Bio-rad UNO® Monolith Cation Exchange Columns Uživatelský manuál stáhnout pdf (Strana 4)

while maintaining maximum buffering capacity. In any case, a buffer concentration

of 20 mM is recommended.

As can be seen in Table 2 and 3, the pK

and hence the pH of the buffer, changes

with temperature. Therefore the pH of the buffer must be adjusted at the working

temperature.

Table 2. Buffers for Anion-Exchange Chromatography

pH range Buffer Mwt pK

@25 °C Counter-ion ∆ pK

/°C

5.0 - 6.0 Piperazine 86.1 5.7 Cl

/HCOO- -0.015

5.5 - 6.0 L-Histidine 155.2 6.15 Cl

5.8 - 7.2 Bis-Tris 209.2 6.5 Cl

-0.017

6.4 - 7.3 Bis-Tris Propane 282.3 6.8, 9.0 Cl

7.3 - 8.3 Triethanolamine 149.2 7.8 Cl

/CH

COO

-0.020

7.6 - 8.6 Tris 121.1 8.1 Cl

-0.031

8.4 - 8.8 Diethanolamine 105.1 8.9 Cl

-0.025

9.0 - 9.9 Ethanolamine 61.1 9.5 Cl

-0.029

9.8 - 10.3 1,3-diamino-propane 74.1 10.5 Cl

-0.026

Table 3. Buffers for Cation-Exchange Chromatography

pH range Buffer Mwt pK

@25 °C Counter-ion ∆ pK

/°C

3.6 - 4.3 Lactic acid 90.1 3.8 Na

4.2 - 5.2 Citric acid 192.1 3.1 Na

5.5 - 6.7 MES 195.2 6.1 Na

-0.011

6.1 - 7.5 PIPES 302.4 6.8 Na

-0.009

6.5 - 7.9 MOPS 209.3 7.2 Na

-0.006

6.7 - 7.6 Phosphate 120 (Monobasic) 7.2 Na

-0.003

142 (Dibasic)

6.8 - 8.2 TES 229.2 7.4 Na

-0.020

6.8 - 8.2 HEPES 238.3 7.5 Na

-0.014

7.4 - 8.8 Tricine 179.2 8.1 Na

-0.021

Always use buffer components of the highest purity available as UV-absorbing

impurities may cause baseline disturbances and interfere with the detection of pro-

tein peaks.

4006078A 10/1/98 10:02 AM Page 3

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