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1、.常用缓冲溶液的配制方法1甘氨酸盐酸缓冲液0.05mol/LX毫升0.2 mol/L甘氨酸+Y毫升0.2 mol/L HCI，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 = 75.07，0.2 mol/L甘氨酸溶液含15.01克/升。2邻苯二甲酸盐酸缓冲液0.05 mol/LX毫升0.2 mol/L邻苯二甲酸氢钾 + 0.2 mol/L HCl，再加水稀释到20毫升pH(20)XYpH(20)XY2.22.42.62.83.0555554.0703.
2、9603.2952.6422.0223.23.43.63.855551.4700.9900.5970.263邻苯二甲酸氢钾分子量 = 204.23，0.2 mol/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/L Na2HPO4毫升0.1mol/L柠檬酸毫升pH0.2mol/L Na2HPO4毫升0.1mol/L柠檬酸毫升2.22.42.62.83.03.23.43.63.84.04.24.44.64.85.00.401.242.183.174.114.945.706.447.107.718.288.829.359.8610.3010.6018.7617.821
3、6.8315.8915.0614.3013.5612.9012.2911.7211.1810.6510.149.705.25.45.65.86.06.26.46.66.87.07.27.47.67.88.010.7211.1511.6012.0912.6313.2213.8514.5515.4516.4717.3918.1718.7319.1519.459.288.858.407.917.376.786.155.454.553.532.611.831.270.850.55Na2HPO4分子量 = 14.98，0.2 mol/L溶液为28.40克/升。Na2HPO42H2O分子量 = 178.0
4、5，0.2 mol/L溶液含35.01克/升。C4H2O7H2O分子量 = 210.14，0.1 mol/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C6H8O7H2O氢氧化钠(克)NaOH 97%盐酸毫升HCl浓最终体积升2.23.13.34.35.35.86.50.200.200.200.200.350.450.3821021021021024528526684838383144186156160116106456810512610101010101010使用时可以每升中参加1克克酚，假设最后pH值有变化，再用少量50%氢氧化钠溶液或浓盐酸调节，冰
5、箱保存。5柠檬酸柠檬酸钠缓冲液0.1 mol/LpH0.1 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C6H8O7H2O：分子量210.14，0
6、.1 mol/L溶液为21.01克/升。柠檬酸钠Na3 C6H5O72H2O：分子量294.12，0.1 mol/L溶液为29.41克/毫升。6乙酸乙酸钠缓冲液0.2 mol/LpH(18)0.2 mol/L NaAc(毫升)0.3 mol/L HAc(毫升)pH(18)0.2 mol/LNaAc(毫升)0.3 mol/LHAc(毫升)2.63.84.04.24.44.60.751.201.802.653.704.909.258.808.207.356.305.104.85.05.25.45.65.85.907.007.908.609.109.404.103.002.101.400.900.6
7、0Na2Ac3H2O分子量 = 136.09，0.2 mol/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0.2pH0.2 mol/L Na2HPO4(毫升)0.2 mol/L NaH2PO4(毫升)pH0.2 mol/LNa2HPO4(毫升)0.2 mol/L NaH2PO4(毫升)5.85.96.06.16.26.36.46.56.66.76.86.98.010.012.315.018.522.526.531.537.543.549.555.092.090.087.785.081.577.573.568.562.5 56.551.045.07.07.17.27.37
8、.47.57.67.77.87.98.061.067.072.077.081.084.087.089.591.593.094.739.033.028.023.019.016.013.010.58.57.05.3Na2HPO42H2O分子量 = 178.05，0.2 mol/L溶液为85.61克/升。Na2HPO412H2O分子量 = 358.14，0.2 mol/L溶液为71.628克/升。NaH2PO42H2O分子量 = 156.01，0.2 mol/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
9、X围最宽：NaH2PO4： pKa12.12，pKa27.21； Na2HPO4：pKa17.21，pKa212.32配酸性缓冲液：用NaH2PO4，pH14，配中性缓冲液：用混合的两种磷酸盐，pH68，配碱性缓冲液：用Na2HPO4，pH1012。用钾盐比钠盐好，因为低温时钠盐难溶，钾盐易溶，但假设配制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：容易配制成各种浓度的缓冲液；适用的pH X围宽；pH 受温度的影响小；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：
10、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；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1/15 mol/LpHmol/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
11、.009.009.509.759.9010.003.002.001.000.500.250.100Na2HPO42H2O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0.05MX毫升0.2M K2PO4 + Y毫升0.2N NaOH加水稀释至29毫升 pH(20)X (毫升)Y毫升pH(20)X (毫升)Y毫升5.86.06.26.46.66.85555550.3720.5700.8601.2601.7802.3657.07.27.47.67.88.05555552.9633.50
12、03.9504.2804.5204.6809巴比妥钠-盐酸缓冲液18pH0.04M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10Tris盐酸缓冲液0.05M，2550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。
