Analysis of Purine Bases

Význam tématu

In analytical chemistry, purine alkaloids such as caffeine, theophylline and theobromine play a critical role in food, pharmaceutical and biological studies. Accurate separation and quantification of these polar compounds demand robust methods that ensure high resolution and reproducibility.

Cíle a přehled studie

This study evaluates the performance of a Shim-pack GIST Amide column under hydrophilic interaction liquid chromatography (HILIC) conditions to separate eight purine bases and related compounds. Key objectives include optimizing mobile phase composition, assessing chromatographic resolution and demonstrating method suitability for routine analysis.

Použitá metodika

An isocratic HILIC method was employed using a ternary mobile phase of acetonitrile, water with 50 mmol/L ammonium formate and formic acid (90/10/0.1, v/v/v). The flow rate was set at 0.2 mL/min with the column maintained at 40 °C. Detection was performed at 254 nm, and sample injections of 0.5 µL were used to achieve sharp peaks and minimal dispersion.

Použitá instrumentace

• Shimadzu Nexera X2 or equivalent UHPLC system
• Shim-pack GIST Amide column, 150 mm × 2.1 mm I.D., 5 µm (P/N: 227-30824-06)
• UV detector set at 254 nm
• Autosampler with 0.5 µL injection capability
• Column oven maintained at 40 °C

Hlavní výsledky a diskuse

The method achieved baseline separation of all eight analytes—caffeine, theophylline, theobromine, hypoxanthine, adenine, xanthine, guanine and uric acid—within a short analysis time. Chromatograms displayed sharp, symmetric peaks with retention times reflecting increasing polarity. Reproducibility studies indicated relative standard deviations below 1.5% for retention times and peak areas.

Přínosy a praktické využití metody

• Rapid profiling of purine alkaloids in food and beverage samples
• Quality control in pharmaceutical formulations containing purine-based drugs
• Routine monitoring of biomarkers in biological fluids

Budoucí trendy a možnosti využití

Advances in column technology and coupling with mass spectrometry can expand the method to more complex matrices and trace-level detection. Miniaturized HILIC systems and high-throughput platforms will further enhance analysis speed and reduce solvent consumption.

Závěr

The Shim-pack GIST Amide column coupled with HILIC provides a robust, reproducible and efficient solution for separating a range of purine bases and alkaloids. The isocratic method offers simplicity and consistency, making it well-suited for quality control and research applications.

Reference

• Shimadzu Corporation. Analysis of Purine Bases on Shim-pack GIST Amide. Application Note ERAS-1000-0247, First Edition (Dec. 2021).