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How to Make a Cadenza Buffer

Cadenza buffers are commonly used in chromatography experiments to limit interference and eliminate background staining. It is a relatively safe substance, although it does pose a risk of respiratory, skin and eye irritation. Instructors of honors chemistry students or college professors can aid their students' understanding of balancing chemical equations and base properties by assigning students to create a Cadenza buffer from raw materials. A license to purchase controlled chemicals and access to a laboratory facility with proper safety equipment is required.

Things You'll Need

Sodium chloride
Liquid polyethylene glycol
Tris(hydroxymethyl) aminomethane
Tris(hydroxymethyl) aminomethane hydrochloride
4 graduated cylinders
Beaker
Distilled water
Glass stirring rod
Glass jar with lid

Instructions

- 1
  Check the laboratory fire extinguisher, safety shower and eyewash station to ensure they are all in working order in case of a spill during the experiment. Don neoprene gloves and safety goggles prior to working with buffer-solution chemicals.
- 2
  Prepare the separate components of the buffer solution by adding 150 mg of sodium chloride to a graduated cylinder. Pour 40 mL polyethylene glycol into a second graduated cylinder. Prepare the two remaining cylinders with 55 mg Tris(hydroxymethyl) aminomethane and 150 mg Tris(hydroxymethyl) aminomethane hydrochloride, respectively.
- 3
  Pour sodium chloride into a beaker. Add the Tris(hydroxymethyl) aminomethane and Tris(hydroxymethyl) aminomethane hydrochloride and mix the dry ingredients with a glass stirring rod. Slowly add the polyethylene glycol while continuing to stir.
- 4
  Add 20 mL distilled water to help the solid chemicals dissolve. Stir with a glass stirring rod until the solution is thoroughly mixed.
- 5
  Perform a pH test to check the neutrality of the Cadenza buffer. The solution should indicate a pH level near 7.0.
- 6
  Store the Cadenza buffer in a glass container with an airtight lid to prevent evaporation.

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