常用缓冲液配方及缓冲范围

常见缓冲液的缓冲范围缓冲液名称及常用浓度缓冲pH范围主要物质分子量MES(2吗啉代乙磺酸)5.5-6.7195.2Bis-Tris5.8-7.2209.2HEPES6.8-8.2238.3PIPES6.1-7.5302.4MOPS6.5-7.9209.3Tricine7.4-8.8179.2TEA(三乙醇胺)7.4-8.3149.2甘氨酸-盐酸缓冲液（0.05mol/L）2.2-5.0甘氨酸Mr=75.07邻苯二甲酸-盐酸缓冲液（0.05mol/L）2.2-3.8邻苯二甲酸氢钾Mr=204.23磷酸氢二钠-柠檬酸缓冲液2.2-8.0磷酸氢二钠Mr=141.98柠檬酸-氢氧化钠-盐酸缓冲液2.2-6.5柠檬酸Mr=192.06柠檬酸-柠檬酸钠缓冲液（0.1mol/L）3.0-6.6柠檬酸Mr=192.06柠檬酸钠Mr=257.96乙酸-乙酸钠缓冲液（0.2mol/L）3.6-5.8乙酸钠Mr=81.76乙酸Mr=60.05邻苯二甲酸氢钾-氢氧化钠缓冲液4.1-5.9氢钾Mr=204.23磷酸氢二钠-磷酸二氢钠缓冲液（0.2mol/L）5.8-8.0磷酸氢二钠-磷酸二氢钾缓冲液（1/15molL）4.92-8.18磷酸二氢钾-氢氧化钠缓冲液（0.05mol/L）5.8-8.0巴比妥钠-盐酸缓冲液(18℃)6.8-9.6巴比妥钠Mr=206.18Tris-盐酸缓冲液(0.05mol/L25℃)7.10-9.00三羟甲基氨基甲烷（Tris）Mr=121.14甘氨酸-氢氧化钠缓冲液(0.05mol/L)8.6-10.6甘氨酸Mr=75.07硼砂-氢氧化钠缓冲液(0.05mol/L)9.3-10.1硼砂Na2B4O7·10H2OMr=381.43碳酸钠-碳酸氢钠缓冲液(0.1mol/L)9.16-10.83碳酸钠Mr=286.2碳酸氢钠Mr=84.0碳酸钠-氢氧化钠缓冲液(0.025mol/L)9.6-11.0磷酸氢二钠-氢氧化钠缓冲液10.9-12.0氯化钾-盐酸缓冲液（0.2mol/L）1.0-2.2氯化钾Mr=74.55氯化钾-氢氧化钠缓冲液（0.2mol/L）12.0-13.0氯化钾Mr=74.55参考下图：常见缓冲液的缓冲范围常用缓冲液的配制方法1.甘氨酸-盐酸缓冲液（0.05M）Xml0.2M甘氨酸+Yml0.2M盐酸再加水稀释至200mlpHX/mlY/mlpHX/mlY/ml2.25044.03.05011.42.45032.43.2508.22.65024.23.4506.42.85016.83.6505.0甘氨酸分子量＝75.070.2M甘氨酸溶液含15.01g/L2.邻苯二甲酸-盐酸缓冲液（0.05M）Xml0.2M邻苯二甲酸氢钾+Yml0.2M盐酸再加水稀释至200mlpHXYpHXY2.254.6703.251.4702.453.9603.450.9902.653.2953.650.5972.852.6423.850.2633.052.032邻苯二甲酸氢钾分子量＝2.4.230.2M邻苯二甲酸氢钾溶液含40.85g/L3.磷酸氢二钠-柠檬酸缓冲液pH0.2MNa2HPO4/ml0.1M柠檬酸/mlpH0.2MNa2HPO4/ml0.1M柠檬酸/ml2.20.4019.65.210.729.282.41.2418.765.411.158.852.62.1817.825.611.608.402.83.1716.835.812.097.913.04.1115.896.012.637.373.24.9415.066.213.226.783.45.7014.306.413.856.153.66.4413.566.614.555.453.87.1012.906.815.454.554.07.7112.297.016.473.534.28.2811.727.217.392.614.48.8211.187.418.171.834.69.3510.657.618.731.274.89.8610.147.819.150.855.010.309.708.019.450.55Na2HPO4分子量＝141.980.2M溶液含28.40g/LNa2HPO4·2H2O分子量＝178.050.2M溶液含35.61g/LC6H8O7·H2O分子量＝210.140.1M溶液含21.01g/L4.柠檬酸-氢氧化钠-盐酸缓冲液pH钠离子浓度/M柠檬酸C6H8O7·H2O/g氢氧化钠NaOH/g浓盐酸HCl/ml终体积/L2.20.2021084160103.10.2021083116103.30.2021083106104.30.202108345105.30.3524514468105.80.45285186105106.50.3826615612610使用时可以每升中加入1g酚，若最后pH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（0.1M）pH0.1M柠檬酸/ml0.1M柠檬酸钠/mlpH0.1M柠檬酸/ml0.1M柠檬酸钠/ml3.018.61.45.08.211.83.217.22.85.27.312.73.416.04.05.46.413.63.614.95.15.65.514.53.814.06.05.84.715.34.013.16.96.03.816.24.212.37.76.22.817.24.411.48.66.42.018.04.610.39.76.610418.64.89.210.8C6H8O7·H2O分子量＝210.140.1M溶液含21.01g/LNa3C6H5O7·2H2O分子量＝294.120.1M溶液含29.41g/L6.乙酸-乙酸钠缓冲液（0.2M）pH0.2MNaAc/ml0.2MHAC/mlpH0.2MNaAc/ml0.2MHAC/ml3.60.759.254.85.904.103.81.208.805.07.003.004.01.808.205.27.902.104.22.657.355.48.601.404.43.706.305.69.100.904.64.905.105.89.400.60NaAc分子量＝136.090.2M溶液为27.22g/L7.磷酸盐缓冲液(1)磷酸氢二钠-磷酸二氢钠缓冲液（0.2M）pH0.2MNa2HPO4/ml0.2MNaH2PO4/mlpH0.2MNa2HPO4/ml0.2MNaH2PO4/ml5.88.092.07.061.039.05.910.090.07.167.033.06.012.387.77.272.028.06.115.085.07.377.023.06.218.581.57.481.019.06.322.577.57.584.016.06.426.573.57.687.013.06.531.568.57.789.510.56.637.562.57.891.58.56.743.556.57.993.07.06.849.551.08.094.75.36.955.045.0Na2HPO4·2H2O分子量＝178.050.2M溶液含35.61g/LNa2HPO4·12H2O分子量＝358.220.2M溶液含71.64g/LNaH2PO4·H2O分子量＝138.010.2M溶液含27.6g/LNaH2PO4·2H2O分子量＝156.030.2M溶液含31.21g/L(2)磷酸氢二钠-磷酸二氢钾缓冲液（1/15M）pH1/15MNa2HPO4/ml1/15MKH2PO4/mlpH1/15MNa2HPO4/ml1/15MKH2PO4/ml4.920.109.907.177.003.005.290.509.507.388.002.005.911.009.007.739.001.006.242.008.008.049.500.506.473.007.008.349.750.256.644.006.008.679.900.106.815.005.009.1810.0006.986.004.00Na2HPO4·2H2O分子量＝178.051/15M溶液含35.61g/LKH2PO4分子量＝136.091/15M溶液含9.078g/L8.磷酸二氢钾-氢氧化钠缓冲液（0.05M）Xml0.2MKH2PO4+Yml0.2MNaOH再加水稀释至20mlpH(20℃)X/mlY/mlpH(20℃)X/mlY/ml5.850.3727.052.9636.050.5707.253.5006.250.8607.453.9506.451.2607.654.2806.651.7807.854.5206.852.3658.054.6809.巴比妥钠-盐酸缓冲液（18℃）pH0.04M巴比妥钠/ml0.2M盐酸/mlpH0.04M巴比妥钠/ml0.2M盐酸/ml6.810018.48.41005.217.010017.88.61003.827.210016.78.81002.527.410015.39.01001.657.610013.49.21001.137.810011.479.41000.708.01009.399.61000.358.21007.21巴比妥钠分子量＝206.180.04M溶液为8.25g/L10.Tris-盐酸缓冲液50ml0.1M三羟甲基氨基甲烷(Tris)溶液于Xml0.1M盐酸混匀后，加水稀释至100mlpHX/mlpHX/ml7.145.78.126.27.244.78.222.97.343.48.319.97.442.08.417.27.540.38.514.77.638.58.612.47.736.68.710.37.834.58.88.57.932.08.97.08.029.2三羟甲基氨基甲烷(Tris)分子量＝121.140.1M溶液为12.114g/LTris溶液可以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/ml0.2M硼酸/mlpH0.05M硼砂/ml0.2M硼酸/ml7.41.09.08.23.56.57.61.58.58.44.55.57.82.08.08.76.04.08.03.07.09.08.02.0硼砂Na2B4O7·10H2O分子量＝381.430.05M(=0.2M硼酸根)溶液为19.07g/L硼酸H3BO3分子量＝61.840.2M溶液为12.37g/L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）Xml0.2M甘氨酸+Yml0.2M氢氧化钠再加水稀释至200mlpHX/mlY/mlpHX/mlY/ml8.6504.09.65022.48.8506.09.85027.29.0508.810.05032.09.25012.010.45038.69.45016.810.65045.5甘氨酸分子量＝75.070.2M甘氨酸溶液含15.01g/L13.硼砂-氢氧化钠缓冲液（0.05M硼酸根）Xml0.05M硼砂+Yml0.2M氢氧化钠再加水稀释至200mlpHX/mlY/mlpHX/mlY/ml9.3506.09.85034.09.45011.010.05043.09.65023.010.15046.0硼砂Na2B4O7·10H2O分子量＝381.430.05M(=0.2M硼酸根)溶液为19.07g/L14.碳酸钠-碳酸氢钠缓冲液（0.1M）Ca2+，Mg2+存在时不得使用pH0.1M碳酸钠/ml0.1M碳酸氢钠/ml20℃37℃9.168.77199.409.12289.519.40379.789.50469.909.725510.149.906410.2810.087310.5310288210.83105791Na2CO3·10H2O分子量＝286.20.1M溶液为28.62g/LNaHCO3分子量＝84.00.1M溶液为8.40g/L