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Non-volatile and Volatile Buffer Systems

Non-volatile buffers for anion exchange chromatography

Non-volatile buffers for anion exchange chromatography
pH intervalSubstanceConc. (mM)Counter-ionpKa
(25 °C)1
d(pKa)/
dT (°C)
4.3–5.3N-Methylpiperazine         20Cl-4.75-0.015
4.8–5.8Piperazine 20Cl- or HCOO-5.33-0.015
5.5–6.5L-Histidine20Cl-6.04 
6.0–7.0bis-Tris 20Cl-6.48-0.017
6.2–7.2; 8.6–9.6bis-Tris propane 20Cl-6.65; 9.10 
7.3–8.3Triethanolamine 20Cl- or CH3COO-7.76-0.020
7.6–8.6Tris 20Cl-8.07-0.028
8.0–9.0N-Methyldiethanolamine 20SO42-8.52-0.028
8.0–9.0N-Methyldiethanolamine 50Cl- or CH3COO-8.52-0.028
8.4–9.4Diethanolamine20 at pH 8.4
50 at pH 8.8
Cl-8.88-0.025
8.4–9.4Propane 1,3-Diamino     20Cl-8.88 -0.031
9.0–10.0Ethanolamine 20Cl-9.50-0.029
9.2–10.2Piperazine 20Cl-9.73-0.026
10.0–11.0 Propane 1,3-Diamino       20Cl-10.55-0.026
10.6–11.6Piperidine 20Cl-11.12-0.031
1Ref: Handbook of chemistry and physics, 83rd edition, CRC, 2002–2003.

Non-volatile buffers for cation exchange chromatography

Non-volatile buffers for cation exchange chromatography
pH intervalSubstanceConc. (mM)Counter-ionpKa
(25 °C)1
d(pKa)/
dT (°C)
1.4–2.4Maleic acid20Na+1.92 
2.6–3.6Methyl malonic acid20Na+ or Li+3.07 
2.6–3.6Citric acid20Na+3.13-0.0024
3.3–4.3 Lactic acid50Na+3.86 
3.3–4.3Formic acid50Na+ or Li+3.75+0.0002
3.7–4.7; 5.1–6.1 Succinic acid50Na+4.21; 5.64-0.0018
4.3–5.3Acetic acid50Na+ or Li+4.75+0.0002
5.2–6.2 Methyl malonic acid50Na+ or Li+5.76 
5.6–6.6MES50Na+ or Li+6.27-0.0110
6.7–7.7 Phosphate50Na+7.20-0.0028
7.0–8.0HEPES50Na+ or Li+7.56-0.0140
7.8–8.8BICINE50Na+8.33-0.0180
1Ref: Handbook of chemistry and physics, 83rd edition, CRC, 2002–2003.

Volatile buffer systems

pH rangBuffer systemCounter-ionpKa-values for buffering ions1
3.3–4.3Formic acid H+3.75
3.3–4.3; 4.8–5.8 Pyridine/formic acidHCOO-3.75; 5.25
3.3–4.3; 9.3–10.3Trimethylamine/formic acid HCOO-4.75; 9.81
4.3–5.8 Pyridine/acetic acidCH3COO-4.75; 5.25
4.3–5.3; 9.3–10.3Trimethylamine/acetic acid CH3COO-4.75; 9.81
3.3–4.3; 8.8–9.8 Ammonia/formic acidHCOO-3.75; 9.25
4.3–5.3; 8.8–9.8Ammonia/acetic acid CH3COO-4.75; 9.25
5.9–6.9; 9.3–10.3 Trimethylamine/carbonateCO32-6.35; 9.81
5.9–6.9; 8.8–9.8Ammonium bicarbonateHCO3-6.35; 9.25
5.9–6.9; 8.8–9.8 Ammonium carbonate/ammoniaCO32-6.35; 9.25
5.9–6.9; 8.8–9.8Ammonium carbonateCO32-6.35; 9.25
4.3–5.3: 7.2–8.2 N-ethylmorpholine/acetateHCOO-4.75; 7.72
1Ref: Handbook of chemistry and physics, 83rd edition, CRC, 2002–2003.
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