常用缓冲溶液配制方法

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/升。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/升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/升。Na2HPO4·2H2O分子量=178.05，0.2mol/L溶液含35.01克/升。C4H2O7·H2O分子量=210.14，0.1mol/L溶液为21.01克/升。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%氢氧化钠溶液或浓盐酸调节，冰箱保存。5．柠檬酸–柠檬酸钠缓冲液（0.1mol/L）pH0.1mol/L柠檬酸（毫升）0.1mol/L柠檬酸钠（毫升）pH0.1mol/L柠檬酸（毫升）0.1mol/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/升。柠檬酸钠Na3C6H5O7·2H2O：分子量294.12，0.1mol/L溶液为29.41克/毫升。6．乙酸–乙酸钠缓冲液（0.2mol/L）pH(18℃)0.2mol/LNaAc(毫升)0.3mol/LHAc(毫升)pH(18℃)0.2mol/LNaAc(毫升)0.3mol/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/升。7．磷酸盐缓冲液（1）磷酸氢二钠–磷酸二氢钠缓冲液（0.2）pH0.2mol/LNa2HPO4(毫升)0.2mol/LNaH2PO4(毫升)pH0.2mol/LNa2HPO4(毫升)0.2mol/LNaH2PO4(毫升)5.85.96.06.16.26.36.46.56.66.76.86.98.010.012.315.018.522.526.531.537.543.549.555.092.090.087.785.081.577.573.568.562.556.551.045.07.07.17.27.37.47.57.67.77.87.98.061.067.072.077.081.084.087.089.591.593.094.739.033.028.023.019.016.013.010.58.57.05.3Na2HPO4·2H2O分子量=178.05，0.2mol/L溶液为85.61克/升。Na2HPO4·12H2O分子量=358.14，0.2mol/L溶液为71.628克/升。NaH2PO4·2H2O分子量=156.01，0.2mol/L溶液为31.202克/升。磷酸盐是生物化学研究中使用最广泛的一种缓冲剂，由于它们是二级解离，有二个pKa值，所以用它们配制的缓冲液，pH范围最宽：NaH2PO4：pKa1＝2.12，pKa2＝7.21；Na2HPO4：pKa1＝7.21，pKa2＝12.32配酸性缓冲液：用NaH2PO4，pH＝1～4，配中性缓冲液：用混合的两种磷酸盐，pH＝6～8，配碱性缓冲液：用Na2HPO4，pH＝10～12。用钾盐比钠盐好，因为低温时钠盐难溶，钾盐易溶，但若配制SDS-聚丙烯酰胺凝胶电泳的缓冲液时，只能用磷酸钠而不能用磷酸钾，因为SDS(十二烷基硫酸钠)会与钾盐生成难溶的十二烷基硫酸钾。磷酸盐缓冲液的优点为：①容易配制成各种浓度的缓冲液；②适用的pH范围宽；③pH受温度的影响小；④缓冲液稀释后pH变化小，如稀释10倍后pH的变化小于0.1。其缺点为：①易与常见的钙Ca2+离子、镁Mg2+离子以及重金属离子缔合生成沉淀；②会抑制某些生物化学过程，如对某些酶的催化作用会产生某种程度的抑制作用。（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)4.925.295.910.100.501.009.909.509.007.177.387.737.008.009.003.002.001.006.246.476.646.816.982.003.004.005.006.008.007.006.005.004.008.048.348.678.189.509.759.9010.000.500.250.100Na2HPO4·2H2O分子量=178.05，1/15M溶液为11.876克/升。KH2PO4分子量=136.09，1/15M溶液为9.078克/升。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）5.86.06.26.46.66.85555550.3720.5700.8601.2601.7802.3657.07.27.47.67.88.05555552.9633.5003.9504.2804.5204.6809．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)6.87.07.27.47.67.88.08.210010010010010010010010018.417.816.715.313.411.479.397.218.48.68.89.09.29.49.61001001001001001001001005.213.822.521.651.130.700.35巴比妥钠盐分子量=206.18;0.04M溶液为8.25克/升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)7.107.207.307.407.507.607.707.8045.744.743.442.040.338.536.634.58.108.208.308.408.508.608.708.8026.222.919.917.214.712.410.38.57.908.0032.029.28.907.0三羟甲基氨基甲烷（Tris）HOCH2CH2OHCHOCH2NH2分子量=121.14;0.1M溶液为12.114克/升。Tris溶液可从空气中吸收二氧化碳，使用时注意将瓶盖严。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）7.47.67.88.01.01.52.03.09.08.58.07.08.28.48.79.03.54.56.08.06.55.54.02.0硼砂Na2B4O7·10H2O,分子量=381.37;0.05M溶液（=0.2M硼酸根）含19.07克/升。硼酸H3BO3,分子量=61.83,0.2M溶液为12.37克/升。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/升。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pHXYpHXY9.39.49.65050506.011.023.09.810.010.150505034.043.046.0硼砂Na2B4O7·10H2O,分子量=381.43;0.05M溶液为19.07克/升。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)20℃37℃9.169.409.519.788.779.129.409.50123498769.9010.1410.2810.5310.839.729.9010.0810.2810.575678954321Na2CO2·10H2O分子量=286.2;0.1M溶液为28.62克/升。N2HCO3分子量=84.0;0.1M溶液为8.40克/升。15．“PBS”缓冲液pH7.67.47.27.0H2ONaClNa2HPO4NaH2PO410008.52.20.110008.52.20.210008.52.20.3100ml8.5g2.2g0.4gpH标准缓冲溶液StandardpHBufferSolutions名称(Name)配制(Compoundingway)不同温度时的pH值(pHindifferenttemperatures)0℃5℃10℃15℃20℃25℃30℃35℃40℃草酸盐标准缓冲溶液c[KH3(C2O4)2·2H2O]为0.05mol/L。称取1