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Buffer Essay

2459 words - 10 pages

Preparation of Buffers for Use in Enzyme Studies
(by G. Gomori)
The buffers described in this section are suitable for use either in enzymatic or histochemical
studies. The accuracy of the tables is within ± 0.05 pH at 23 ºC. In most cases the pH values
will not be off by more than ± 0.02 pH even at 37 ºC and at molarities slightly different from
those given (usually 0.05 M).
The methods of preparation described are not necessarily identical with those of the original
authors. The titration curves of the majority of the buffers recommended have been
redetermined by the writer. The buffers are arranged in the order of ascending pH range.
1. Hydrochloric Acid-Potassium Chloride Buffer.
Stocks ...view middle of the document...

1 M solution of sodium citrate (29.41 g C6H5O7Na3.2H2O in 1 L, the use of the
salt with 5½ H2O is not recommended).
x mL of A + y mL of B, diluted to a total of 100 mL
x
46.5
43.7
40.0
37.0
35.0
33.0
31.5
28.0
25.5
23.0
20.5
18.0
16.0
13.7
11.8
9.5
7.2

y
3.5
6.3
10.0
13.0
15.0
17.0
18.5
22.0
24.5
27.0
29.5
32.0
34.0
36.3
38.2
41.5
42.8

pH
3.0
3.2
3.4
3.6
3.8
4.0
4.2
4.4
4.6
4.8
5.0
5.2
5.4
5.6
5.8
6.0
6.2

6. Acetate Buffer.
Stocks solutions
A: 0.2 M solution of acetic acid (11.55 g in 1 L)
B: 0.2 M solution of sodium acetate (16.4 g C2H3O2Na or 27.2 g of C2H3O2Na.3H2O
in 1 L).
x mL of A + y mL of B, diluted to a total of 100 mL
x
46.3
44.0
41.0
36.8
30.5
25.5
20.0
14.8
10.5
8.8
4.8
jun 14, 2004

y
3.7
6.0
9.0
13.2
19.5
24.5
30.0
35.2
39.5
41.2
45.2

pH
3.6
3.8
4.0
4.2
4.4
4.6
4.8
5.0
5.2
5.4
5.6
Pag.3

7. Citrate-Phosphate Buffer.
Stocks solutions
A: 0.1 M solution of citric acid (19.21 g in 1 L)
B: 0.2 M solution of dibasic sodium phosphate (53.65 g of Na2HPO4.7H2O or 71.7 g
of Na2HPO4.12H2O in 1 L).
x mL of A + y mL of B, diluted to a total of 100 mL
x
44.6
42.2
39.8
37.7
35.9
33.9
32.3
30.7
29.4
27.8
26.7
25.2
24.3
23.3
22.2
21.0
19.7
17.9
16.9
15.4
13.6
9.1
6.5

y
5.4
7.8
10.2
12.3
14.1
16.1
17.7
19.3
20.6
22.2
23.3
24.8
25.7
26.7
27.8
29.0
30.3
32.1
33.1
34.6
36.4
40.9
43.6

pH
2.6
2.8
3.0
3.2
3.4
3.6
3.8
4.0
4.2
4.4
4.6
4.8
5.0
5.2
5.4
5.6
5.8
6.0
6.2
6.4
6.6
6.8
7.0

8. Succinate Buffer.
Stocks solutions
A: 0.2 M solution of succinic acid (23.6 g in 1L)
B: 0.2 M NaOH
25 mL of A + x mL of B, diluted to a total of 100 mL
x
7.5
10.0
13.3
16.7
20.0
23.5
jun 14, 2004

pH
3.8
4.0
4.2
4.4
4.6
4.8

x
26.7
30.3
34.2
37.5
40.7
43.5

pH
5.0
5.2
5.4
5.6
5.8
6.0
Pag.4

9. Phthalate-Sodium Hydroxide Buffer.
Stocks solutions
A: 0.2 M solution of potassium acid phthalate (40.84 g in 100 mL)
B: 0.2 M NaOH
50 mL of A + x mL of B, diluted to a total of 200 mL
x
3.7
7.5
12.2
17.7
23.9

pH
4.2
4.4
4.6
4.8
5.0

x
30.0
35.5
39.8
43.0
45.5

pH
5.2
5.4
5.6
5.8
6.0

10. Maleate Buffer.
Stocks solutions
A: 0.2 M solution of acid sodium maleate (8 g of NaOH + 23.2 g of maleic acid or
19.6 g of maleic anhydride in 1 L)
B: 0.2 M NaOH
50 mL of A + x mL of B, diluted to a total of 200 mL
x
7.2
10.5
15.3
20.8
26.9

pH
5.2
5.4
5.6
5.8
6.0

x
33.0
38.0
41.6
44.4

pH
6.2
6.4
6.6
6.8

11. Cacodylate Buffer.
Stocks solutions
A: 0.2 M solution of sodium cacodylate (42.8 g of Na(CH3)2AsO2.3H2O in 1 L)
B: 0.2 M HCl
50 mL of A + x mL of B, diluted to a total of 200 mL
x
2.7
4.2
6.3
9.3
13.3
18.3
23.8

jun 14, 2004

pH
7.4
7.2
7.0
6.8
6.6
6.4
6.2

x
29.6
34.8
39.2
43.0
45.0
47.0

pH
6.0
5.8
5.6
5.4
5.2
5.0

Pag.5

12. Phosphate Buffer.
Stocks solutions
A: 0.2 M solution of monobasic sodium phospate (27.8 g in 1 L)
B: 0.2 M solution of dibasic sodium phosphate (53.65 g of Na2HPO4.7H2O or 71.7 g
of Na2HPO4.12H2O in 1 L).
x mL of A + y mL of B, diluted to a total of 200 mL
x
93.5
92.0
90.0
87.7
85.0
81.5
77.5
73.5
68.5
62.5
56.5
51.0

y
6.5
8.0
10.0
12.3
15.0
18.5
22.5
26.5
31.5...

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