常用緩沖液配方

1、常見緩沖液的緩沖范圍Tris-鹽酸緩沖液(0.05mol/L25 C )甘氨酸-氫氧化鈉緩沖液(0.05mol/L) 硼砂-氫氧化鈉緩沖液(0.05mol/L) 碳酸鈉-碳酸氫鈉緩沖液(0.1mol/L) 碳酸鈉-氫氧化鈉緩沖液(0.025mol/L) 磷酸氫二鈉-氫氧化鈉緩沖液 氯化鉀-鹽酸緩沖液(0.2mol/L ) 氯化鉀-氫氧化鈉緩沖液(0.2mol/L ) 參考下圖：常見緩沖液的緩沖范圍7.10-9.008.6-10.69.3-10.19.16-10.839.6-11.010.9-12.01.0-2.212.0-13.0三羥甲基氨基甲烷 (Tris) Mr=121.14 甘氨酸Mr=

2、75.07 硼砂 Na2B4O7?10H2O Mr=381.43 碳酸鈉Mr=286.2碳酸氫鈉 Mr=84.0氯化鉀Mr=74.55氯化鉀Mr=74.55緩沖液名稱及常用濃度MES (2嗎咻代乙磺酸)緩沖pH范圍5.5-6.7主要物質(zhì)分子量195.2Bis-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-鹽酸緩沖液(0.05mol/L )2.2-5.0甘氨酸Mr=75.07鄰苯二甲酸-鹽酸緩沖液(0.05mol/L )

3、2.2-3.8鄰苯二甲酸氫鉀 Mr=204.23磷酸氫二鈉-檸檬酸緩沖液2.2-8.0磷酸氫二鈉 Mr=141.98檸檬酸-氫氧化鈉-鹽酸緩沖液2.2-6.5檸檬酸 Mr=192.06檸檬酸-檸檬酸鈉緩沖液(0.1mol/L ) 3.0-6.6檸檬酸Mr=192.06檸檬酸鈉Mr=257.96乙酸-乙酸鈉緩沖液(0.2mol/L )3.6-5.8乙酸鈉 Mr=81.76乙酸 Mr=60.05鄰苯二甲酸氫鉀-氫氧化鈉緩沖液4.1-5.9氫鉀 Mr=204.23磷酸氫二鈉-磷酸二氫鈉緩沖液(0.2mol/L )5.8-8.0磷酸氫二鈉-磷酸二氫鉀緩沖液(1/15 molL ) 4.92-8.18磷

4、酸二氫鉀-氫氧化鈉緩沖液(0.05mol/L )5.8-8.0巴比妥鈉-鹽酸緩沖液(18 C)6.8-9.6巴比妥鈉Mr=206.18參考下圖：常見緩沖液的緩沖范圍Buffering 沖卿 of comnwttty iwetf buffer (»t 30*C)USflLS7UBflID榆患1111J1四fl.lADA輸PIPES63MOPSO«.971MOPSTES74rtPfS7SL_HUSS.liEPP3L8JDnTfilCINEa.F|W："TAPS9.4ROW S.4CM牯 9.3 3S m常用緩沖液的配制方法1. 甘氨酸-鹽酸緩沖液(0.05M)X ml

5、 0.2M 甘氨酸 +Y ml 0.2M 鹽酸 再加水稀釋至 200mlpHX/mlY/mlpHX/ml Y/ml2.25044.0 3.05011.42.45032.4 3.2508.22.65024.2 3.4506.42.85016.8 3.6505.0甘氨，酸分子量=75.07 0.2M甘氨酸溶液含15.01g/L2. 鄰苯二甲酸-鹽酸緩沖液(0.05M )X ml 0.2M 鄰苯二甲酸氫鉀 +Y ml 0.2M 鹽酸 再加水稀釋至 200mlpHXY pH XY2.254.670 3.25 1.4702.453.960 3.45 0.9902.653.295 3.65 0.5972.

6、852.642 3.85 0.2633.052.032鄰苯二甲酸氫鉀分子量=2.4.23 0.2M鄰苯二甲酸氫鉀溶液含40.85g/L3. 磷酸氫二鈉-檸檬酸緩沖液pH 0.2M Na2HPO4/ml 0.1M 檸檬酸 /ml pH 0.2M Na2HPO4/ml 0.1M 檸檬酸 /ml2.2 0.40 19.6 5.2 10.72 9.282.4 1.24 18.76 5.4 11.15 8.852.62.1817.825.611.608.402.83.1716.835.812.097.913.04.1115.896.012.637.373.24.9415.066.213.226.783.

7、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=141.98 0.2M 溶液含 28.40g/L Na2HPO4 - 2H2O 分子量=178.05 0.2M 溶液含 35.61g/L C6H8O

8、7 - H2O 分子量=210.14 0.1M 溶液含 21.01g/L4.檸檬酸-氫氧化鈉-鹽酸緩沖液pH鈉離子濃度/M檸檬酸 C6H8O7 - H2O/g氫氧化鈉NaOH/g濃鹽酸HCl/ml終體積/L2.20.20 210 84160 103.10.20 210 83116 103.30.20 210 83106 104.30.20 210 8345 105.30.35 245 14468 105.80.45 285 186105 106.50.38 266 156126 10使用時可以每升中加入 1g酚，若最后pH有變化，再用少量 50%氫氧化鈉溶液或濃鹽酸調(diào) 節(jié)，冰箱保存。5. 檸檬

9、酸-檸檬酸鈉緩沖液(0.1M ) pH 0.1M檸檬酸/ml 0.1M檸檬酸鈉/ml pH 0.1M檸檬酸/ml 0.1M檸檬酸鈉/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=210.14 0.1M 溶液含 21.01g/L

10、Na3C6H5O7 - 2H2O 分子量=294.12 0.1M 溶液含 29.41g/L6. 乙酸-乙酸鈉緩沖液(0.2M)pH 0.2M NaAc/ml 0.2M HAC/ml pH 0.2M NaAc/ml 0.2M HAC/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=136.09 0.2M 溶液為 27.22g/L7. 磷酸鹽緩沖液(1)磷酸氫二鈉

11、-磷酸二氫鈉緩沖液(0.2M)pH 0.2M Na2HPO4/ml 0.2M NaH2PO4/ml pH 0.2MNa2HPO4/ml 0.2M NaH2PO4/ml5.88.092.07.061.039.05.910.090.07.167.033.06.012.387.77.272.028.06.115.085.07.377.023.06.218.581.57.481.019.06.322.577.57.584.016.06.426.573.57.687.013.06.531.568.57.789.510.56.637.562.57.891.58.56.743.556.57.993.07.0

12、6.849.551.08.094.75.36.955.045.0Na2HPO4 - 2H2O 分子量=178.05 0.2M 溶液含 35.61g/LNa2HPO4 - 12H2O 分子量=358.22 0.2M 溶液含 71.64g/LNaH2PO4 - H2O 分子量=138.01 0.2M 溶液含 27.6g/LNaH2PO4 - 2H2O 分子量=156.03 0.2M 溶液含 31.21g/L磷酸氫二鈉-磷酸二氫鉀緩沖液(1/15M )pH 1/15M Na2HPO4/ml 1/15M KH2PO4/ml pH 1/15MNa2HPO4/ml 1/15M KH2PO4/ml4.920

13、.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=178.05 1/15M 溶液含 35.61g/LKH2PO4 分子量=136.09 1/15M 溶液含 9.078g/L8. 磷酸二氫鉀-氫氧化鈉緩沖液(0.05M)X ml 0.2M KH2PO4 +Y ml 0

14、.2M NaOH再加水稀釋至 20mlpH(20 C) X/ml Y/ml pH(20 C) X/ml Y/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.巴比妥鈉-鹽酸緩沖液(18C)pH0.04M巴比妥鈉/ml0.2M 鹽酸 /mlpH0.04M巴比妥鈉/ml0.2M 鹽酸 /ml6.810018.48.41005.217.010017.88.61003.827.210016.78.81002.527.410015.

15、39.01001.657.610013.49.21001.137.810011.479.4100 0.708.01009.399.61000.358.21007.21100巴比妥鈉分子量=206.18 0.04M溶液為8.25g/L10.Tris-鹽酸緩沖液50ml 0.1M三羥甲基氨基甲烷(Tris)溶液于X ml 0.1M鹽酸混勻后，加水稀釋至100mlpHX/mlpHX/ml7.145.78.126.27.244.78.222.97.343.48.319.97.442.08.417.27.540.38.514.77.638.58.612.47.736.68.710.37.834.58.8

16、8.57.932.08.97.08.029.2三羥甲基氨基甲烷(Tris)分子量=121.14 0.1M溶液為12.114g/LTris溶液可以從空氣中吸收二氧化碳，使用時注意將瓶蓋嚴11.硼酸-硼砂緩沖液(0.2M硼酸根)pH0.05M 硼砂 /ml0.2M 硼酸 /ml pH0.05M 硼砂 /ml0.2M 硼酸 /ml7.41.0 9.0 8.23.5 6.57.61.5 8.5 8.44.5 5.57.82.0 8.0 8.76.0 4.08.03.0 7.0 9.08.0 2.0硼砂Na2B4O7 - 10H2O 分子量=381.43 0.05M（=0.2M 硼酸根）溶液為 19.0

17、7g/L硼酸H3BO3分子量=61.84 0.2M 溶液為12.37g/L硼砂易失去結晶水，必須在帶塞的瓶中保存12. 甘氨酸-氫氧化鈉緩沖液(0.05M )X ml 0.2M 甘氨酸+Y ml 0.2M 氫氧化鈉 再加水稀釋至 200mlpH8.6X/ml Y/ml pH X/ml Y/ml50 4.0 9.6 50 22.48.850 6.0 9.8 50 27.29.050 8.8 10.0 50 32.09.250 12.0 10.4 50 38.69.450 16.8 10.6 50 45.5甘氨酸分子量=75.07 0.2M甘氨酸溶液含15.01g/L13. 硼砂-氫氧化鈉緩沖液(0.05M硼酸根)X ml 0.05M 硼砂+Y ml 0.2M 氫氧化鈉 再加水稀釋至 200mlpHX/mlY/ml pH X/ml Y/ml9.3506.0 9.8 50 34.09.45011.0 10.0 50 43.09.65023.0 10.1 50 46.0硼砂Na2B4O7 - 10H2O 分子量=381.43 0.05M（=0.2