Application description
This method outlines the High-Performance Liquid Chromatography (HPLC) separation of ascorbic acid, citric acid, etidronic acid, and ethylenediaminetetraacetic acid (EDTA) utilizing an Amaze HA Mixed-Mode Column. Each compound's unique properties are considered for effective separation, ensuring accurate quantification.
Ascorbic Acid: Also known as vitamin C, ascorbic acid is a water-soluble antioxidant involved in various physiological processes. It possesses acidic properties, making it amenable to interaction with the stationary phase of the HPLC column.
Citric Acid: Widely found in citrus fruits, citric acid is a tricarboxylic acid that contributes to the sour taste of fruits. It is water-soluble and contains multiple acidic functional groups, allowing for interaction with the column's stationary phase.
Etidronic Acid: A bisphosphonate used in the treatment of osteoporosis, etidronic acid is water-soluble and contains phosphonic acid functional groups. Its unique structure influences its behavior during HPLC separation.
Ethylenediaminetetraacetic Acid (EDTA): A chelating agent commonly used in analytical chemistry, EDTA is water-soluble and forms complexes with metal ions. Its chelating properties present specific challenges and opportunities in chromatographic separation.
Column Properties:
Amaze HA Mixed-Mode Column: This column features a mixed-mode stationary phase, offering both hydrophobic and ionic interactions. The hydrophobic component enables interaction with non-polar compounds, while the ionic component facilitates interactions with charged species. This versatility allows for effective separation of polar and ionizable compounds like ascorbic acid, citric acid, etidronic acid, and EDTA.
By employing this method, precise separation and quantification of ascorbic acid, citric acid, etidronic acid, and EDTA can be achieved, facilitating their analysis in pharmaceutical formulations, food products, and environmental samples.
Conditions of Experiment
| Column: |
Amaze HA |
| Separation Modes: |
Anion-exchange |
| Column Dimenstions: |
3x100 mm, 3 um, 100 A |
| Mobile Phase: |
50% ACN with 50 mM ammonium formate pH 3 |
| Detection: |
ELSD, 45*C |
| Sample: |
various-0.5 mg/ml |
| Injection: |
3 uL |
| Flow rate: |
0.6 ml/min |
Analytes
| Class of compounds: |
Aromatic acid, Drug, Organic acid, Preservative, Supplement, Vitamin |
| Nature of compounds: |
Acidic, Hydrophilic, Polar, Zwitterionic |
| Compounds: |
Etidronic acid, Citric acid, Ascorbic acid, Ethylenediaminetetraacetic acid |
