4.1.3. Buffer solutions EUROPEAN PHARMACOPOEIA 9.

01/2017:40103 0.1 M Phosphate buffer solution pH 3.0. 4011500.

Dissolve 12.0 g of anhydrous sodium dihydrogen phosphate R
in water R, adjust the pH with dilute phosphoric acid R1 and
dilute to 1000 mL with water R.
Phosphate buffer solution pH 3.0. 4000500.
4.1.3. BUFFER SOLUTIONS
Mix 0.7 mL of phosphoric acid R with 100 mL of water R.
Buffered acetone solution. 4000100. Dilute to 900 mL with the same solvent. Adjust to pH 3.0 with
Dissolve 8.15 g of sodium acetate R and 42 g of sodium strong sodium hydroxide solution R and dilute to 1000 mL
chloride R in water R, add 68 mL of 0.1 M hydrochloric acid with water R.
and 150 mL of acetone R and dilute to 500 mL with water R. Phosphate buffer solution pH 3.0 R1. 4010000.
Buffer solution pH 2.0. 4000200. Dissolve 3.40 g of potassium dihydrogen phosphate R in
Dissolve 6.57 g of potassium chloride R in water R and add 900 mL of water R. Adjust to pH 3.0 with phosphoric acid R
119.0 mL of 0.1 M hydrochloric acid. Dilute to 1000.0 mL and dilute to 1000.0 mL with water R.
with water R. Phosphate buffer solution pH 3.2. 4008100.
0.125 M Phosphate buffer solution pH 2.0. 4015600. To 900 mL of a 4 g/L solution of sodium dihydrogen
Dissolve 17.0 g of potassium dihydrogen phosphate R and phosphate R, add 100 mL of a 2.5 g/L solution of phosphoric
17.8 g of anhydrous disodium hydrogen phosphate R in water R acid R. Adjust the pH if necessary.
and dilute to 1.0 L with the same solvent. If necessary adjust Phosphate buffer solution pH 3.2 R1. 4008500.
the pH with phosphoric acid R.
Adjust a 35.8 g/L solution of disodium hydrogen phosphate
Phosphate buffer solution pH 2.0. 4007900. dodecahydrate R to pH 3.2 with dilute phosphoric acid R.
Dilute 100.0 mL of the solution to 2000.0 mL with water R.
Dissolve 8.95 g of disodium hydrogen phosphate
dodecahydrate R and 3.40 g of potassium dihydrogen Phosphate buffer solution pH 3.25. 4014900.
phosphate R in water R and dilute to 1000.0 mL with the same
Dissolve about 1.36 g of potassium dihydrogen phosphate R in
solvent. If necessary adjust the pH with phosphoric acid R.
1000 mL of water R and adjust to pH 3.25 ± 0.05 with dilute
Sulfate buffer solution pH 2.0. 4008900. phosphoric acid R. Filter through a membrane filter (nominal
pore size 0.45 μm or finer).
Dissolve 132.1 g of ammonium sulfate R in water R and dilute
to 500.0 mL with the same solvent (Solution A). Carefully and Buffer solution pH 3.5. 4000600.
with constant cooling stir 14 mL of sulfuric acid R into about Dissolve 25.0 g of ammonium acetate R in 25 mL of water R
400 mL of water R ; allow to cool and dilute to 500.0 mL with and add 38.0 mL of hydrochloric acid R1. Adjust the
water R (Solution B). Mix equal volumes of solutions A and B. pH if necessary with dilute hydrochloric acid R or dilute
Adjust the pH if necessary. ammonia R1. Dilute to 100.0 mL with water R.
Buffer solution pH 2.2. 4010500. Phosphate buffer solution pH 3.5. 4000700.
Mix 6.7 mL of phosphoric acid R with 55.0 mL of a 40 g/L Dissolve 68.0 g of potassium dihydrogen phosphate R in
solution of sodium hydroxide R and dilute to 1000.0 mL with water R and dilute to 1000.0 mL with the same solvent. Adjust
water R. the pH with phosphoric acid R.
Buffer solution pH 2.5. 4000300. Buffer solution pH 3.6. 4000800.
Dissolve 100 g of potassium dihydrogen phosphate R in 800 mL To 250.0 mL of 0.2 M potassium hydrogen phthalate R add
of water R ; adjust to pH 2.5 with hydrochloric acid R and 11.94 mL of 0.2 M hydrochloric acid. Dilute to 1000.0 mL
dilute to 1000.0 mL with water R. with water R.
Buffer solution pH 2.5 R1. 4000400. Buffer solution pH 3.7. 4000900.
To 4.9 g of dilute phosphoric acid R add 250 mL of water R. To 15.0 mL of acetic acid R add 60 mL of ethanol (96 per
Adjust the pH with dilute sodium hydroxide solution R and cent) R and 20 mL of water R. Adjust to pH 3.7 by the addition
dilute to 500.0 mL with water R. of ammonia R. Dilute to 100.0 mL with water R.
0.2 M Phosphate buffer solution pH 2.5. 4014100. Buffered copper sulfate solution pH 4.0. 4001000.
Dissolve 27.2 g of potassium dihydrogen phosphate R in about Dissolve 0.25 g of copper sulfate pentahydrate R and 4.5 g
900 mL of water R, adjust to pH 2.5 with phosphoric acid R of ammonium acetate R in dilute acetic acid R and dilute to
and dilute to 1.0 L with water R. 100.0 mL with the same solvent.
Phosphate buffer solution pH 2.8. 4010600. 0.1 M Sodium acetate buffer solution pH 4.0. 4013800.
Dissolve 7.8 g of sodium dihydrogen phosphate R in 900 mL of Dissolve 822 mg of sodium acetate R in 100 mL of water R
water R, adjust to pH 2.8 with phosphoric acid R and dilute to (solution A). Dilute 1.44 mL of glacial acetic acid R in 250 mL
1000 mL with the same solvent. of water R (solution B). Titrate 100 mL of solution B using
about 20 mL of solution A.
Buffer solution pH 3.0. 4008000.
Dissolve 21.0 g of citric acid monohydrate R in 200 mL of 1 M Acetate buffer solution pH 4.4. 4001100.
sodium hydroxide and dilute to 1000 mL with water R. Dilute Dissolve 136 g of sodium acetate R and 77 g of ammonium
40.3 mL of this solution to 100.0 mL with 0.1 M hydrochloric acetate R in water R and dilute to 1000.0 mL with the same
acid. solvent ; add 250.0 mL of glacial acetic acid R and mix.
0.25 M Citrate buffer solution pH 3.0. 4012600. Phthalate buffer solution pH 4.4. 4001200.
Dissolve 5.3 g of citric acid monohydrate R in 80 mL of Dissolve 2.042 g of potassium hydrogen phthalate R in 50 mL
water R. Adjust the pH with 1 M sodium hydroxide and dilute of water R, add 7.5 mL of 0.2 M sodium hydroxide and dilute
to 100.0 mL with water R. to 200.0 mL with water R.

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EUROPEAN PHARMACOPOEIA 9.0 4.1.3. Buffer solutions

Acetate buffer solution pH 4.5. 4012500. 0.067 M Phosphate buffer solution pH 5.4. 4012000.
Dissolve 77.1 g of ammonium acetate R in water R. Add 70 mL Mix appropriate volumes of a 23.99 g/L solution of disodium
of glacial acetic acid R and dilute to 1000.0 mL with water R. hydrogen phosphate dodecahydrate R with a 9.12 g/L solution
of sodium dihydrogen phosphate monohydrate R to obtain
0.5 M Ammonium acetate buffer solution pH 4.5. 4014200. pH 5.4.
Mix 14.3 mL of glacial acetic acid R and 470 mL of water R
and adjust to pH 4.5 with concentrated ammonia R. Dilute to Acetate-edetate buffer solution pH 5.5. 4001900.
500.0 mL with water R. Dissolve 250 g of ammonium acetate R and 15 g sodium
0.05 M Phosphate buffer solution pH 4.5. 4009000. edetate R in 400 mL of water R and add 125 mL of glacial
acetic acid R.
Dissolve 6.80 g of potassium dihydrogen phosphate R in
1000.0 mL of water R. The pH of the solution is 4.5. Buffer solution pH 5.5. 4001800.
Sodium acetate buffer solution pH 4.5. 4010100. Dissolve 54.4 g of sodium acetate R in 50 mL of water R,
Dissolve 63 g of anhydrous sodium acetate R in water R, heating to 35 °C if necessary. After cooling, slowly add 10 mL
add 90 mL acetic acid R and adjust to pH 4.5, and dilute to of anhydrous acetic acid R. Shake and dilute to 100.0 mL with
1000 mL with water R. water R.

Acetate buffer solution pH 4.6. 4001400. Phosphate buffer solution pH 5.5. 4002000.
Dissolve 5.4 g of sodium acetate R in 50 mL of water R, add Dissolve 13.61 g of potassium dihydrogen phosphate R in water R
2.4 g of glacial acetic acid R and dilute to 100.0 mL with and dilute to 1000.0 mL with the same solvent (solution A).
water R. Adjust the pH if necessary. Dissolve 35.81 g of disodium hydrogen phosphate R in water R
and dilute to 1000.0 mL with the same solvent (solution B).
Succinate buffer solution pH 4.6. 4001500. Mix 96.4 mL of solution A and 3.6 mL of solution B.
Disssolve 11.8 g of succinic acid R in a mixture of 600 mL of
water R and 82 mL of 1 M sodium hydroxide and dilute to Phosphate-citrate buffer solution pH 5.5. 4008700.
1000.0 mL with water R. Mix 56.85 mL of a 28.4 g/L solution of anhydrous disodium
hydrogen phosphate R and 43.15 mL of a 21 g/L solution of
Acetate buffer solution pH 4.7. 4001600.
citric acid monohydrate R.
Dissolve 136.1 g of sodium acetate R in 500 mL of water R. Mix
250 mL of this solution with 250 mL of dilute acetic acid R. Phosphate buffer solution pH 5.6. 4011200.
Shake twice with a freshly prepared, filtered, 0.1 g/L solution of
Dissolve 0.908 g of potassium dihydrogen phosphate R
dithizone R in chloroform R. Shake with carbon tetrachloride R in water R and dilute to 100.0 mL with the same solvent
until the extract is colourless. Filter the aqueous layer to (solution A). Dissolve 1.161 g of dipotassium hydrogen
remove traces of carbon tetrachloride. phosphate R in water R and dilute to 100.0 mL with the same
Acetate buffer solution pH 4.7 R1. 4013600. 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
Dissolve 136.1 g of sodium acetate R in 500 mL of water R. Mix
or solution B.
250 mL of this solution with 250 mL of dilute acetic acid R.
Acetate buffer solution pH 5.0. 4009100. Phosphate buffer solution pH 5.8. 4002100.
To 120 mL of a 6 g/L solution of glacial acetic acid R add Dissolve 1.19 g of disodium hydrogen phosphate dihydrate R
100 mL of 0.1 M potassium hydroxide and about 250 mL of and 8.25 g of potassium dihydrogen phosphate R in water R
water R. Mix. Adjust the pH to 5.0 with a 6 g/L solution of and dilute to 1000.0 mL with the same solvent.
acetic acid R or with 0.1 M potassium hydroxide and dilute to
1000.0 mL with water R. Acetate buffer solution pH 6.0. 4002200.
Dissolve 100 g of ammonium acetate R in 300 mL of water R,
Citrate buffer solution pH 5.0. 4010700. add 4.1 mL of glacial acetic acid R, adjust the pH if necessary
Prepare a solution containing 20.1 g/L of citric acid using ammonia R or acetic acid R and dilute to 500.0 mL with
monohydrate R and 8.0 g/L of sodium hydroxide R. Adjust the water R.
pH with dilute hydrochloric acid R.
Diethylammonium phosphate buffer solution pH 6.0.
0.2 M Deuterated sodium phosphate buffer solution 4002300.
pH 5.0. 4013900.
Dilute 68 mL of phosphoric acid R to 500 mL with water R.
Dissolve 2.76 g of sodium dihydrogen phosphate monohydrate R To 25 mL of this solution add 450 mL of water R and 6 mL
in 90 mL of deuterium oxide R, adjust the pH with a deuterated of diethylamine R, adjust to pH 6 ± 0.05, if necessary, using
solution of phosphoric acid R or a deuterated 1 M solution of diethylamine R or phosphoric acid R and dilute to 500.0 mL
sodium hydroxide R, dilute to 100 mL with deuterium oxide R with water R.
and mix.
Phosphate buffer solution pH 6.0. 4002400.
Phosphate buffer solution pH 5.0. 4011300.
Mix 63.2 mL of a 71.5 g/L solution of disodium hydrogen
Dissolve 2.72 g of potassium dihydrogen phosphate R in 800 mL
phosphate dodecahydrate R and 36.8 mL of a 21 g/L solution
of water R. Adjust the pH with 1 M potassium hydroxide and
of citric acid monohydrate R.
dilute to 1000 mL with water R.
Sodium acetate buffer solution pH 5.0. 4015500. Phosphate buffer solution pH 6.0 R1. 4002500.
Dissolve 50.0 g of sodium acetate R in 10.0 mL of glacial Dissolve 6.8 g of sodium dihydrogen phosphate R in water R
acetic acid R and add water R. Adjust to pH 5.0 with a 4.2 g/L and dilute to 1000.0 mL with water R. Adjust the pH with
solution of sodium hydroxide R or with glacial acetic acid R strong sodium hydroxide solution R.
and dilute to 1000.0 mL with water R.
Phosphate buffer solution pH 6.0 R2. 4002600.
Buffer solution pH 5.2. 4001700. To 250.0 mL of 0.2 M potassium dihydrogen phosphate R add
Dissolve 1.02 g of potassium hydrogen phthalate R in 30.0 mL 28.5 mL of 0.2 M sodium hydroxide and dilute to 1000.0 mL
of 0.1 M sodium hydroxide. Dilute to 100.0 mL with water R. with water R.

General Notices (1) apply to all monographs and other texts 561
4.1.3. Buffer solutions EUROPEAN PHARMACOPOEIA 9.0

Phosphate buffer solution pH 6.4. 4002800. containing 7.8 g/L of sodium dihydrogen phosphate R and
Dissolve 2.5 g of disodium hydrogen phosphate dodecahydrate R, 23 g/L of sodium chloride R to adjust the pH. Dissolve in the
2.5 g of sodium dihydrogen phosphate R and 8.2 g of sodium solution sufficient sodium azide R to give a 0.2 g/L solution.
chloride R in 950 mL of water R. Adjust the pH of the solution
to 6.4 with 1 M sodium hydroxide or 1 M hydrochloric acid, if Maleate buffer solution pH 7.0. 4003600.
necessary. Dilute to 1000.0 mL with water R. Dissolve 10.0 g of sodium chloride R, 6.06 g of
tris(hydroxymethyl)aminomethane R and 4.90 g of maleic
0.5 M Phthalate buffer solution pH 6.4. 4009200. anhydride R in 900 mL of water R. Adjust the pH using a
Dissolve 100 g of potassium hydrogen phthalate R in water R 170 g/L solution of sodium hydroxide R. Dilute to 1000.0 mL
and dilute to 1000.0 mL with the same solvent. Adjust the pH with water R.
if necessary, using strong sodium hydroxide solution R. Storage : at 2 °C to 8 °C ; use within 3 days.
Buffer solution pH 6.5. 4002900.
0.025 M Phosphate buffer solution pH 7.0. 4009400.
Dissolve 60.5 g of disodium hydrogen phosphate
dodecahydrate R and 46 g of potassium dihydrogen phosphate R Mix 1 volume of 0.063 M phosphate buffer solution pH 7.0 R
in water R. Add 100 mL of 0.02 M sodium edetate and 20 mg with 1.5 volumes of water R.
of mercuric chloride R and dilute to 1000.0 mL with water R. 0.03 M Phosphate buffer solution pH 7.0. 4010300.
Imidazole buffer solution pH 6.5. 4003000. Dissolve 5.2 g of dipotassium hydrogen phosphate R in
Dissolve 6.81 g of imidazole R, 1.23 g of magnesium sulfate R 900 mL of water for chromatography R. Adjust the solution to
and 0.73 g of calcium sulfate R in 752 mL of 0.1 M hydrochloric pH 7.0 ± 0.1 using phosphoric acid R and dilute to 1000 mL
acid. Adjust the pH if necessary and dilute to 1000.0 mL with with water for chromatography R.
water R.
0.05 M Phosphate buffer solution pH 7.0. 4012400.
0.1 M phosphate buffer solution pH 6.5. 4010800. Mix 34 mL of water R and 100 mL of 0.067 M phosphate buffer
Dissolve 13.80 g of sodium dihydrogen phosphate solution pH 7.0 R.
monohydrate R in 900 mL of distilled water R. Adjust the pH
using a 400 g/L solution of sodium hydroxide R. Dilute to 0.063 M Phosphate buffer solution pH 7.0. 4009500.
1000 mL with distilled water R. Dissolve 5.18 g of anhydrous disodium hydrogen phosphate R
Phosphate buffer solution pH 6.5. 4012800. and 3.65 g of sodium dihydrogen phosphate monohydrate R in
950 mL of water R and adjust the pH with phosphoric acid R ;
Dissolve 2.75 g of sodium dihydrogen phosphate R and 4.5 g of dilute to 1000.0 mL with water R.
sodium chloride R in 500 mL of water R. Adjust the pH with
phosphate buffer solution pH 8.5 R. 0.067 M Phosphate buffer solution pH 7.0. 4003800.
Buffer solution pH 6.6. 4003100. Dissolve 0.908 g of potassium dihydrogen phosphate R
To 250.0 mL of 0.2 M potassium dihydrogen phosphate R add in water R and dilute to 100.0 mL with the same solvent
89.0 mL of 0.2 M sodium hydroxide. Dilute to 1000.0 mL with (solution A). Dissolve 2.38 g of disodium hydrogen phosphate
water R. dodecahydrate R in water R and dilute to 100.0 mL with the
same solvent (solution B). Mix 38.9 mL of solution A and
0.1 M Phosphate buffer solution pH 6.7. 4014300. 61.1 mL of solution B. Adjust the pH if necessary.
Dissolve 15.6 g of sodium dihydrogen phosphate R in water R
and dilute to 1.0 L with the same solvent. Dissolve 17.8 g of 0.1 M Phosphate buffer solution pH 7.0. 4008200.
disodium hydrogen phosphate dihydrate R in water R and Dissolve 1.361 g of potassium dihydrogen phosphate R in
dilute to 1.0 L with the same solvent. Mix the solutions, check water R and dilute to 100.0 mL with the same solvent. Adjust
the pH and if necessary adjust to pH 6.7. the pH using a 35 g/L solution of disodium hydrogen phosphate
dodecahydrate R.
Phosphate buffered saline pH 6.8. 4003200.
Dissolve 1.0 g of potassium dihydrogen phosphate R, 2.0 g Phosphate buffer solution pH 7.0. 4003700.
of dipotassium hydrogen phosphate R and 8.5 g of sodium Mix 82.4 mL of a 71.5 g/L solution of disodium hydrogen
chloride R in 900 mL of water R, adjust the pH if necessary phosphate dodecahydrate R with 17.6 mL of a 21 g/L solution
and dilute to 1000.0 mL with the same solvent. of citric acid monohydrate R.
Phosphate buffer solution pH 6.8. 4003300. Phosphate buffer solution pH 7.0 R1. 4003900.
Mix 77.3 mL of a 71.5 g/L solution of disodium hydrogen Mix 250.0 mL of 0.2 M potassium dihydrogen phosphate R and
phosphate dodecahydrate R with 22.7 mL of a 21 g/L solution 148.2 mL of a 8 g/L solution of sodium hydroxide R, adjust the
of citric acid monohydrate R. pH if necessary. Dilute to 1000.0 mL with water R.
Phosphate buffer solution pH 6.8 R1. 4003400.
Phosphate buffer solution pH 7.0 R2. 4004000.
To 51.0 mL of a 27.2 g/L solution of potassium dihydrogen
phosphate R add 49.0 mL of a 71.6 g/L solution of disodium Mix 50.0 mL of a 136 g/L solution of potassium dihydrogen
hydrogen phosphate dodecahydrate R. Adjust the pH if phosphate R with 29.5 mL of 1 M sodium hydroxide and dilute
necessary. to 100.0 mL with water R. Adjust the pH to 7.0 ± 0.1.
Storage : at 2 °C to 8 °C. Phosphate buffer solution pH 7.0 R3. 4008600.
1 M tris-hydrochloride buffer solution pH 6.8. 4009300. Dissolve 5 g of potassium dihydrogen phosphate R and 11 g
Dissolve 60.6 g of tris(hydroxymethyl)aminomethane R in of dipotassium hydrogen phosphate R in 900 mL of water R.
400 mL of water R. Adjust the pH with hydrochloric acid R Adjust to pH 7.0 with dilute phosphoric acid R or dilute sodium
and dilute to 500.0 mL with water R. hydroxide solution R. Dilute to 1000 mL with water R and mix.

Buffer solution pH 7.0. 4003500. Phosphate buffer solution pH 7.0 R4. 4010200.
To 1000 mL of a solution containing 18 g/L of disodium Dissolve 28.4 g of anhydrous disodium hydrogen phosphate R
hydrogen phosphate dodecahydrate R and 23 g/L of sodium and 18.2 g of potassium dihydrogen phosphate R in water R
chloride R add sufficient (about 280 mL) of a solution and dilute to 500 mL with the same solvent.

562 See the information section on general monographs (cover pages)

EUROPEAN PHARMACOPOEIA 9.0 4.1.3. Buffer solutions

Phosphate buffer solution pH 7.0 R5. 4011400. Buffer solution pH 7.4. 4004600.
Dissolve 28.4 g of anhydrous disodium hydrogen phosphate R Dissolve 0.6 g of potassium dihydrogen phosphate R, 6.4 g of
in 800 mL of water R. Adjust the pH using a 30 per cent m/m disodium hydrogen phosphate dodecahydrate R and 5.85 g of
solution of phosphoric acid R and dilute to 1000 mL with sodium chloride R in water R, and dilute to 1000.0 mL with the
water R. same solvent. Adjust the pH if necessary.
Phosphate buffer solution pH 7.0 R6. 4015300. Phosphate buffered saline pH 7.4. 4005000.
Dissolve 3.56 g of disodium hydrogen phosphate dihydrate R Dissolve 2.38 g of disodium hydrogen phosphate
in 950 mL of water for chromatography R. Adjust the pH dodecahydrate R, 0.19 g of potassium dihydrogen phosphate R
with phosphoric acid R and dilute to 1.0 L with water for and 8.0 g of sodium chloride R in water. Dilute to 1000.0 mL
chromatography R. with the same solvent. Adjust the pH if necessary.
Phosphate buffer solution pH 7.0 R7. 4015700. Phosphate buffer solution pH 7.4. 4004800.
Dissolve 35 g of dipotassium hydrogen phosphate R in 900 mL Add 250.0 mL of 0.2 M potassium dihydrogen phosphate R to
of water R, adjust to pH 7.0 with dilute phosphoric acid R and 393.4 mL of 0.1 M sodium hydroxide.
dilute to 1.0 L with water R.
Tris(hydroxymethyl)aminomethane buffer solution pH 7.4.
Potassium phosphate buffer solution pH 7.0. 4014700. 4012100.
Dissolve 10 mg of bovine albumin R and 68 mg of potassium Dissolve 30.3 g of tris(hydroxymethyl)aminomethane R in
dihydrogen phosphate R in 30 mL of water R. If necessary, approximately 200 mL of water R. Add 183 mL of 1 M
adjust to pH 7.0 with potassium hydroxide R. Dilute to 50 mL hydrochloric acid. Dilute to 500.0 mL with water R. Note :
with water R and filter. the pH is 7.7-7.8 at room temperature and 7.4 at 37 °C. This
Sodium/calcium acetate buffer solution pH 7.0. 4014800. solution is stable for several months at 4 °C.
Dissolve 10 mg of bovine albumin R and 32 mg of calcium Tris(hydroxymethyl)aminomethane sodium chloride
acetate R in 60 mL of water R. Add 580 μL of glacial acetic buffer solution pH 7.4. 4004900.
acid R and adjust to pH 7.0 with 2 M sodium hydroxide R.
Dilute to 100 mL with water R and filter. Dissolve 6.08 g of tris(hydroxymethyl)aminomethane R, 8.77 g
of sodium chloride R in 500 mL of distilled water R. Add 10.0 g
Tetrabutylammonium buffer solution pH 7.0. 4010900. of bovine albumin R. Adjust the pH using hydrochloric acid R.
Dissolve 6.16 g of ammonium acetate R in a mixture of 15 mL Dilute to 1000.0 mL with distilled water R.
of tetrabutylammonium hydroxide solution (400 g/L) R and
Tris(hydroxymethyl)aminomethane sodium chloride
185 mL of water R. Adjust the pH with nitric acid R.
buffer solution pH 7.4 R1. 4012200.
Buffered salt solution pH 7.2. 4004300. Dissolve 0.1 g of bovine albumin R in a mixture containing
Dissolve in water R 8.0 g of sodium chloride R, 0.2 g of 2 mL of tris(hydroxymethyl)aminomethane buffer solution
potassium chloride R, 0.1 g of anhydrous calcium chloride R, pH 7.4 R and 50 mL of a 5.84 mg/mL solution of sodium
0.1 g of magnesium chloride R, 3.18 g of disodium hydrogen chloride R. Dilute to 100.0 mL with water R.
phosphate dodecahydrate R and 0.2 g of potassium dihydrogen
phosphate R and dilute to 1000.0 mL with water R. Tris-sodium acetate buffer solution pH 7.4. 4012900.
Dissolve 6.3 g of tris(hydroxymethyl)aminomethane R and
Buffer solution pH 7.2. 4004100. 4.9 g of anhydrous sodium acetate R in 900 mL of water R.
To 250.0 mL of 0.2 M potassium dihydrogen phosphate R add Adjust to pH 7.4 with sulfuric acid R and dilute to 1000 mL
175.0 mL of 0.2 M sodium hydroxide. Dilute to 1000.0 mL with water R.
with water R. Adjust the pH if necessary.
Tris-sodium acetate-sodium chloride buffer solution
Phosphate-albumin buffered saline pH 7.2. 4004400. pH 7.4. 4013000.
Dissolve 10.75 g of disodium hydrogen phosphate Dissolve 30.0 g of tris(hydroxymethyl)aminomethane R, 14.5 g
dodecahydrate R, 7.6 g of sodium chloride R and 10 g of bovine of anhydrous sodium acetate R and 14.6 g of sodium chloride R
albumin R in water R and dilute to 1000.0 mL with the same in 900 mL of water R. Add 0.50 g of bovine albumin R. Adjust
solvent. Immediately before use adjust the pH using dilute to pH 7.4 with sulfuric acid R and dilute to 1000 mL with
sodium hydroxide solution R or dilute phosphoric acid R. water R.
Phosphate-albumin buffered saline pH 7.2 R1. 4009600. Borate buffer solution pH 7.5. 4005200.
Dissolve 10.75 g of disodium hydrogen phosphate Dissolve 2.5 g of sodium chloride R, 2.85 g of disodium
dodecahydrate R, 7.6 g of sodium chloride R and 1 g of bovine tetraborate R and 10.5 g of boric acid R in water R and dilute to
albumin R in water R and dilute to 1000.0 mL with the same 1000.0 mL with the same solvent. Adjust the pH if necessary.
solvent. Immediately before use adjust the pH using dilute
sodium hydroxide solution R or dilute phosphoric acid R. Storage : at 2 °C to 8 °C.

Phosphate buffer solution pH 7.2. 4004200. Buffer (HEPES) solution pH 7.5. 4009700.
Mix 87.0 mL of a 71.5 g/L solution of disodium hydrogen Dissolve 2.38 g of HEPES R in about 90 mL of water R. Adjust
phosphate dodecahydrate R with 13.0 mL of a 21 g/L solution the pH to 7.5 with sodium hydroxide solution R. Dilute to
of citric acid monohydrate R. 100 mL with water R.

Imidazole buffer solution pH 7.3. 4004500. 0.05 M Phosphate buffer solution pH 7.5. 4014400.
Dissolve 3.4 g of imidazole R and 5.8 g of sodium chloride R in Dissolve 0.89 g of disodium hydrogen phosphate dihydrate R
water R, add 18.6 mL of 1 M hydrochloric acid and dilute to in about 80 mL of water R. Adjust to pH 7.5 with an 8.5 per
1000.0 mL with water R. Adjust the pH if necessary. cent V/V solution of phosphoric acid R and dilute to 100.0 mL
with water R.
Barbital buffer solution pH 7.4. 4004700.
Mix 50 mL of a solution in water R containing 19.44 g/L of 0.2 M Phosphate buffer solution pH 7.5. 4005400.
sodium acetate R and 29.46 g/L of barbital sodium R with Dissolve 27.22 g of potassium dihydrogen phosphate R in
50.5 mL of 0.1 M hydrochloric acid, add 20 mL of an 85 g/L of 930 mL of water R, adjust to pH 7.5 with a 300 g/L solution of
sodium chloride R and dilute to 250 mL with water R. potassium hydroxide R and dilute to 1000.0 mL with water R.

General Notices (1) apply to all monographs and other texts 563
4.1.3. Buffer solutions EUROPEAN PHARMACOPOEIA 9.0

0.33 M Phosphate buffer solution pH 7.5. 4005300. Tris-sodium acetate buffer solution pH 8.0. 4013100.
Dissolve 119.31 g of disodium hydrogen phosphate Dissolve 6.3 g of tris(hydroxymethyl)aminomethane R and
dodecahydrate R in water R and dilute to 1000.0 mL with 4.9 g of anhydrous sodium acetate R in 900 mL of water R.
the same solvent (solution A). Dissolve 45.36 g of potassium Adjust to pH 8.0 with sulfuric acid R and dilute to 1000 mL
dihydrogen phosphate R in water R and dilute to 1000.0 mL with water R.
with the same solvent (solution B). Mix 85 mL of solution A
and 15 mL of solution B. Adjust the pH if necessary. Tris-sodium acetate-sodium chloride buffer solution
pH 8.0. 4013200.
0.05 M Tris-hydrochloride buffer solution pH 7.5. 4005600.
Dissolve 30.0 g of tris(hydroxymethyl)aminomethane R, 14.5 g
Dissolve 6.057 g of tris(hydroxymethyl)aminomethane R in of anhydrous sodium acetate R and 14.6 g of sodium chloride R
water R and adjust the pH with hydrochloric acid R. Dilute to in 900 mL of water R. Add 0.50 g of bovine albumin R. Adjust
1000.0 mL with water R. to pH 8.0 with sulfuric acid R and dilute to 1000 mL with
1 M Tris-hydrochloride buffer solution pH 7.5. 4014500. water R.
Dissolve 12.11 g of tris(hydroxymethyl)aminomethane R in Tris(hydroxymethyl)aminomethane buffer solution pH 8.1.
90 mL of water R, adjust to pH 7.5 with hydrochloric acid R 4006200.
and dilute to 100.0 mL with water R.
Dissolve 0.294 g of calcium chloride R in 40 mL of
Tris(hydroxymethyl)aminomethane buffer solution pH 7.5. tris(hydroxymethyl)aminomethane solution R and adjust the
4005500. pH with 1 M hydrochloric acid. Dilute to 100.0 mL with
Dissolve 7.27 g of tris(hydroxymethyl)aminomethane R and water R.
5.27 g of sodium chloride R in water R, and adjust the pH if
necessary. Dilute to 1000.0 mL with water R. Tris-glycine buffer solution pH 8.3. 4006300.
Dissolve 6.0 g of tris(hydroxymethyl)aminomethane R and
Sodium citrate buffer solution pH 7.8 (0.034 M sodium
28.8 g of glycine R in water R and dilute to 1000.0 mL with the
citrate, 0.101 M sodium chloride). 4009800.
same solvent. Dilute 1 volume to 10 volumes with water R
Dissolve 10.0 g of sodium citrate R and 5.90 g of sodium immediately before use.
chloride R in 900 mL of water R. Adjust the pH by addition of
hydrochloric acid R and dilute to 1000 mL with water R. Tris-hydrochloride buffer solution pH 8.3. 4011800.
0.0015 M Borate buffer solution pH 8.0. 4006000. Dissolve 9.0 g of tris(hydroxymethyl)aminomethane R in 2.9 L
Dissolve 0.572 g of disodium tetraborate R and 2.94 g of of water R. Adjust the pH with 1 M hydrochloric acid. Adjust
calcium chloride R in 800 mL of water R. Adjust the pH with the volume to 3 L with water R.
1 M hydrochloric acid. Dilute to 1000.0 mL with water R.
0.05 M Tris-hydrochloride buffer solution pH 9.0. 4013500.
Buffer solution pH 8.0. 4005900. Dissolve 0.605 g of tris(hydroxymethyl)aminomethane R in
To 50.0 mL of 0.2 M potassium dihydrogen phosphate R add water R. Adjust the pH with 1 M hydrochloric acid and dilute
46.8 mL of 0.2 M sodium hydroxide. Dilute to 200.0 mL with to 100.0 mL with water R.
water R.
Barbital buffer solution pH 8.4. 4006400.
Buffer solution pH 8.0 R1. 4010400.
Dissolve 8.25 g of barbital sodium R in water R and dilute to
Dissolve 20 g of dipotassium hydrogen phosphate R in 900 mL 1000.0 mL with the same solvent.
of water R. Adjust the pH with phosphoric acid R. Dilute to
1000 mL with water R. Tris-EDTA BSA buffer solution pH 8.4. 4006500.
0.02 M Phosphate buffer solution pH 8.0. 4006100. Dissolve 6.1 g of tris(hydroxymethyl)aminomethane R, 2.8 g
To 50.0 mL of 0.2 M potassium dihydrogen phosphate R add of sodium edetate R, 10.2 g of sodium chloride R and 10 g
46.8 mL of 0.2 M sodium hydroxide. Dilute to 500.0 mL with of bovine albumin R in water R, adjust to pH 8.4 using 1 M
water R. hydrochloric acid and dilute to 1000.0 mL with water R.

0.02 M Sodium phosphate buffer solution pH 8.0. 4013700. Tris(hydroxymethyl)aminomethane-EDTA buffer solution
Dissolve 0.31 g of sodium dihydrogen phosphate R in 70 mL of pH 8.4. 4006600.
water R and adjust to pH 8.0 with 1 M sodium hydroxide, then Dissolve 5.12 g of sodium chloride R, 3.03 g of
dilute to 100 mL with water R. tris(hydroxymethyl)aminomethane R and 1.40 g of sodium
edetate R in 250 mL of distilled water R. Adjust the pH to 8.4
0.1 M Phosphate buffer solution pH 8.0. 4008400. using hydrochloric acid R. Dilute to 500.0 mL with distilled
Dissolve 0.523 g of potassium dihydrogen phosphate R and water R.
16.73 g of dipotassium hydrogen phosphate R in water R and
dilute to 1000.0 mL with the same solvent. Tris(hydroxymethyl)aminomethane-EDTA buffer solution
pH 8.4 R1. 4015100.
1 M Phosphate buffer solution pH 8.0. 4007800.
Dissolve 10.20 g of sodium chloride R, 6.10 g of
Dissolve 136.1 g of potassium dihydrogen phosphate R in
tris(hydroxymethyl)aminomethane R, 2.80 g of sodium
water R, adjust the pH with 1 M sodium hydroxide. Dilute to
edetate R and 1.00 g of macrogol 6000 R or 2.00 g of bovine
1000.0 mL with water R.
albumin R or of human albumin R in 800 mL of water R.
1 M Tris-hydrochloride buffer solution pH 8.0. 4012700. Adjust to pH 8.4 with hydrochloric acid R and dilute to 1.0 L
Dissolve 121.1 g of tris(hydroxymethyl)aminomethane R and with water R.
1.47 g of calcium chloride R in 900 mL of water R. Adjust the
Guanidine-tris(hydroxymethyl)aminomethane-EDTA
pH with hydrochloric acid R and dilute to 1000.0 mL with
buffer solution pH 8.5. 4014600.
water R.
Dissolve 1.0 g of sodium edetate R, 12.1 g of
Tris-hydrochloride buffer solution pH 8.0. 4012300. tris(hydroxymethyl)aminomethane R and 57.0 g of
Dissolve 1.21 g of tris(hydroxymethyl)aminomethane R and guanidine hydrochloride R in 35 mL of water R. Adjust to
29.4 mg of calcium chloride R in water R. Adjust the pH with pH 8.5 with hydrochloric acid R and dilute to 100 mL with
1 M hydrochloric acid and dilute to 100.0 mL with water R. water R.

564 See the information section on general monographs (cover pages)

EUROPEAN PHARMACOPOEIA 9.0 4.2.1. Primary standards for volumetric solutions

Phosphate buffer solution pH 8.5. 4013300. Ammonium chloride buffer solution pH 10.4. 4011000.
Dissolve 3.5 g of dipotassium hydrogen phosphate R and 4.5 g Dissolve 70 g of ammonium chloride R in 200 mL of water R,
of sodium chloride R in 500 mL of water R. Adjust the pH add 330 mL of concentrated ammonia R and dilute to
with a mixture of equal volumes of dilute phosphoric acid R 1000.0 mL with water R. If necessary, adjust to pH 10.4 with
and water R. ammonia R.

Tris acetate buffer solution pH 8.5. 4006700. Borate buffer solution pH 10.4. 4011100.
Dissolve 24.64 g of boric acid R in 900 mL of distilled water R.
Dissolve 0.294 g of calcium chloride R and 12.11 g of Adjust the pH using a 400 g/L solution of sodium hydroxide R.
tris(hydroxymethyl)aminomethane R in water R. Adjust the Dilute to 1000 mL with distilled water R.
pH with acetic acid R. Dilute to 1000.0 mL with water R.
Ammonium chloride buffer solution pH 10.7. 4013400.
Barbital buffer solution pH 8.6 R1. 4006900. Dissolve 67.5 g of ammonium chloride R in water R, add
Dissolve in water R 1.38 g of barbital R, 8.76 g of barbital 570 mL of concentrated ammonia R and dilute to 1000.0 mL
sodium R and 0.38 g of calcium lactate pentahydrate R and with water R.
dilute to 1000.0 mL with the same solvent.
Buffer solution pH 10.9. 4007600.
1.5 M Tris-hydrochloride buffer solution pH 8.8. 4009900. Dissolve 6.75 g of ammonium chloride R in ammonia R and
Dissolve 90.8 g of tris(hydroxymethyl)aminomethane R in dilute to 100.0 mL with the same solvent.
400 mL of water R. Adjust the pH with hydrochloric acid R Total-ionic-strength-adjustment buffer. 4007700.
and dilute to 500.0 mL with water R. Dissolve 58.5 g of sodium chloride R, 57.0 mL of glacial
acetic acid R, 61.5 g of sodium acetate R and 5.0 g of
3 M Tris-hydrochloride buffer solution pH 8.8. 4015000.
cyclohexylenedinitrilotetra-acetic acid R in water R and dilute
Dissolve 363.3 g of tris(hydroxymethyl)aminomethane R in to 500.0 mL with the same solvent. Adjust to pH 5.0 to 5.5
500 mL of water R. Adjust the pH with hydrochloric acid R with a 335 g/L solution of sodium hydroxide R and dilute to
and dilute to 1 L with water R. 1000.0 mL with distilled water R.
Buffer (phosphate) solution pH 9.0. 4008300. Total-ionic-strength-adjustment buffer R1. 4008800.
Dissolve 1.74 g of potassium dihydrogen phosphate R in 80 mL Dissolve 210 g of citric acid monohydrate R in 400 mL
of water R, adjust the pH with 1 M potassium hydroxide and of distilled water R. Adjust to pH 7.0 with concentrated
dilute to 100.0 mL with water R. ammonia R. Dilute to 1000.0 mL with distilled water R
(solution A). Dissolve 132 g of ammonium phosphate R
Buffer solution pH 9.0. 4007000. in distilled water R and dilute to 1000.0 mL with the
Dissolve 6.18 g of boric acid R in 0.1 M potassium chloride R same solvent (solution B). To a suspension of 292 g of
and dilute to 1000.0 mL with the same solvent. Mix 1000.0 mL (ethylenedinitrilo)tetra-acetic acid R in about 500 mL
of this solution and 420.0 mL of 0.1 M sodium hydroxide. of distilled water R, add about 200 mL of concentrated
ammonia R to dissolve. Adjust the pH to 6 to 7 with
Buffer solution pH 9.0 R1. 4007100. concentrated ammonia R. Dilute to 1000.0 mL with distilled
water R (solution C). Mix equal volumes of solution A, B,
Dissolve 6.20 g of boric acid R in 500 mL of water R and adjust and C and adjust to pH 7.5 with concentrated ammonia R.
the pH with 1 M sodium hydroxide (about 41.5 mL). Dilute to
1000.0 mL with water R. Buffer solution pH 11. 4014000.
Dissolve 6.21 g of boric acid R, 4.00 g of sodium hydroxide R
Tris(hydroxymethyl)aminomethane buffer solution pH 9.0. and 3.70 g of potassium chloride R in 500 mL of water R and
4015200. dilute to 1000 mL with the same solvent.
Dissolve 1.21 g of tris(hydroxymethyl)aminomethane R in
0.1 M Phosphate buffer solution pH 11.3. 4015400.
950 mL of water for chromatography R. Adjust to pH 9.0
with acetic acid R and dilute to 1000.0 mL with water for Dissolve 17.4 g of dipotassium hydrogen phosphate R in about
chromatography R. 950 mL of water R, adjust to pH 11.3 using a 100 g/L solution
of potassium hydroxide R and dilute to 1.0 L with water R.
Ammonium chloride buffer solution pH 9.5. 4007200. Filter through a membrane filter (nominal pore size 0.45 μm).
Dissolve 33.5 g of ammonium chloride R in 150 mL of water R,
add 42.0 mL of concentrated ammonia R and dilute to 4.2. VOLUMETRIC ANALYSIS
250.0 mL with water R.
Storage : in a polyethylene container. 04/2010:40201

Ammonium chloride buffer solution pH 10.0. 4007300.

Dissolve 5.4 g of ammonium chloride R in 20 mL of water R,
add 35.0 mL of ammonia R and dilute to 100.0 mL with
water R. 4.2.1. PRIMARY STANDARDS FOR
VOLUMETRIC SOLUTIONS
Diethanolamine buffer solution pH 10.0. 4007500.
Primary standards for volumetric solutions are indicated
Dissolve 96.4 g of diethanolamine R in water R and dilute by the suffix RV. Primary standards of suitable quality may
to 400 mL with the same solvent. Add 0.5 mL of an 186 g/L be obtained from commercial sources or prepared by the
solution of magnesium chloride R and adjust the pH with 1 M following methods.
hydrochloric acid. Dilute to 500.0 mL with water R.
Arsenious trioxide. As2O3. (Mr 197.8). 2000100. [1327-53-3].
0.1 M Ammonium carbonate buffer solution pH 10.3. Sublime arsenious trioxide R in a suitable apparatus.
4011900.
Storage : over anhydrous silica gel R.
Dissolve 7.91 g of ammonium carbonate R in 800 mL
of water R. Adjust the pH with dilute sodium hydroxide Benzoic acid. C7H6O2. (Mr 122.1). 2000200. [65-85-0].
solution R. Dilute to 1000.0 mL with water R. Sublime benzoic acid R in a suitable apparatus.

General Notices (1) apply to all monographs and other texts 565