常用缓冲溶液的配制2

/常用缓冲溶液的配制方法1．甘氨酸–盐酸缓冲液（0.05mol/L）X毫升0.2mol/L甘氨酸+Y毫升0.2mol/LHCI，再加水稀释至200毫升pHXYpHXY2.02.42.62.85050505044.032.424.216.83.03.23.43.65050505011.48.26.45.0甘氨酸分子量=75.07，0.2mol/L甘氨酸溶液含15.01克/

2．邻苯二甲酸–盐酸缓冲液（0.05mol/L）X毫升0.2mol/L邻苯二甲酸氢钾+0.2mol/LHCl，再加水稀释到20毫升pH(20℃XYpH(20℃XY2.22.42.62.83.0555554.0703.9603.2952.6422.0223.23.43.63.855551.4700.9900.5970.263邻苯二甲酸氢钾分子量=204.23，0.2mol/L邻苯二甲酸氢溶液含40.85克/

3．磷酸氢二钠–柠檬酸缓冲液pH0.2mol/LNa2HPO4（毫升）0.1mol/L柠檬酸（毫升）pH0.2mol/LNa2HPO4（毫升）0.1mol/L柠檬酸（毫升）2.22.42.62.83.03.23.43.63.84.04.24.44.64.85.00.401.242.183.174.114.945.706.447.107.718.288.829.359.8610.3010.6018.7617.8216.8315.8915.0614.3013.5612.9012.2911.7211.1810.6510.149.705.25.45.65.86.06.26.46.66.87.07.27.47.67.88.010.7211.1511.6012.0912.6313.2213.8514.5515.4516.4717.3918.1718.7319.1519.459.288.858.407.917.376.786.155.454.553.532.611.831.270.850.55Na2HPO4分子量=14.98，0.2mol/L溶液为28.40克/Na2HPO4·2H2O分子量=178.05，0.2mol/L溶液含35.01克/C4H2O7·H2O分子量=210.14，0.1mol/L溶液为21.01克/

4．柠檬酸–氢氧化钠-盐酸缓冲液pH钠离子浓度（mol/L）柠檬酸（克）C6H8O7·H2O氢氧化钠(克)NaOH97%盐酸（毫升）HCl（浓）最终体积（升）①2.23.13.34.35.35.86.50.200.200.200.200.350.450.3821021021021024528526684838383144186156160116106456810512610101010101010①

使用时可以每升中加入1克克酚，若最后pH值有变化，再用少量50%

5．柠檬酸–柠檬酸钠缓冲液（0.1mol/L）pH0.1

mol/L柠檬酸（毫升）0.1

mol/L柠檬酸钠（毫升）pH0.1

mol/L柠檬酸（毫升）0.1

mol/L柠檬酸钠（毫升）3.03.23.43.63.84.04.24.44.64.818.617.216.014.914.013.112.311.410.39.21.42.84.05.16.06.97.78.69.710.85.05.25.45.65.86.06.26.46.68.27.36.45.54.73.82.82.01.411.812.713.614.515.316.217.218.018.6柠檬酸C6H8O7·H2O：分子量210.14，0.1mol/L溶液为21.01克/柠檬酸钠Na3C6H5O7·2H2O：分子量294.12，0.1mol/L溶液为29.41克

6．乙酸–乙酸钠缓冲液（0.2mol/L）

pH(18℃0.2

mol/LNaAc(毫升)0.3

mol/LHAc(毫升)

pH(18℃0.2

mol/LNaAc(毫升)0.3

mol/LHAc(毫升)2.63.84.04.24.44.60.751.201.802.653.704.909.258.808.207.356.305.104.85.05.25.45.65.85.907.007.908.609.109.404.103.002.101.400.900.60Na2Ac·3H2O分子量=136.09，0.2mol/L溶液为27.22克7．磷酸盐缓冲液（1）磷酸氢二钠–磷酸二氢钠缓冲液（0.2）

pH0.2

mol/LNa2HPO4(毫升)0.3

mol/LNaH2PO4(毫升)

pH0.2

mol/LNa2HPO4(毫升)0.3

mol/LNaH2PO4(毫升)5.85.96.06.16.26.36.46.56.66.76.86.98.010.012.315.018.522.526.531.537.543.549.555.092.090.087.785.081.577.573.568.562.556.551.045.07.07.17.27.37.47.57.67.77.87.98.061.067.072.077.081.084.087.089.591.593.0

94.739.033.028.023.019.016.013.010.58.57.05.3Na2HPO4·2H2O分子量=178.05，0.2mol/L溶液为85.61克/Na2HPO4·12H2O分子量=358.22，0.2mol/L溶液为71.64克/Na2HPO4·2H2O分子量=156.03，0.2mol/L溶液为31.21克/（2）磷酸氢二钠–磷酸二氢钾缓冲液（1/15mol/L）pHmol/L/15Na2HPO4(毫升)mol/L//15KH2PO4(毫升)pHmol/L//15Na2HPO4(毫升)mol/L//15KH2PO4(毫升)4.925.295.916.246.476.646.816.980.100.501.002.003.004.005.006.009.909.509.008.007.006.005.004.007.177.387.738.048.348.678.187.008.009.009.509.759.9010.003.002.001.000.500.250.100Na2HPO4·2H2O分子量=178.05，1/15M溶液为11.876克/KH2PO4分子量=136.09，1/15M溶液为9.078克/

8．磷酸二氢钾–氢氧化钠缓冲液（0.05M）X毫升0.2MK2PO4+Y毫升0.2NNaOH加水稀释至29毫升pH(20℃X(毫升)Y（毫升）pH(20℃X(毫升)Y（毫升）5.86.06.26.46.66.85555550.3720.5700.8601.2601.7802.3657.07.27.47.67.88.05555552.9633.5003.9504.2804.5204.680

9．巴比妥钠-盐酸缓冲液（18℃）pH0.04M巴比妥钠溶液0.2V盐酸pH0.04M巴比妥钠溶液(毫升)0.2N盐酸(毫升)6.87.07.27.47.67.88.08.2100

10010010010010010010018.417.816.7

15.313.411.479.397.218.48.68.89.09.29.49.61001001001001001001001005.213.822.521.651.130.700.35巴比妥钠盐分子量=206.18;0.04M溶液为8.25克/

10．Tris–盐酸缓冲液（0.05M，25℃50毫升0.1M三羟甲基氨基甲烷（Tris）溶液与X毫升0.1N盐酸混匀后，加水稀释至100毫升。pHX(毫升)pHX(毫升)7.107.207.307.407.507.607.707.807.908.0045.744.743.442.040.338.536.634.532.0

29.28.108.208.308.408.508.608.708.808.9026.222.9

19.917.214.712.410.38.57.0三羟甲基氨基甲烷（Tris）HOCH2CH2OHCHOCH2NH2分子量=121.14;0.1M溶液为12.114克/升。Tris

11．硼酸–硼砂缓冲液（0.2M硼酸根）pH0.05M硼砂（毫升）0.2M硼砂（毫升）pH0.05M硼砂（毫升）0.2M硼酸（毫升）7.47.67.88.01.0

1.52.03.09.08.58.07.08.28.48.79.03.54.56.08.06.55.54.02.0硼砂Na2B4O7·H2O,分子量=381.43;0.05M溶液（=0.2M硼酸根）含19.07克/硼酸H2BO3,分子量=61.84,0.2M溶液为12.37克/硼砂易失去结晶水，必须在带塞的瓶中保存。

11．硼酸-硼砂缓冲液（0.2M硼酸根）pH0.05M硼砂（毫升）0.2M硼酸（毫升）pH0.05M硼砂（毫升）0.2M硼酸（毫升）7.47.67.88.01.01.52.03.09.0

8.58.07.08.28.48.79.03.54.56.08.06.55.54.0

2.0硼砂Na2B4O7·10H2O,分子量=381.43;0.05M溶液（=0.2M硼酸根）含19.07克/硼酸H2BO3，分子量=61.84,0.2M溶液为12.37克/硼砂 易失去结晶水，必须在带塞的瓶中保存。

12．甘氨酸–氢氧化钠缓冲液（0.05M）X毫升0.2M甘氨酸+Y毫升0.2NnaOH加水稀释至200毫升pHXYpHXY8.68.89.09.29.450505050504.06.08.812.016.89.69.810.010.410.6505050505022.427.232.038.645.5甘氨酸分子量=75.07;0.2M溶液含15.01克/

13．硼砂-氢氧化钠缓冲液（0.05M硼酸根）X毫升0.05M硼砂+Y毫升0.2NNaOH加水稀释至200毫升pHXYpHXY9.39.49.65050506.0

11.023.09.810.010.150505034.043.046.0硼砂Na2B4O7·10H2O,分子量=381.43;0.05M溶液为19.07克/

14．碳酸钠-碳酸氢钠缓冲液（0.1M）Ca2+、Mg2+存在时不得使用pH0.1MNa2CO3(毫升)0.1MN2HCO3(毫升)20379.169.409.519.789.9010.1410.2810.5310.838.779.129.409.509.729.9010.0810.2810.57123456789987654321Na2CO2·10H2O分子量=286.2;0.1M溶液为28.62克/N2HCO3分子量=84.0;0.1M溶液为8.40克/

15．“PBS”缓冲液PH7.67.47.27.0H2ONaClNa2HPO4NaH2PO410008.52.20.110008.52.20.210008.52.20.3100ml8.5g2.2g0.4g

附录四常用缓冲溶液的配制

（一）磷酸氢二钠-柠檬酸缓冲液pH0.2mol/LNa2HPO4(mL)0.1mol/L柠檬酸(mL)pH0.2mol/LNa2HPO4(mL)0.1

mol/L柠檬酸(mL)2.22.42.62.83.03.23.43.63.84.04.24.44.64.85.00.401.242.183.174.114.945.706.447.107.718.288.829.359.8610.3019.6018.7617.8216.8315.8915.0614.3013.5612.9012.2911.7211.1810.6510.149.705.25.45.65.86.06.26.46.66.87.07.27.47.67.88.010.7211.1511.6012.0912.6313.2213.8514.5515.4516.4717.3918.1718.7319.1519.459.288.858.407.917.376.786.155.454.553.532.611.831.270.850.55Na2HPO4分子量＝141.98；0.2mol/L溶液为28.40g/L。Na2HPO4·2H2O分子量＝178.05；0.2mol/L溶液为35.61g/L。Na2HPO4·12H2O分子量＝358.22；0.2mol/L溶液为71.64g/L。C6H8O7·H2O分子量＝210.14；0.1mol/L溶液为21.01g/L。

（二）柠檬酸-柠檬酸钠缓冲液（0.1mol/L）pH0.1

mol/L柠檬酸(mL)0.1mol/L柠檬酸钠(mL)pH0.1mol/L柠檬酸(mL)0.1mol/L柠檬酸钠(mL)3.018.61.45.08.211.83.217.22.85.27.312.73.416.04.05.46.413.63.614.95.15.65.514.53.814.06.05.84.715.34.013.16.96.03.816.24.212.37.76.22.817.24.411.48.66.42.018.04.610.39.76.61.418.64.89.210.8

柠檬酸：C6H8O7·H2O分子量＝210.14；0.1mol/L溶液为21.01g/L。柠檬酸钠：Na3C6H5O7·2H2O分子量＝294.12；0.1mol/L溶液为29.41g/L。

（三）醋酸-醋酸钠缓冲液（0.2mol/L）pH(18℃0.2mol/LNaAc

(mL)0.2mol/LHAc

(mL)pH(18℃0.2mol/LNaAc

(mL)0.2mol/LHAc

(mL)3.60.759.354.85.904.103.81.208.805.07.003.004.01.808.205.27.902.104.22.657.355.48.601.404.43.706.305.69.100.904.64.905.105.86.400.60NaAc·3H2O分子量＝136.09；0.2mol/L溶液为27.22g/L。冰乙酸11.8mL稀释至1L（需标定）。

（四）磷酸二氢钾-氢氧化钠缓冲液（0.05mol/L）X毫升0.2mol/LKH2PO4+Y毫升0.2mol/LNaOH加水稀释至20毫升。pH

(20℃X

(mL)Y

(mL)pH

(20℃

X

(mL)

Y

(mL)5.850.3727.052.9636.050.5707.253.5006.250.8607.4

3.9506.451.2607.654.2806.651.7807.854.5206.852.3658.054.680

（五）磷酸氢二钠-磷酸二氢钠缓冲液（0.2mol/L）pH0.2mol/LNa2HPO4

(mL)0.2mol/LNaH2PO4(mL)pH0.2mol/LNa2HPO4

(mL)0.2mol/LNaH2PO4(mL)5.88.092.07.061.039.05.910.090.07.167.033.06.012.387.77.272.028.06.115.085.07.377.023.06.218.581.57.481.019.06.322.577.57.584.016.06.426.573.57.687.013.06.531.568.57.789.510.56.637.562.57.891.58.56.743.556.57.993.07.06.849.051.08.094.75.36.955.045.0

Na2HPO4·2H2O分子量＝178.05；0.2mol/L溶液为35.61g/L。Na2HPO4·12H2O分子量＝358.22；0.2mol/L溶液为71.64g/L。NaH2PO4·H2O分子量＝138.01；0.2mol/L溶液为27.6g/L。NaH2PO4·2H2O分子量＝156.03；0.2mol/L溶液为31.21g/L。（六）巴比妥纳-盐酸缓冲液pH（18℃0.04mol/L巴比妥钠(mL)0.2mol/LHCl(mL)PH（18℃0.04mol/L巴比妥钠(mL)0.2mol/LHCl(mL)6.810018.48.41005.217.010017.88.61003.827.210016.78.81002.527.410015.39.01001.657.610013.49.21001.137.810011.479.41000.708.01009.399.61000.358.21007.21

巴比妥钠分子量＝206.18；0.04mol/L溶液为8.25g/L。

（七）Tris-HCl缓冲液（0.05mol/L）50毫升0.1mol/L三羟甲基氨基甲烷（Tris）溶液与X毫升0.1mol/L盐酸混匀并稀释至100毫升。pH

(25℃X

(mL)pH

(25℃X

(mL)7.1045.78.1026.27.2044.78.2022.97.3043.48.3019.97.4042.08.4017.27.5040.38.5014.77.6038.58.6012.47.7036.68.7010.37.8034.58.808.57.9032.08.907.08.0029.2

Tris分子量＝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，使用时注意将瓶盖严。（八）、硼酸-硼砂缓冲液(0.2mol/L硼酸根)pH0.05mol/L硼砂(mL)0.2mol/L硼酸(mL)pH0.05mol/L硼砂(mL)0.2mol/L硼酸(mL)7.41.09.08.23.56.57.61.58.58.44.55.57.82.08.08.76.04.08.03.07.09.08.02.0硼砂：Na2B4O7·10H2O分子量＝381.43；0.05mol/L溶液（等于0.2mol/L硼酸根）含19.07g/L。硼酸：H3BO3分子量＝61.84；0.2mol/L的溶液为12.37g/L。硼砂易失去结晶水，必须在带塞的瓶中保存。

（九）硼砂-氢氧化纳缓冲液（0.05mol/L硼酸根）X