Caffeine
Caffeine, potassium sorbate, and sodium benzoate are common additives used in the production of soft drinks, energy drinks, and various food products. Accurate quantification of these compounds is crucial for quality control and regulatory compliance in the beverage industry. High-Performance Liquid Chromatography (HPLC) provides a robust method for the analysis of caffeine, potassium sorbate, and sodium benzoate mixtures. In this study, we describe the HPLC analysis of this mixture utilizing an Amaze HA mixed-mode column. Caffeine is a naturally occurring alkaloid found in coffee beans, tea leaves, and cacao pods. It is a white, crystalline powder with a bitter taste and acts as a stimulant, enhancing alertness and reducing fatigue. Potassium sorbate is a potassium salt of sorbic acid, a compound naturally found in some berries. It is widely used as a preservative in food and beverage products due to its ability to inhibit the growth of molds, yeasts, and bacteria. Sodium benzoate is the sodium salt of benzoic acid, a common food preservative found naturally in fruits. It is utilized in the food and beverage industry to prevent microbial growth and extend shelf life. The Amaze HA mixed-mode column features a unique stationary phase that combines hydrophobic, anion-exchange and cation-exclusion properties. The stationary phase consists of hydrophobic alkyl chains with embedded hydroxyl and amine groups, providing both reverse-phase and ion interactions. This mixed-mode chromatography enables enhanced selectivity and resolution of analytes, making it suitable for the analysis of polar and non-polar compounds, such as caffeine, potassium sorbate, and sodium benzoate. The Amaze HA column offers exceptional versatility and robustness, making it well-suited for various applications in the food and beverage industry. The HPLC analysis of a mixture of caffeine, potassium sorbate, and sodium benzoate on the Amaze HA mixed-mode column provides an effective method for the quantitative determination of these compounds commonly used in soft drinks, energy drinks, and other food products. This approach offers high sensitivity, selectivity, and reproducibility, making it suitable for quality control and regulatory compliance. Moreover, the unique design and superior performance of the Amaze HA mixed-mode column ensure efficient separation and analysis of complex mixtures, further enhancing its utility in beverage analysis and research endeavors.
Column: | Amaze HA |
Separation Modes: | reversed-phase, Anion-exchange |
Column Dimenstions: | 3x100 mm, 3 um, 100 A |
Mobile Phase: | ACN/water/ammonium formate |
Detection: | UV 255 nm, ELSD |
Sample: | various-various mg/ml |
Injection: | 3 uL |
Flow rate: | 0.6 ml/min |
Class of compounds: | Aromatic acid, Inorganic cation, Organic acid, Preservative, Stimulant, Xanthine |
Nature of compounds: | Acidic, Basic, Hydrophilic, Neutral, Polar |
Compounds: | Sodium ion, Potassium ion, Caffeine, Sorbic acid, Benzoic acid |
Column: | Amaze C18 |
Separation Modes: | reversed-phase |
Column Dimenstions: | 4.6x150 mm, 3 um, 100A |
Mobile Phase: | 25% ACN with 50 mM ammonium phosphate and 0.1% phosphoric acid |
Detection: | 220 nm |
Sample: | 0.3-0.5 mg/ml |
Injection: | 1 uL |
Flow rate: | 1 ml/min |
Class of compounds: | Aromatic acid, Aromatic compound, Drug, Stimulant, Xanthine |
Nature of compounds: | Hydrophilic, Hydrophobic, Neutral, Polar |
Compounds: | Acetaminophen, Caffeine, Aspirin |
Comparison run on Amaze PFP, Amaze AR, Amaze SPF, and Heritage C20 columns.
Column: | Amaze AR |
Separation Modes: | reversed-phase, pi-pi |
Column Dimenstions: | 4.6x150 mm, 5 μm, 100A |
Mobile Phase: | 28% ACN with 20 mM AmAc pH 4.5 |
Detection: | UV 255 nm |
Sample: | 1 mg/ml |
Injection: | 3 μL |
Flow rate: | 1 ml/min |
Class of compounds: | Amino acid, Aromatic acid, Aromatic compound, Drug, Isomer, Organic acid, Preservative, Stimulant, Xanthine |
Nature of compounds: | Acidic, Hydrophilic, Hydrophobic, Neutral, Polar, Zwitterionic |
Compounds: | Caffeine, 4-Aminobenzoic acid, Aspirin, 4-Hydroxybenzoic acid, Sorbic acid, Benzoic acid |
Column: | Heritage С18 |
Separation Modes: | reversed-phase |
Column Dimenstions: | 4.6 x 150 mm, 5 um, 100A |
Mobile Phase: | NaFm/H2O/MeCN/THF 1.64 g /2L/50 ml/40 ml/ pH 4.5 with acetic acid |
Detection: | UV 270 nm |
Sample: | 0.2 mg/ml |
Injection: | 5 uL |
Flow rate: | 1 mL/min |
Class of compounds: | Stimulant, Xanthine |
Nature of compounds: | Hydrophilic, Neutral |
Compounds: | Caffeine |
Column: | Heritage С18 |
Separation Modes: | reversed-phase |
Column Dimenstions: | 4.6 x 100 mm, 5 um, 100A |
Mobile Phase: | MeOH/H2O/AcOH 30/67/3 |
Detection: | UV 275 nm |
Sample: | 0.2 mg/ml |
Injection: | 5 uL |
Flow rate: | 1 mL/min |
Class of compounds: | Aromatic acid, Aromatic compound, Drug, Organic acid, Stimulant, Xanthine |
Nature of compounds: | Acidic, Hydrophilic, Hydrophobic, Neutral, Polar |
Compounds: | Acetaminophen, Caffeine, Aspirin, Benzoic acid, Salicylic acid |
Cytosine is one of four basic compounds that play an important role in building DNA and RNA molecules. Cytidine consist of cytosine and ribose molecules. Cytosine and cytidine molecules are polar weak basis. They have an ability to form hydrogen bonds. Order of elution and selectivity of separation can be changed by changing mobile phase composition. Method is compatible with most common HPLC detection techniques: MS, RI, UV, ELSD, CAD, etc.
Column: | Amaze HD |
Separation Modes: | HILIC |
Column Dimenstions: | 3.2 x 50 mm |
Mobile Phase: | MeCN/MeOH (95/5) with 0.1% HCOOH with 0.01%AmFm |
Detection: | UV 275 nm |
Sample: | 0.3 mg/ml |
Injection: | 1 uL |
Flow rate: | 1.0 mL/min |
Class of compounds: | Stimulant, Xanthine |
Nature of compounds: | Hydrophilic, Neutral, Polar |
Compounds: | Caffeine, 3-Methylxanthine, 1-Methylxanthine, Xanthine |
Core-shell mixed-mode columns offer unique selectivity which comes from the presence of two mechanisms - reversed-phase and ion-exchange. The alternative selectivity of mixed-mode columns can be used when order of elution or resolution between compounds needs improvement. Even for neutral compounds a different selectivity can be obtained. A method for common components of headache medications (acetaminophen, caffeine, and benzoate) was developed on our Coresep 100 column. Efficiency and peak shape are comparable to leading brand core-shell reversed-phase columns.
Column: | Coresep 100 |
Separation Modes: | reversed-phase |
Column Dimenstions: | 3.2 x 50 mm, 2.7 um, 90A |
Mobile Phase: | 20% ACN with 0.1% TFA |
Detection: | UV 250 nm, ELSD |
Sample: | 0.3 mg/ml |
Injection: | 1 uL |
Flow rate: | 1 ml/min |
Class of compounds: | Aromatic acid, Aromatic compound, Drug, Organic acid, Stimulant, Xanthine |
Nature of compounds: | Acidic, Hydrophilic, Neutral |
Compounds: | Acetaminophen, Caffeine, Benzoic acid |