Caffeine

HPLC Analysis of Mixture of Caffeine, Potassium Sorbate and Sodium Benzoate on Amaze HA Mixed-Mode Column
Application description

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.

Conditions of Experiment
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
Analytes
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
HPLC Analysis of Excedrin Pain Medication on Reversed-Phase Aromatic Phases
Conditions of Experiment
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
Analytes
Class of compounds: Aromatic acid, Aromatic compound, Drug, Stimulant, Xanthine
Nature of compounds: Hydrophilic, Hydrophobic, Neutral, Polar
Compounds: Acetaminophen, Caffeine, Aspirin
HPLC Analysis of 6 Aromatic Acidic, Zwitterionic and Neutral Compounds on Amaze Aromatic Stationary Phases
Application description

Comparison run on Amaze PFP, Amaze AR, Amaze SPF, and Heritage C20 columns.

Conditions of Experiment
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
Analytes
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
HPLC Analysis of Caffeine on Heritage C18 Column According to US Pharmacopeia
Conditions of Experiment
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
Analytes
Class of compounds: Stimulant, Xanthine
Nature of compounds: Hydrophilic, Neutral
Compounds: Caffeine
HPLC Analysis of Drug Composition on Heritage C18 Column According to US Pharmacopeia
Conditions of Experiment
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
Analytes
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
HPLC Method for Analysis of Caffeine and Other Xanthines on Amaze HD Column
Application description

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.

Conditions of Experiment
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
Analytes
Class of compounds: Stimulant, Xanthine
Nature of compounds: Hydrophilic, Neutral, Polar
Compounds: Caffeine, 3-Methylxanthine, 1-Methylxanthine, Xanthine
Selectivity Difference for Polar Compounds in Reversed-Phase and Mixed-Mode Separation on Coresep 100 HPLC Column
Application description

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.

Conditions of Experiment
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
Analytes
Class of compounds: Aromatic acid, Aromatic compound, Drug, Organic acid, Stimulant, Xanthine
Nature of compounds: Acidic, Hydrophilic, Neutral
Compounds: Acetaminophen, Caffeine, Benzoic acid