标准缓冲液的配制方法.docx

1、标准缓冲液的配制方法标准缓冲液的配制及常用数据一、标准缓冲液pH值与温度对照表温度四草酸氢钾0.05M邻苯二甲酸氢钾0.05M混合磷酸盐0.025M硼砂0.01M51.674.006.959.39101.674.006.029.33151.674.006.909.28201.684.006.889.23251.684.006.869.18301.684.016.859.14351.694.026.849.11二、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0.2 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.

2、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/L）X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)XY2.22.42.62.83.0555554.0703.9603.2952.6422.0223.23.43.63.855551.4700.9900.5970.263邻苯二甲酸氢钾分子量 = 204.23，0.2 mol/L邻苯二甲酸

3、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）2.22.42.62.83.03.23.43.63.84.04.24.44.64.85.00.401.242.183.174.114.945.706.447.107.718.288.829.359.8610.3010.6018.7617.8216.8315.8915.0614.3013.5612.9012.2911.7211.1810.6510.149.705.25.45.65.86.06.26.46

4、.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。Na2HPO4-2H2O分子量 = 178.05，0.2 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

5、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（0.1 mol/L）pH0.1 mol/L柠檬酸（毫升）0.1 mol/L柠檬酸钠（毫升）pH0.1 mo

6、l/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210.14，0.1 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

7、液为29.41克/毫升。6乙酸乙酸钠缓冲液（0.2 mol/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)2.63.84.04.24.44.60.751.201.802.653.704.909.258.808.207.356.305.104.85.05.25.45.65.85.907.007.908.609.109.404.103.002.101.400.900.60Na2Ac3H2O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

8、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/L Na2HPO4(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

9、.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35.61克/升。Na2HPO412H2O分子量 = 358.05，0.2 mol/L溶液为71.61克/升。NaH2PO42H2O分子量 = 156.01，0.2 mol/L溶液为31.202克/升。（2）磷酸氢二钠磷酸二氢钾缓冲液（1/15 mol/L）pHmol/L/15Na2HPO4(毫升)mol/L/15KH2PO4(毫升)pHmol/L/15Na2HPO4(毫升)mol/L/15KH2PO4(毫升)4.925.295.

10、916.246.476.646.816.980.100.501.002.003.004.005.006.009.909.509.008.007.006.005.004.007.177.387.738.048.348.678.187.008.009.009.509.759.9010.003.002.001.000.500.250.100Na2HPO42H2O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（0.05M）X毫升0.2M K2PO4 + Y毫升0.2N NaOH加水稀释至2

11、9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

12、.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，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)7.107.207.307.407.507.607.707.807.908.0045.744.743.442.040.338.536.634.532.029.28.108.208.308.408.508.608.708.808.9026.222.91

13、9.917.214.712.410.38.57.0三羟甲基氨基甲烷（Tris）HOCH2 CH2OHCHOCH2 NH2分子量=121.14;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硼砂（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Na2B4O7H2O,分子量=381.43; 0.05M溶液含19.07克/升。硼酸H2BO3,

14、分子量=61.84, 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。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pH0.05M硼砂（毫升）0.2NNaOH（毫升）pH0.05M硼砂（毫升）0.2NNaOH（毫升）9.39.49.69.850.050.050.050.06.011.023.034.010.010.150.050.043.046.012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pHXYpHXY8.68.89.09.29.45050505050

15、4.06.08.812.016.89.69.810.010.410.6505050505022.427.232.038.645.5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Na2B4O710H2O,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pH0.1MNa2CO

16、3(毫升)0.1MN2HCO3(毫升)20379.169.409.519.789.9010.1410.2810.5310.838.779.129.409.509.729.9010.0810.2810.57123456789987654321Na2CO210H2O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PH7.67.47.27.0H2ONaClNa2HPO4NaH2PO410008.52.20.110008.52.20.210008.52.20.3100ml8.5g2.2g0.4g三、实验室中常用酸碱的比

17、重和浓度名称分子式分子量比重面分浓度%（W/W）当量浓度（粗略）N配1升1N溶液所需毫升数盐酸HCl36.471.1937.212.0841.1835.411.81.1020.06.0硫酸H2SO498.091.8495.636.0281.1824.86.0硝酸HNO263.021.4270.9816.0631.4065.314.51.2032.366.1冰乙酸CH3COOH60.051.0599.517.459乙酸CH3COOH98.06366.0磷酸H2PO435.051.7185.015,30,45(依反应而定)67(以15N计)氨水NH4OH0.9015670.90427.014.37

18、00.9125.013.40.9610.05.6氢氧化钠溶液NaOH40.01.550.01953四、硫酸铵饱和度的常用表1调整硫酸铵溶液饱和度计算表（25）硫整硫酸铵终浓度,%饱和度10202530333540455055606570758090100每1升溶液加固体硫酸铵的克数硫酸铵初浓度，%饱和度0561141441761962092432773133513904304725165616627671057861181371501832162512883263654064494945926942029597891123155190225262300340382424520619253049

19、619312515819323026730734839048558330193062941271621982352733143564495463312437410714217721425229233342652235316394129164200238178319411506403163971321682052452853754694532659913417121025033943150336610113717621430239255336710314117926435360346910514322731465347010719027570357215323775361151988077157

20、9079在25下，硫酸铵溶液由初浓度调到终浓度时，每升溶液所加固体硫酸铵的克数。2 调整硫酸铵溶液饱和度计算表（0）在0下，硫酸铵溶液由初浓度调到终浓度时，每100毫升溶液所加固体硫酸铵的克数。常见蛋白质分子量参考值 （单位：dalton）蛋白质分子量肌球蛋白myosin甲状腺球蛋白thyroglobulin-半乳糖苷酶-galactosidase副肌球蛋白paramyosin磷酸化酶aphosphorylase a血清白蛋白serum albuminL-氨基酸氧化酶L-amino acid oxidase地氧化氢酶catalase丙酮酸激活酶pyruvate kinase谷氨酸脱氢酶glut

21、amate dehydrogenase亮氨酸氨肽酶glutamae dehydrogenase-球蛋白，H链-globulin, H chain延胡索酸酶（反丁烯二酸酶）fumarase卵白蛋白ovalbumin醇脱氢酶（肝）alcohol dehydrogenase (liver)烯醇酶enolase醛缩酶aldolase肌酸激酶creatine kinase胃蛋白酶原pepsinogenD-氨基酸氧化酶D-amino acid oxidase醇脱氢酶（酵母）alcohol dehydrogenase (yeast)甘油醛磷酸脱氢酶dlyceraldehyde phosphate dehydrogenase原肌球蛋白tropomyosin乳酸脱氢酶lactate dehydrgenase胃蛋白酶pepsin转磷酸核糖基酶phosphoribosyl transferase天冬氨酸氨甲酰转移酶，C链aspe