvets have only two clear faces, be sure that the light beam will pass through them! NEVER THROW AWAY ANY SOLUTIONS UNTIL THE STANDARD CURVE IS DRAWN AND DETERMINED TO BE ACCURATE AND PRECISE. It is a lot quicker to re-measure solutions than to remake all of them.