缓冲液配制方法.docx
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缓冲液配制方法.docx
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缓冲液配制方法
一.常用缓冲溶液的配制方法
1.甘氨酸—盐酸缓冲液(0.05mol/L)
xml0.2mol/L甘氨酸+yml0.2mol/LHCl,再加水稀释至200ml
pH
x/ml
y/ml
pH
x/ml
y/ml
2.2
50
44.0
3.0
50
11.4
2.4
50
32.4
3.2
50
8.2
2.6
50
24.2
3.4
50
6.4
2.8
50
16.8
3.6
50
5.0
甘氨酸Mr=75.07,0.2mol/L甘氨酸溶液含15.01g/L
2.甘氨酸—氢氧化钠缓冲液(0.05mol/L)
xml0.2mol/L甘氨酸+yml0.2mol/LNaOH,加水稀释至200ml
pH
x/ml
y/ml
pH
x/ml
y/ml
8.6
50
4.0
9.6
50
22.4
8.8
50
6.0
9.8
50
27.2
9.0
50
8.8
10.0
50
32.0
9.2
50
12.0
10.4
50
38.6
9.4
50
16.8
10.6
50
45.5
甘氨酸Mr=75.07,0.2mol/L甘氨酸溶液为15.01g/L
3.邻苯二甲酸—盐酸缓冲液(0.05mol/L,20℃)
xml0.2mol/L邻苯二甲酸氢钾+yml0.2mol/LHCl,再加水稀释到20ml
pH
x/ml
y/ml
pH
x/ml
y/ml
2.2
5
4.670
3.2
5
1.470
2.4
5
3.960
3.4
5
0.990
2.6
5
3.295
3.6
5
0.597
2.8
5
2.642
3.8
5
0.263
3.0
5
2.032
邻苯二甲酸氢钾Mr=204.23,0.2mol/L邻苯二甲酸氢钾溶液含40.85g/L
4.柠檬酸—氢氧化钠—盐酸缓冲液
pH
Na+浓度
/mol/L
柠檬酸/g
C6H8O7·H2O
氢氧化钠/g
NaOH(97%)
盐酸/ml
HCl(浓)
最终体积/L*
2.2
0.20
210
84
160
10
3.1
0.20
210
83
116
10
3.3
0.20
210
83
106
10
4.3
0.20
210
83
45
10
5.3
0.35
245
144
68
10
5.8
0.45
285
186
105
10
6.5
0.38
266
156
126
10
使用时可以每升中加入1g酚,若最后pH值有变化,再用少量50%氢氧化钠溶液或浓盐酸调节,置冰箱保存。
5.磷酸氢二钠—柠檬酸缓冲液
pH
0.2ol/LNa2HPO4/ml
0.1mol/L
柠檬酸/ml
pH
0.2mol/LNa2HPO4/ml
0.1mol/L
柠檬酸/ml
2.2
0.40
19.60
5.2
10.72
9.28
2.4
1.24
18.76
5.4
11.15
8.85
2.6
2.18
17.82
5.6
11.60
8.40
2.8
3.17
16.83
5.8
12.09
7.91
3.0
4.11
15.89
6.0
12.63
7.37
3.2
4.94
15.06
6.2
13.22
6.78
3.4
5.70
14.30
6.4
13.85
6.15
3.6
6.44
13.56
6.6
14.55
5.45
3.8
7.10
12.90
6.8
15.45
4.55
4.0
7.71
12.29
7.0
16.47
3.53
4.2
8.28
11.72
7.2
17.39
2.61
4.4
8.82
11.18
7.4
18.17
1.83
4.6
9.35
10.65
7.6
18.73
1.27
4.8
9.86
10.14
7.8
19.15
0.85
5.0
10.30
9.70
8.0
19.45
0.55
Na2HPO4Mr=141.98,0.2mol/L溶液为28.40g/L,
Na2HPO4·2H2OMr=178.05,0.2mol/L溶液为35.61g/L
C6H8O7·H2OMr=210.14,0.1mol/L溶液为21.01g/L
6.柠檬酸—柠檬酸钠缓冲液(0.1mol/L)
pH
0.1mol/L
柠檬酸/ml
0.1mol/L
柠檬酸钠/ml
pH
0.1mol/L
柠檬酸/ml
0.1mol/L
柠檬酸钠/ml
3.0
18.6
1.4
5.0
8.2
11.8
3.2
17.2
2.8
5.2
7.3
12.7
3.4
16.0
4.0
5.4
6.4
13.6
3.6
14.9
5.1
5.6
5.5
14.5
3.8
14.0
6.0
5.8
4.7
15.3
4.0
13.1
6.9
6.0
3.8
16.2
4.2
12.3
7.7
6.2
2.8
17.2
4.4
11.4
8.6
6.4
2.0
18.0
4.6
10.3
9.7
6.6
1.4
18.6
4.8
9.2
10.8
柠檬酸C6H8O7·H2O,Mr=210.14,0.1mol/L溶液为21.01g/L
柠檬酸钠Na3C6H5O7·2H2O,Mr=294.12,0.1mol/L溶液为29.41g/L
7.乙酸—乙酸钠缓冲液(0.2mol/L,18℃)
pH
0.2mol/L
NaAc/ml
0.2mol/L
HAc/ml
pH
0.2mol/L
NaAc/ml
0.2mol/L
HAc/ml
3.6
0.75
9.25
4.8
5.90
4.10
3.8
1.20
8.80
5.0
7.00
3.00
4.0
1.80
8.20
5.2
7.90
2.10
4.2
2.65
7.35
5.4
8.60
1.40
4.4
3.70
6.30
5.6
9.10
0.90
4.6
4.90
5.10
5.8
9.40
0.60
NaAc·3H2O,Mr=136.09,0.2mol/L溶液为27.22g/L
8.邻苯二甲酸氢钾—氢氧化钠缓冲液
50ml0.1mol/L邻苯二甲酸氢钾+xml0.1mol/L氢氧化钠,加水稀释到100ml
pH
x/ml
pH
x/ml
pH
x/ml
4.1
1.3
4.8
16.5
5.5
36.6
4.2
3.0
4.9
19.4
5.6
38.8
4.3
4.7
5.0
22.6
5.7
40.6
4.4
6.6
5.1
25.5
5.8
42.3
4.5
8.7
5.2
28.8
5.9
43.7
4.6
11.1
5.3
31.6
4.7
13.6
5.4
34.1
邻苯二甲酸氢钾Mr=204.23,0.1mol/L溶液为20.42g/L
9.磷酸盐缓冲液
(1)磷酸氢二钠—磷酸二氢钠缓冲液(0.2mol/L)
pH
0.2mol/L
Na2HPO4/ml
0.2mol/L
NaH2PO4/ml
pH
0.2mol/L
Na2HPO4/ml
0.2mol/L
NaH2PO4/ml
5.8
8.0
92.0
7.0
61.0
39.0
5.9
10.0
90.0
7.1
67.0
33.0
6.0
12.3
87.7
7.2
72.0
28.0
6.1
15.0
85.0
7.3
77.0
23.0
6.2
18.5
81.5
7.4
81.0
19.0
6.3
22.5
77.5
7.5
84.0
16.0
6.4
26.5
73.5
7.6
87.0
13.0
6.5
31.5
68.5
7.7
89.5
10.5
6.6
37.5
62.5
7.8
91.5
8.5
6.7
43.5
56.5
7.9
93.0
7.0
6.8
49.0
51.0
8.0
94.7
5.3
6.9
55.0
45.0
Na2HPO4·2H2O,Mr=178.05,0.2mol/L溶液为35.61g/L
Na2HPO4·12H2O,Mr=358.22,0.2mol/L溶液为71.64g/L
NaH2PO4·H2O,Mr=138.01,0.2mol/L溶液为27.6g/L
NaH2PO4·2H2O,Mr=156.03,0.2mol/L溶液为31.21g/L
(2)磷酸氢二钠—磷酸二氢钾缓冲液(1/15mol/L)
pH
1/15mol/L
Na2HPO4/ml
1/15mol/L
KH2PO4/ml
pH
1/15mol/L
Na2HPO4/ml
1/15mol/L
KH2PO4/ml
4.92
0.10
9.90
7.17
7.00
3.00
5.29
0.50
9.50
7.38
8.00
2.00
5.91
1.00
9.00
7.73
9.00
1.00
6.24
2.00
8.00
8.04
9.50
0.50
6.47
3.00
7.00
8.34
9.75
0.25
6.64
4.00
6.00
8.67
9.90
0.10
6.81
5.00
5.00
8.18
10.00
0
6.98
6.00
4.00
Na2HPO4·2H2O,Mr=178.05,1/15mol/L溶液为11.876g/L
KH2PO4,Mr=136.09,1/15mol/L溶液为9.078g/L
10.磷酸氢二钠—氢氧化钠缓冲液
50ml0.05mol/L磷酸氢二钠+xml0.1mol/LNaOH,加水稀释至100ml
pH
x/ml
pH
x/ml
pH
x/ml
10.9
3.3
11.3
7.6
11.7
16.2
11.0
4.1
11.4
9.1
11.8
19.4
11.1
5.1
11.5
11.1
11.9
23.0
11.2
6.3
11.6
13.5
12.0
26.9
Na2HPO4·2H2O,Mr=178.05,0.05mol/L溶液为8.90g/L
Na2HPO4·12H2O,Mr=358.22,0.05mol/L溶液为17.91g/L
11.磷酸二氢钾—氢氧化钠缓冲液(0.05mol/L,20℃)
xml0.2mol/LKH2PO4+yml0.2mol/LNaOH,加水稀释至20ml
pH
x/ml
y/ml
pH
x/ml
y/ml
5.8
5
0.372
7.0
5
2.963
6.0
5
0.570
7.2
5
3.500
6.2
5
0.860
7.4
5
3.950
6.4
5
1.260
7.6
5
4.280
6.6
5
1.780
7.8
5
4.520
6.8
5
2.365
8.0
5
4.680
12.硼砂—盐酸缓冲液(0.05mol/L硼酸根)
50ml0.025mol/L硼砂+xml0.1mol/L盐酸,加水稀释至100ml
pH
x/ml
pH
x/ml
pH
x/ml
8.00
20.5
8.4
16.6
8.8
9.4
8.10
19.7
8.5
15.2
8.9
7.1
8.20
18.8
8.6
13.5
9.0
4.6
8.30
17.7
8.7
11.6
9.1
2.0
硼砂Na2B4O7·10H2O,Mr=381.43,0.025mol/L溶液为9.53g/L
13.巴比妥钠—盐酸缓冲液(18℃)
pH
0.04mol/L
巴比妥钠溶液/ml
0.2mol/L
盐酸/ml
pH
0.04mol/L
巴比妥钠溶液/ml
0.2mol/L
盐酸/ml
6.8
100
18.4
8.4
100
5.21
7.0
100
17.8
8.6
100
3.82
7.2
100
16.7
8.8
100
2.52
7.4
100
15.3
9.0
100
1.65
7.6
100
13.4
9.2
100
1.13
7.8
100
11.47
9.4
100
0.70
8.0
100
9.39
9.6
100
0.35
8.2
100
7.21
巴比妥钠盐Mr=206.18,0.04mol/L溶液为8.25g/L
14.Tris—盐酸缓冲液(0.05mol/L,25℃)
50ml0.1mol/L三羟甲基氨基甲烷(Tris)溶液与xml0.1mol/L盐酸混匀后,加水稀释至100ml
pH
x/ml
pH
x/ml
7.10
45.7
8.10
26.2
7.20
44.7
8.20
22.9
7.30
43.4
8.30
19.9
7.40
42.0
8.40
17.2
7.50
40.3
8.50
14.7
7.60
38.5
8.60
12.4
7.70
36.6
8.70
10.3
7.80
34.5
8.80
8.5
7.90
32.0
8.90
7.0
8.00
29.2
9.00
5.7
注:
三羟甲基氨基甲烷(Tris)
Mr=121.14,0.1mol/L溶液为12.114g/L。
Tris溶液可从空气中吸收二氧化碳,保存时注意密封。
15.硼酸—硼砂缓冲液(0.2mol/L硼酸根)
pH
0.05mol/L
硼砂/ml
0.2mol/L
硼酸/ml
pH
0.05mol/L
硼砂/ml
0.2mol/L
硼酸/ml
7.4
1.0
9.0
8.2
3.5
6.5
7.6
1.5
8.5
8.4
4.5
5.5
7.8
2.0
8.0
8.7
6.0
4.0
8.0
3.0
7.0
9.0
8.0
2.0
硼砂Na2B4O7·10H2O,Mr=381.43,0.05mol/L溶液(=0.2mol/L硼酸根)为19.07g/L,硼砂易失去结晶水,必须密闭保存
硼酸H3BO3,Mr=61.84,0.2mol/L溶液为12.37g/L
16.硼砂—氢氧化钠缓冲液(0.05mol/L硼酸根)
xml0.05mol/L硼砂+yml0.2mol/LNaOH,加水稀释至200ml
pH
x/ml
y/ml
pH
x/ml
y/ml
9.3
50
6.0
9.8
50
34.0
9.4
50
11.0
10.0
50
43.0
9.6
50
23.0
10.1
50
46.0
硼砂Na2B4O7·10H2O,Mr=381.43,0.05mol/L溶液为19.07g/L
17.碳酸钠—碳酸氢钠缓冲液(0.1mol/L)
Ca2+、Mg2+存在时不得使用
pH
0.1mol/L碳酸钠/ml
0.1mol/L碳酸氢钠/ml
20℃
37℃
9.16
8.77
1
9
9.40
9.12
2
8
9.51
9.40
3
7
9.78
9.50
4
6
9.90
9.72
5
5
10.14
9.90
6
4
10.28
10.08
7
3
10.53
10.28
8
2
10.83
10.57
9
1
无水碳酸钠Mr=105.99,0.1mol/L溶液为10.60g/L
碳酸氢钠Mr=84.01,0.1mol/L溶液为8.40g/L
18.碳酸氢钠—氢氧化钠缓冲液(0.025mol/L碳酸氢钠)
50ml0.05mol/L碳酸氢钠+xml0.1mol/LNaOH,加水稀释至100ml
pH
x/ml
pH
x/ml
pH
x/ml
9.6
5.0
10.1
12.2
10.6
19.1
9.7
6.2
10.2
13.8
10.7
20.2
9.8
7.6
10.3
15.2
10.8
21.2
9.9
9.1
10.4
16.5
10.9
22.0
10.0
10.7
10.5
17.8
11.0
22.7
碳酸氢钠Mr=84.01;0.05mol/L溶液为4.20g/L
19.氯化钾—氢氧化钠缓冲液
25ml0.2mol/LKCl+xml0.2mol/LNaOH,加水稀释至100ml
pH
x/ml
pH
x/ml
pH
x/ml
12.0
6.0
12.4
16.2
12.8
41.2
12.1
8.0
12.5
20.4
12.9
53.0
12.2
10.2
12.6
25.6
13.0
66.0
12.3
12.8
12.7
32.2
KClMr=74.55;0.2mol/L溶液为14.91g/L
二.常用市售酸碱的浓度
溶质
分子式
Mr
mol/L
g/L
重量百分比
比重
配制1mol/L溶液的加入量/ml/L
冰乙酸
CH3COOH
60.05
17.4
1045
99.5
1.05
57.5
乙酸
60.05
6.27
376
36
1.045
159.5
甲酸
HCOOH
46.02
23.4
1080
90
1.20
42.7
盐酸
HCl
36.5
11.6
2.9
424
105
36
10
1.18
1.05
86.2
344.8
硝酸
HNO3
63.02
15.99
14.9
13.3
1008
938
837
71
67
61
1.42
1.40
1.37
62.5
67.1
75.2
高氯酸
HClO4
100.5
11.65
9.2
1172
923
70
60
1.67
1.54
85.8
108.7
磷酸
H3PO4
80.0
18.1
1445
85
1.70
55.2
硫酸
H2SO4
98.1
18.0
1766
96
1.84
55.6
氢氧化铵
NH4OH
35.0
14.8
251
28
0.898
67.6
氢氧化钾
KOH
56.1
13.5
1.94
757
109
50
10
1.52
1.09
74.1
515.5
氢氧化钠
NaOH
40.0
19.1
2.75
763
111
50
10
1.53
1.11
52.4
363.6
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