Quantification of Caffeine in Energy and Cola Drinks via Rapid High Performance Liquid Chromatography Assays with Ultra Violet Diode Array Detection

There is currently a lack of regulation of the caffeine found in cola and energy drinks by the FDA, which fails to protect the consumers of these products. Due to this lack of regulation, cola and energy drinks can have noticeable differences in their caffeine content when compared to the average am...

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Main Authors: Christopher E. Karlsen, Jake A. Cravino, Arianne Soliven, Peter J. Mahon, Feng Li, R. Andrew Shalliker
Format: Article
Language:English
Published: MDPI AG 2025-03-01
Series:Beverages
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Online Access:https://www.mdpi.com/2306-5710/11/2/39
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author Christopher E. Karlsen
Jake A. Cravino
Arianne Soliven
Peter J. Mahon
Feng Li
R. Andrew Shalliker
author_facet Christopher E. Karlsen
Jake A. Cravino
Arianne Soliven
Peter J. Mahon
Feng Li
R. Andrew Shalliker
author_sort Christopher E. Karlsen
collection DOAJ
description There is currently a lack of regulation of the caffeine found in cola and energy drinks by the FDA, which fails to protect the consumers of these products. Due to this lack of regulation, cola and energy drinks can have noticeable differences in their caffeine content when compared to the average amount per serving labelled on the product. In this study, we demonstrate the ability to analyse caffeine rapidly in under 20 s, and with HPLC pressures under 3500 psi (241 bar). To facilitate a high-throughput routine HPLC analysis of the caffeine content found in energy and cola drinks, two HPLC column technologies are studied, a conventional run HPLC column, and a newly commercialised Radial Flow Splitting end fitted HPLC column. The Radial Flow Splitting fitted column demonstrated the following benefits: a 37% reduction in pressure, an increased signal intensity sensitivity of 35%, a reduced analysis time by 20%, and improved metrics in assay precision based on triplicate injections associated with retention time, peak area, and peak height precision %RSD values. Both rapid HPLC methods offer greater opportunity for expanded beverage testing, which can ultimately help protect the consumer. The quantified energy drinks that were tested had a higher caffeine content, on average, than the labelled caffeine content, with an approximately ±16 mg difference per serving size for the energy drinks. In the case of the cola drinks, which did not include caffeine levels on the food label, we compared the levels to the USDA guidance and found up to double the recommended amount of caffeine in one serving for the samples studied. This highlights the need to have stricter regulations for caffeinated beverages to protect consumers and provide transparency regarding the caffeine content.
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spelling doaj-art-bbf3802e665d428f82f55db846f3546c2025-08-20T02:28:27ZengMDPI AGBeverages2306-57102025-03-011123910.3390/beverages11020039Quantification of Caffeine in Energy and Cola Drinks via Rapid High Performance Liquid Chromatography Assays with Ultra Violet Diode Array DetectionChristopher E. Karlsen0Jake A. Cravino1Arianne Soliven2Peter J. Mahon3Feng Li4R. Andrew Shalliker5School of Science, Western Sydney University, South Paramatta Campus, Sydney, NSW 2150, AustraliaSchool of Science, Western Sydney University, South Paramatta Campus, Sydney, NSW 2150, AustraliaSchool of Science, Western Sydney University, South Paramatta Campus, Sydney, NSW 2150, AustraliaSchool of Science, Computing and Engineering Technologies, Swinburne University of Technology, Hawthorn, VIC 3122, AustraliaSchool of Science, Western Sydney University, South Paramatta Campus, Sydney, NSW 2150, AustraliaSchool of Science, Western Sydney University, South Paramatta Campus, Sydney, NSW 2150, AustraliaThere is currently a lack of regulation of the caffeine found in cola and energy drinks by the FDA, which fails to protect the consumers of these products. Due to this lack of regulation, cola and energy drinks can have noticeable differences in their caffeine content when compared to the average amount per serving labelled on the product. In this study, we demonstrate the ability to analyse caffeine rapidly in under 20 s, and with HPLC pressures under 3500 psi (241 bar). To facilitate a high-throughput routine HPLC analysis of the caffeine content found in energy and cola drinks, two HPLC column technologies are studied, a conventional run HPLC column, and a newly commercialised Radial Flow Splitting end fitted HPLC column. The Radial Flow Splitting fitted column demonstrated the following benefits: a 37% reduction in pressure, an increased signal intensity sensitivity of 35%, a reduced analysis time by 20%, and improved metrics in assay precision based on triplicate injections associated with retention time, peak area, and peak height precision %RSD values. Both rapid HPLC methods offer greater opportunity for expanded beverage testing, which can ultimately help protect the consumer. The quantified energy drinks that were tested had a higher caffeine content, on average, than the labelled caffeine content, with an approximately ±16 mg difference per serving size for the energy drinks. In the case of the cola drinks, which did not include caffeine levels on the food label, we compared the levels to the USDA guidance and found up to double the recommended amount of caffeine in one serving for the samples studied. This highlights the need to have stricter regulations for caffeinated beverages to protect consumers and provide transparency regarding the caffeine content.https://www.mdpi.com/2306-5710/11/2/39high throughputHPLCradial flow splittingcaffeineenergy drinkcola
spellingShingle Christopher E. Karlsen
Jake A. Cravino
Arianne Soliven
Peter J. Mahon
Feng Li
R. Andrew Shalliker
Quantification of Caffeine in Energy and Cola Drinks via Rapid High Performance Liquid Chromatography Assays with Ultra Violet Diode Array Detection
Beverages
high throughput
HPLC
radial flow splitting
caffeine
energy drink
cola
title Quantification of Caffeine in Energy and Cola Drinks via Rapid High Performance Liquid Chromatography Assays with Ultra Violet Diode Array Detection
title_full Quantification of Caffeine in Energy and Cola Drinks via Rapid High Performance Liquid Chromatography Assays with Ultra Violet Diode Array Detection
title_fullStr Quantification of Caffeine in Energy and Cola Drinks via Rapid High Performance Liquid Chromatography Assays with Ultra Violet Diode Array Detection
title_full_unstemmed Quantification of Caffeine in Energy and Cola Drinks via Rapid High Performance Liquid Chromatography Assays with Ultra Violet Diode Array Detection
title_short Quantification of Caffeine in Energy and Cola Drinks via Rapid High Performance Liquid Chromatography Assays with Ultra Violet Diode Array Detection
title_sort quantification of caffeine in energy and cola drinks via rapid high performance liquid chromatography assays with ultra violet diode array detection
topic high throughput
HPLC
radial flow splitting
caffeine
energy drink
cola
url https://www.mdpi.com/2306-5710/11/2/39
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