Analysis of Chlorogenic acid in Vitamin C medicine

Importance of the Topic

Chlorogenic acid is a key polyphenolic antioxidant found in a variety of plant‐derived products and dietary supplements. Monitoring its concentration in vitamin C tablets serves both to verify label claims and to ensure product purity and efficacy in quality control laboratories.

Objectives and Study Overview

The primary goal of this work was to develop and demonstrate a straightforward, reproducible reversed‐phase high‐performance liquid chromatography (HPLC) method for quantifying chlorogenic acid in commercial vitamin C medicine. The approach emphasizes minimal sample preparation and an isocratic mobile phase to facilitate routine implementation.

Methodology

The procedure consists of two main steps:

• Sample Preparation: Ten vitamin C tablets were de-coated, finely pulverized, and 1 g of the powder was transferred into a 100 mL volumetric flask. Methanol was added, followed by ultrasonic extraction (300 W, 40 kHz) for 45 minutes. After cooling, the solution was diluted to volume with methanol–water, mixed, and filtered before injection.
• Chromatographic Conditions: Isocratic elution was carried out using an aqueous solution of 1 % glacial acetic acid (mobile phase A) and acetonitrile (mobile phase B) in a 94:6 ratio. The flow rate was set to 1.0 mL/min, column temperature at 30 °C, and injection volume 10 µL. Detection was performed by UV absorbance at 327 nm.

Used Instrumentation

• Column: Shim-pack GIS C18 (250 mm × 4.6 mm I.D., 5 µm; P/N 227-30106-08)
• Detector: UV detector set at 327 nm
• Autosampler Vials: LabTotal™ Vials for LC/LCMS (P/N 227-34001-01)
• Ultrasonic Bath: 300 W, 40 kHz

Main Results and Discussion

The chromatogram of the chlorogenic acid standard (30 µg/mL) showed a single well‐resolved peak, confirming method specificity. Analysis of vitamin C tablet extracts revealed chlorogenic acid at expected retention times, with no significant interferences. Peak symmetry and baseline stability indicated robust performance under the chosen isocratic conditions.

Benefits and Practical Applications

This method offers:

• High reproducibility and straightforward execution for routine QC labs
• Minimal solvent consumption via isocratic elution
• Sufficient sensitivity for detecting chlorogenic acid at pharmaceutically relevant levels
• Applicability to various dietary supplement matrices

Future Trends and Opportunities

Advancements that could enhance this workflow include:

• Transition to ultrahigh‐performance liquid chromatography (UHPLC) for reduced analysis time and solvent use
• Integration of mass spectrometric detection for improved selectivity and lower detection limits
• Implementation of green solvents or solvent‐free sample preparation techniques
• Automation of sample handling to increase throughput

Conclusion

The presented HPLC method provides a reliable, cost‐effective tool for routine determination of chlorogenic acid in vitamin C tablets. Its straightforward sample preparation and isocratic separation make it well suited for quality assurance and regulatory compliance in pharmaceutical and nutraceutical industries.