HPLC Method for Analysis of Caffeine, Theobromine, Theophylline  on Primesep SB

Caffeine is a natural stimulant and methylxanthine alkaloid. with the molecular formula C₆H₁₀N₄O₂. Caffeine can be found in a variety of plants, including tea, coffee, cocoa, kola nuts, and guarana. Ingestion of it can increase alertness and cognitive function. It can also cause worsening anxiety, heart palpitations, and headaches.

Theophylline is a methylxanthine compound with the molecular formula C₇H₈N₄O₂. It is a caffeine metabolite and is often used in forensic analyses. Besides analytical use, it is often used in treating respiratory conditions.

Theobromine is a methylxanthine alkaloid with the molecular formula C₇H₈N₄O₂. It is a derivative of caffeine. It can be found in many foods, including chocolate and tea. Despite that, it is toxic to dogs.

Caffeine, Theobromine, Theophylline can be retained and analyzed using the Primesep SB stationary phase column. The analysis utilizes an isocratic method with a simple mobile phase consisting of water and ethanol (EtOH) with a phosphoric acid buffer. Detection is performed using UV.