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Updated: Sep 22, 2025

Caffeine Extraction, Enzymatic Activity and Gene Expression of Caffeine Synthase from Plant Cell Suspensions
Published on: October 2, 2018
A Green High-Performance Thin-Layer Chromatography Method for the Determination of Caffeine in Commercial Energy
Ahmed I Foudah1, Faiyaz Shakeel2, Mohammad A Salkini1
1Department of Pharmacognosy, College of Pharmacy, Prince Sattam Bin Abdulaziz University, Al-Kharj 11942, Saudi Arabia.
Abstract:
The literature on green analytical approaches for caffeine estimation is limited. As a consequence, this study aimed to establish a reverse-phase high-performance thin-layer chromatography (HPTLC) technique for caffeine estimation in a variety of commercial energy drinks (ED) and pharmaceutical formulations that is rapid, sensitive, and green. The combination of ethanol-water (55:45 v v−1) was used as a mobile phase. The detection of caffeine was carried out at 275 nm. The green reverse-phase HPTLC method was linear in the concentration range of 50−800 ng band−1. Furthermore, the developed method for caffeine estimation was simple, quick, economical, accurate, precise, robust, sensitive, and green. The amount of caffeine in different marketed ED (ED1−ED10) was recorded in the range of 21.02−37.52 mg 100 mL−1 using the developed HPTLC method. However, the amount of caffeine in different commercial formulations (F1−F3) was estimated as 10.63−20.30 mg 100 mL−1 using the same method. The “analytical GREEnness (AGREE)” scale for the developed analytical method was predicted to be 0.80, utilizing 12 distinct components of green analytical chemistry, indicating the HPTLC approach’s excellent greener profile. Overall, the developed method for estimating caffeine in marketed ED and dosage forms was found to be reliable.
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