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Organic Acids
The analysis of organic acids is of great importance in many areas of research and industry. For example, in the analysis of wine, beer or fruit juices or in connection with the analysis of carbohydrates.
An important chromatographic separation method for analysing organic acids is ion exclusion chromatography (IEC). Polymer resins with sulphonic acid groups (-SO3H) on the surface are usually used as the stationary phase. Due to a weakly acidic mobile phase, the sulphonic acid groups are partially deprotonated, whereby the surface is negatively charged towards the outside. Depending on the mobile phase and its pKa value, the analytes are more or less in the following equilibrium: R-COO-+ H+ ↔ R-COOH.
Dissociated analytes can therefore not penetrate the pore system of the stationary phase and are excluded, whereas analytes in the undissociated state are retarded. Separation is therefore largely based on the pKa value of the analytes. Ion-exclusion chromatography can also be used for the simultaneous separation of sugars and organic acids.
Applications
Other common chromatographic separation methods for organic acids are reversed phase chromatography (RP-HPLC) and hydrophilic interaction chromatography (HILIC).
Ion Exclusion Separation of eleven Organic Acids with Concise Coregel 87H3
Peak identities
1. lactic acid 2. formic acid 3. acetic acid 4. propionic acid 5. isobutyric acid 6. butyric acid 7. isovaleric acid 8. valeric acid 9. isocaproic acid 10. caproic acid 11. heptanoic acid
Test conditions
Column: Coregel 87H3 300x7.8mm, 9µm
Mobile phase: 8 nM H2SO4
Flow rate: 0.6 mL/min
Temperature: 80 °C, 65 °C, 50 °C
Reversed Phase Analysis of Organic Acids with GL Sciences Inertsil ODS-4
Peak identities
1. malic acid 2. lactic acid 3. acetic acid 4. maleic acid 5. citric acid 6. succinic acid 7. fumaric acid
Test conditions
Column: Inertsil ODS-4 250x4.6mm, 5µm
Mobile phase: 20 mM NH4H2PO4, pH 2
Flow rate: 1 mL/min
Temperature: 40 °C
Detection: UV, 210 nm
Injection volume: 5 µL
HILIC Separation of Organic Acids with Imtakt Unison UK-Amino
Peak identities
1. succinic acid 2. lactic acid 3. sodium gluconate 4. malic acid 5. glyoxylic acid 6. fumaric acid 7. malonic acid 8. citric acid 9. tartaric acid 10. maleic acid
Test conditions
Column: Union UK-Amino 250x3mm, 3µm
Mobile phase A: 30/70/0.1 water/acetonitrile/formic acid
Mobile phase B: 100/2 water/formic acid
Gradient: 10-60%B in 25min, 100%B for 25 to 60min
Flow rate: 0.4 mL/min
Temperature: 37 °C
Detection: ELSD (spray chamber 190°C, drift tube 42 °C)
Injection volume: 10 µL
Downloads
You will find further information with some application examples in the following brochures:
Thermo-Dionex Acclaim Organic Acid Product Manual
Concise Transgenomic Coregel Organic Acid Analysis
Shodex Ion Exclusion Chromatography Columns
Sepax Carbomix Catalog
Sepax Retention-Time Table for Carbomix Phases
Manufacturer overview
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Avantor ACE
UHPLC - Reversed Phase:
- Avantor ACE Excel 1.7 C18 Amide, 1.7µm
HPLC - Reversed Phase:
- Avantor ACE Excel 3 C18 Amide, 3µm
- Avantor ACE 5 C18, 5µm
ChromaNik
HPLC - Reversed Phase:
- SunShell RP-AQUA, 2.6µm
Concise Separations
HPLC - Ion Exclusion:
- Coregel 87H3, 9µm
GL Sciences
HPLC - Reversed Phase:
- Inertsil ODS-4, 5µm
- InertSustain AQ-C18, 5µm
- InertSustain C18, 5µm
- Inertsil C8, 5µm
In tact
HPLC - HILIC:
- Union UK-Amino, 3µm
Macherey-Nagel
HPLC - Reversed Phase:
- Nucleodur C18 Gravity-SB, 5µm
- Nucleodur PolarTec, 5µm
Osaka-Soda
HPLC - Reversed Phase:
- Capcell PAK C18 MG S5, 5µm
Sepax
HPLC - Ion Exclusion:
- Carbomix H-NP10 8%, 10µm
Shodex
HPLC - Ion Exclusion:
- RSpak KC-811, 6µm
HPLC - Reversed Phase:
- RSpak DE-413, 4µm
Thermo Fisher
HPLC - Reversed Phase:
- Acclaim Organic Acid, 3µm
- Acclaim Organic Acid, 5µm
Tosoh
HPLC - Reversed Phase:
- TSKgel ODS-100V, 5µm
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