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Updated: Oct 21, 2025

Caffeine Extraction, Enzymatic Activity and Gene Expression of Caffeine Synthase from Plant Cell Suspensions
Published on: October 2, 2018
[Determination of Caffeine, Theobromine, and Theophylline in Chocolate Using LC-MS]
Tomoya Tanaka1, Keisuke Kimura1, Kimiko Kan1
1Tokyo Metropolitan Institute of Public Health.
Abstract:
A simultaneous determination method for caffeine, theobromine, and theophylline in chocolate was developed. Three compounds were ultrasonically extracted twice (15 min at 50℃) in an acetonitrile-water (1 : 1, v/v). The extract was purified using Oasis HLB SPE cartridge, and the purified processed by LC-MS. The method exhibited recoveries of 97.4-100.2%, RSDs of repeatability of 1.0-2.8%, and RSDs of within-laboratory reproducibility of 2.0-7.9%. This method was simpler and more selective than existing methods, and was practical for caffeine, theobromine, and theophylline analysis in chocolate.
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