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Phosphate Buffer (pH 5.8 to 7.4) Preparation and Recipe

A simple phosphate buffer is used ubiquitously in biological experiments, as it can be adapted to a variety of pH levels, including isotonic. This wide range is due to phosphoric acid having 3 dissociation constants, (known in chemistry as a triprotic acid) allowing for formulation of buffers near each of the pH levels of 2.15, 6.86, or 12.32. Phosphate buffer is highly water soluble and has a high buffering capacity, but will inhibit enzymatic activity and precipitates in ethanol. The buffer is one of the most popular currently used, and is commonly employed in molecular and cell biology, chemistry, and material science, among many others.

To prepare  L of Phosphate Buffer (pH 5.8 to 7.4):
Change the value in the textbox above to scale the recipe volume

Current molarity:
0.1 M
Adjust the molarity of the solution by using the slider below
Current pH:
Adjust the pH of the solution by using the slider below

Table 1. Required components
Sodium Phosphate Dibasic Heptahydrate (mw: 268.07 g/mol)20.214 g0.07541 M
Sodium Phosphate Monobasic Monohydrate (mw: 137.99 g/mol)3.394 g0.02459 M
  1. Prepare 800 mL of distilled water in a suitable container.
  2. Add 20.214 g of Sodium Phosphate Dibasic Heptahydrate to the solution.
  3. Add 3.394 g of Sodium Phosphate Monobasic Monohydrate to the solution.
  4. Adjust solution to final desired pH using HCl or NaOH
  5. Add distilled water until the volume is 1 L.

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Physiological Buffer
pH Buffering
Sample Preparation
Cell/Culture/Growth Media
Gel Electrophoresis


This online tool may be cited as follows


"Quest Calculate™ Phosphate Buffer (pH 5.8 to 7.4) Preparation and Recipe." AAT Bioquest, Inc.1 Oct2022


AAT Bioquest, Inc. (2022October 1). Quest Calculate™ Phosphate Buffer (pH 5.8 to 7.4) Preparation and Recipe. AAT Bioquest.

This online tool has been cited in 2 publications, including

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Journal: (2021)
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Journal: Heliyon (2022): e09099