Phosphate buffer solution preparation
Phosphate Buffer:
Solutions from pH 5.8 to pH 8.0 may be prepared by mixing 50 mL of 0.2M potassium dihydrogen orthophosphate with the quantities of 0.2M sodium hydroxide specified in the following table and diluting to 200 mL with water.At 20° the solutions may be used as solutions of standard pH.
Phosphate Buffer pH 3.0:
Dissolve 34 g of potassium dihydrogen orthophosphate in 250 mL of water and adjust the pH of the solution to 3.0 with orthophosphoric acid.
Phosphate Buffer pH 3.5:
Dissolve 68 g of potassium dihydrogen orthophosphate in 1000 mL of water and adjust the pH of the solution to 3.5 with orthophosphoric acid.
Phosphate Buffer pH 4.0:
Dissolve 6.8 g of potassium dihydrogen orthophosphate in 700 mL of water, adjust the pH, if necessary, with a 10% v/v solution of orthophosphoric acid and add sufficient water to produce 1000 mL.
Phosphate Buffer pH 4.0, Mixed:
Dissolve 5.04 g of disodium hydrogen orthophosphate and 3.01 g of potassium dihydrogen orthophosphate in sufficient water to produce 1000 mL and adjust the pH with glacial acetic acid.
Phosphate Buffer pH 4.75:
Dilute 100 mL of 0.5M potassium dihydrogen orthophosphate to 800 mL with water, adjust to pH 4.75 with 0.1M sodium hydroxide and dilute to 1000 mL with water.Phosphate Buffer pH 4.9:
Dissolve 40 g of sodium dihydrogen orthophosphate and 1.2 g of sodium hydroxide in sufficient water to produce 100 mL. If necessary, adjust the pH with 1M sulfuric acid or 1M sodium hydroxide as required.Phosphate Buffer pH 5.4, Mixed:
Dissolve 1.76 g of disodium hydrogen orthophosphate and 13.61 g of potassium dihydrogen orthophosphate in sufficient water to produce 1000 mL. Adjust the pH with 0.05M orthophosphoric acid, if necessary.
Phosphate Buffer pH 6.8, Mixed:
Dissolve 28.80 g of disodium hydrogen orthophosphate and 11.45 g of potassium dihydrogen orthophosphate in sufficient water to produce 1000 mL.
Phosphate Buffer pH 6.8, 0.2M Mixed:
Mix 51 mL of a 2.72% w/v solution of potassium dihydrogen orthophosphate with 49 mL of a 7.16% w/v solution of disodium hydrogen orthophosphate and adjust the pH if necessary.
Phosphate Buffer pH 7.0, Mixed:
Dissolve 0.50 g of anhydrous disodium hydrogen orthophosphate and 0.301 g of potassium dihydrogen orthophosphate in sufficient water to produce 1000 mL.
Phosphate Buffer pH 7.0, 0.067M Mixed:
Dissolve 0.908 g of potassium dihydrogen phosphate in water and dilute to 100.0 mL with the same solvent (solution A).
Dissolve 2.38 g of disodium hydrogen phosphate in water and dilute to 100.0 mL with the same solvent (solution B).
Mix 38.9 mL of solution A and 61.1 mL of solution B. Adjust the pH if necessary.
Phosphate Buffer pH 7.0, 0.1M Mixed:
Dissolve 1.361 g of potassium dihydrogen phosphate in water and dilute to 100.0 mL with the same solvent. Adjust the pH using a 35 g/L solution of disodium hydrogen phosphate.
Phosphate Buffer pH 7.5, 0.2M:
Dissolve 27.22 g of potassium dihydrogen phosphate R in 930 mL of water, adjust to pH 7.5 (2.2.3) with a 300 g/L solution of potassium hydroxide and dilute to 1000.0 mL with water.
Phosphate Buffer pH 10, Mixed:
To 100 mL of 0.2M disodium hydrogen orthophosphate add 6.0 mL of 0.25M trisodium orthophosphate.
Phosphate Buffer, 0.025M Standard:
Dissolve 3.40 g of potassium dihydrogen orthophosphate and 3.55 g of anhydrous disodium hydrogen orthophosphate, both previously dried at 110° to 130° for 2 hours, in sufficient water to produce 1000 mL.
Phosphate buffer solution pH 2.0:
Dissolve 8.95 g of disodium hydrogen phosphate and 3.40 g of potassium dihydrogen phosphate in water and dilute to 1000.0 mL with the same solvent. If necessary adjust the pH with phosphoric acid.
Phosphate Buffer Solution pH 2.5, 0.2M:
Dissolve 27.2 g of potassium dihydrogen phosphate in about 900 mL of water, adjust to pH 2.5 with phosphoric acid and dilute to 1000 mL with water.
Phosphate Buffer Solution pH 2.8:
Dissolve 7.8 g of sodium dihydrogen phosphate in 900 mL of water, adjust to pH 2.8 with phosphoric acid and dilute to 1000 mL with the same solvent.
Phosphate Buffer Solution pH 3.0:
Mix 0.7 mL of phosphoric acid with 100 mL of water. Dilute to 900 mL with the same solvent. Adjust to pH 3.0 with strong sodium hydroxide solution and dilute to 1000 mL with water.
Phosphate Buffer Solution pH 3.0, 0.1M :
Dissolve 12.0 g of anhydrous sodium dihydrogen phosphate in water, adjust the pH with dilute phosphoric acid and dilute to 1000 mL with water.Phosphate Buffer Solution pH 3.0 :
Dissolve 3.40 g of potassium dihydrogen phosphate in 900 mL of water. Adjust to pH 3.0 with phosphoric acid and dilute to 1000.0 mL with water.
Phosphate Buffer Solution pH 3.2 :
To 900 mL of a 4 g/L solution of sodium dihydrogen phosphate, add 100 mL of a 2.5 g/L solution of phosphoric acid. Adjust the pH if necessary.
Phosphate Buffer Solution pH 3.2 :
Adjust a 35.8 g/L solution of disodium hydrogen phosphate to pH 3.2 with dilute phosphoric acid. Dilute 100.0 mL of the solution to 2000.0 mL with water.
Phosphate Buffer Solution pH 3.25 :
Dissolve 1.36 g of potassium dihydrogen orthophosphate in 1000 mL of water and adjust the pH of the solution to 3.25 ± 0.05 with dilute orthophosphoric acid. Filter through a membrane filter (nominal pore size 0.45 µm or finer).
Phosphate Buffer Solution pH 3.5 :
Dissolve 68.0 g of potassium dihydrogen phosphate in water and dilute to 1000.0 mL with the same solvent. Adjust the pH with phosphoric acid.
Phosphate Buffer Solution pH 4.5, 0.05M :
Dissolve 6.80 g of potassium dihydrogen phosphate in 1000.0 mL of water. The pH of the solution is 4.5.Phosphate Buffer Solution pH 5.0 :
Dissolve 2.72 g of potassium dihydrogen phosphate in 800 mL of water. Adjust the pH with 1 M potassium hydroxide and dilute to 1000 mL with water.
Phosphate Buffer Solution pH 5.4, 0.067M
Mix appropriate volumes of a 23.99 g/l solution of disodium hydrogen phosphate with a 9.12 g/L solution of sodium dihydrogen phosphate monohydrate to obtain pH 5.4.
Phosphate Buffer Solution pH 5.5 :
Dissolve 13.61 g of potassium dihydrogen phosphate in water and dilute to 1000.0 mL with the same solvent (solution A).
Dissolve 35.81 g of disodium hydrogen phosphate in water and dilute to 1000.0 mL with the same solvent (solution B).
Mix 96.4 mL of solution A and 3.6 mL of solution B.
Phosphate–Citrate Buffer Solution pH 5.5 :
Mix 56.85 mL of a 28.4 g/L solution of anhydrous disodium hydrogen phosphate and 43.15 mL of a 21 g/L solution of citric acid.
Phosphate Buffer Solution pH 5.6 :
Dissolve 0.908 g of potassium dihydrogen phosphate in water and dilute to 100.0 mL with the same solvent (solution A).
Dissolve 1.161 g of dipotassium hydrogen phosphate in water and dilute to 100.0 mL with the same solvent (solution B).
Mix 94.4 mL of solution A and 5.6 mL of solution B. If necessary, adjust to pH 5.6 using solution A or solution B.
Phosphate Buffer Solution pH 5.8 :
Dissolve 1.19 g of disodium hydrogen phosphate dihydrate and 8.25 g of potassium dihydrogen phosphate in water and dilute to 1000.0 mL with the same solvent.
Phosphate Buffer Solution pH 6.0 :
Mix 63.2 mL of a 71.5 g/L solution of disodium hydrogen phosphate and 36.8 mL of a 21 g/L solution of citric acid.
Phosphate Buffer Solution pH 6.0 R1:
Dissolve 6.8 g of sodium dihydrogen phosphate in water and dilute to 1000.0 mL with water. Adjust the pH with strong sodium hydroxide solution.
Phosphate Buffer Solution pH 6.0 R2 :
To 250.0 mL of 0.2 M potassium dihydrogen phosphate add 28.5 mL of 0.2 M sodium hydroxide and dilute to 1000.0 mL with water.
Phosphate Buffer Solution pH 6.3, 0.1M :
Dissolve 15.6 g of sodium dihydrogen orthophosphate in 900 mL of water, adjust the pH to 6.3 with 0.1M sodium hydroxide and add sufficient water to produce 1000 mL.
Phosphate Buffer Solution pH 6.4 :
Dissolve 2.5 g of disodium hydrogen phosphate, 2.5 g of sodium dihydrogen phosphate and 8.2 g of sodium chloride in 950 mL of water. Adjust the pH of the solution to 6.4 with 1 M sodium hydroxide or 1 M hydrochloric acid, if necessary. Dilute to 1000.0 mL with water.
Phosphate Buffer Solution pH 6.5 :
Dissolve 2.75 g of sodium dihydrogen phosphate and 4.5 g of sodium chloride in 500 mL of water. Adjust the pH with phosphate buffer solution pH 8.5.
Phosphate Buffer Solution pH 6.5, 0.1M :
Dissolve 13.80 g of sodium dihydrogen phosphate monohydrate in 900 mL of distilled water. Adjust the pH using a 400 g/L solution of sodium hydroxide. Dilute to 1000 mL with distilled water.
Phosphate Buffer Solution pH 6.7, 0.1M :
Dissolve 15.6 g of sodium dihydrogen phosphate in water and dilute to 1.0 L with the same solvent. Dissolve 17.8 g of disodium hydrogen phosphate dehydrate in water and dilute to 1.0 L with the same solvent. Mix the solutions, check the pH and if necessary adjust to pH 6.7.
Phosphate Buffer Solution pH 6.8 :
Mix 77.3 mL of a 71.5 g/L solution of disodium hydrogen phosphate with 22.7 mL of a 21 g/L solution of citric acid.
Phosphate Buffer Solution pH 6.8 R1 :
To 51.0 mL of a 27.2 g/L solution of potassium dihydrogen phosphate add 49.0 mL of a 71.6 g/L solution of disodium hydrogen phosphate. Adjust the pH if necessary.
Phosphate Buffer Solution pH 7.0 :
Mix 82.4 mL of a 71.5 g/L solution of disodium hydrogen phosphate with 17.6 mL of a 21 g/L solution of citric acid.
Phosphate Buffer Solution pH 7.0 R1 :
Mix 250.0 mL of 0.2 M potassium dihydrogen phosphate R and 148.2 mL of a 8 g/l solution of sodium hydroxide, adjust the pH if necessary. Dilute to 1000.0 mL with water.
Phosphate Buffer Solution pH 7.0 R2 :
Mix 50.0 mL of a 136 g/L solution of potassium dihydrogen phosphate with 29.5 mL of 1 M sodium hydroxide and dilute to 100.0 mL with water. Adjust the pH to 7.0.
Phosphate Buffer Solution pH 7.0 R3 :
Dissolve 5 g of potassium dihydrogen phosphate and 11 g of dipotassium hydrogen phosphate in 900 mL of water. Adjust to pH 7.0 with dilute phosphoric acid or dilute sodium hydroxide solution. Dilute to 1000 mL with water and mix.
Phosphate Buffer Solution pH 7.0 R4 :
Dissolve 28.4 g of anhydrous disodium hydrogen phosphate and 18.2 g of potassium dihydrogen phosphate in water and dilute to 500 mL with the same solvent.
Phosphate Buffer Solution pH 7.0 R5 :
Dissolve 28.4 g of anhydrous disodium hydrogen phosphate in 800 mL of water. Adjust the pH using a 30% m/m solution of phosphoric acid and dilute to 1000 mL with water.
Phosphate Buffer Solution pH 7.0, 0.025M :
Mix 1 volume of 0.063 M phosphate buffer solution pH 7.0 with 1.5 volumes of water.
Phosphate Buffer Solution pH 7.0, 0.03M :
Dissolve 5.2 g of dipotassium hydrogen phosphate in 900 mL of water for chromatography. Adjust the solution to pH 7.0 ± 0.1 using phosphoric acid and dilute to 1000 mL with water for chromatography.
Phosphate Buffer Solution pH 7.0, 0.05M
Mix 34 mL of water and 100 mL of 0.067 M phosphate buffer solution pH 7.0.
Phosphate Buffer Solution pH 7.0, 0.063M
Dissolve 5.18 g of anhydrous disodium hydrogen phosphate and 3.65 g of sodium dihydrogen phosphate monohydrate in 950 mL of water and adjust the pH with phosphoric acid; dilute to 1000.0 mL with water.Phosphate Buffer Solution pH 7.0, 0.067M
Dissolve 0.908 g of potassium dihydrogen phosphate R in water R and dilute to 100.0 mL with the same solvent (solution A). Dissolve 2.38 g of disodium hydrogen phosphate R in water R and dilute to 100.0 mL with the same solvent (solution B). Mix 38.9 mL of solution A and 61.1 mL of solution B. Adjust the pH if necessary.
Phosphate Buffer Solution pH 7.0, 0.1M
Dissolve 1.361 g of potassium dihydrogen phosphate in water and dilute to 100.0 mL with the same solvent. Adjust the pH using a 35 g/L solution of disodium hydrogen phosphate.
Phosphate Buffer Solution pH 7.2
Mix 87.0 mL of a 71.5 g/L solution of disodium hydrogen phosphate with 13.0 mL of a 21 g/L solution of citric acid.
Phosphate Buffer Solution pH 7.4
Add 250.0 mL of 0.2 M potassium dihydrogen phosphate to 393.4 mL of 0.1 M sodium hydroxide.
Phosphate Buffer Solution pH 7.5, 0.05M
Dissolve 0.89 g of disodium hydrogen phosphate dehydrate in about 80 mL of water . Adjust to pH 7.5 with an 8.5% v/v solution of phosphoric acid and dilute to 100.0 mL with water .
Phosphate Buffer Solution pH 7.5, 0.2M
Dissolve 27.22 g of potassium dihydrogen phosphate in 930 mL of water, adjust to pH 7.5 with a 300 g/L solution of potassium hydroxide and dilute to 1000.0 mL with water.
Phosphate Buffer Solution pH 7.5, 0.33M
Dissolve 119.31 g of disodium hydrogen phosphate in water and dilute to 1000.0 mL with the same solvent (solution A).
Dissolve 45.36 g of potassium dihydrogen phosphate in water and dilute to 1000.0 mL with the same solvent (solution B).
Mix 85 mL of solution A and 15 mL of solution B. Adjust the pH if necessary.
Phosphate Buffer Solution pH 8.0, 0.02M
To 50.0 mL of 0.2 M potassium dihydrogen phosphate add 46.8 mL of 0.2 M sodium hydroxide. Dilute to 500.0 mL with water.
Phosphate Buffer Solution pH 8.0, 0.1M
Dissolve 0.523 g of potassium dihydrogen phosphate and 16.73 g of dipotassium hydrogen phosphate in water and dilute to 1000.0 mL with the same solvent.
Phosphate buffer solution pH 8.0, 0.05 M
Dissolve 0.31 g of sodium dihydrogen phosphate in 70 mL of water and adjust to pH 8.0 with 1 M sodium hydroxide,then dilute to 100 mL with water.
Phosphate Buffer Solution pH 8.0, 1M
Dissolve 136.1 g of potassium dihydrogen phosphate in water, adjust the pH with 1 M sodium hydroxide. Dilute to 1000.0 mL with water.
Phosphate Buffer Solution pH 8.5
Dissolve 3.5 g of dipotassium hydrogen phosphate and 4.5 g of sodium chloride in 500 mL of water. Adjust the pH with a mixture of equal volumes of dilute phosphoric acid and water.
Potassium phosphate buffer solution pH 7.0
Dissolve 10 mg of bovine albumin and 68 mg of potassium dihydrogen in 30 mL of water. If necessary, adjust to pH 7.0 with potassium hydroxide. Dilute to 50 mL with water and filter.Saline pH 6.4, Phosphate-buffered
Dissolve 1.79 g of disodium hydrogen orthophosphate, 1.36 g of potassium dihydrogen orthophosphate and 7.02 g of sodium chloride in sufficient water to produce 1000 mL.
Saline pH 6.8, Phosphate-buffered
Dissolve 1.0 g of potassium dihydrogen phosphate, 2.0 g of dipotassium hydrogen phosphate and 8.5 g of sodium chloride in 900 mL of water, adjust the pH if necessary and dilute to 1000.0 mL with the same solvent.
Saline pH 7.2, Phosphate-albumin Buffered
Dissolve 10.75 g of disodium hydrogen phosphate, 7.6 g of sodium chloride and 10 g of bovine albumin in water and dilute to 1000.0 mL with the same solvent. Immediately before use adjust the pH using dilute sodium hydroxide solution or dilute phosphoric acid.
Saline pH 7.2 R1, Phosphate-albumin Buffered
Dissolve 10.75 g of disodium hydrogen phosphate, 7.6 g of sodium chloride and 1 g of bovine albumin in water and dilute to 1000.0 mL with the same solvent. Immediately before use adjust the pH using dilute sodium hydroxide solution or dilute phosphoric acid.
Saline pH 7.4, Phosphate-buffered
Dissolve 2.38 g of disodium hydrogen phosphate, 0.19 g of potassium dihydrogen phosphate and 8.0 g of sodium chloride in water. Dilute to 1000.0 mL with the same solvent. Adjust the pH if necessary.
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