Simultaneous Determination of Caffeine and Water-Soluble Vitamins in Energy Drinks by HPLC with Photodiode Array and Fluorescence Detection

Abstract

The high-performance liquid chromatography (HPLC) method with photodiode array (PDA) and fluorescence (FL) detection for the simultaneous separation and determination of caffeine and water-soluble vitamins is described. The method is relatively rapid: 13 compounds can be analyzed within 30 min: caffeine; ascorbic acid (vitamin C); thiamine (vitamin B 1 ); riboflavin 5-phosphate (FMN, flavin mononucleotide) and riboflavin (vitamin B 2 ); nicotinic acid and nicotinamide (vitamin B 3 ); pantothenic acid (vitamin B 5 ); pyridoxal, pyridoxamine, and pyridoxine (vitamin B 6 ); folic acid (vitamin B 9 ); and cyanocobalamin (vitamin B 12 ). This HPLC method is designated for quantification of caffeine and water-soluble vitamins in liquid and tablet energy drinks as well as for determination of vitamins in vitamin supplements. The method was validated in terms of linearity, limit of detection (LOD) and quantification (LOQ), accuracy, and instrument precision. The LODs determined using HPLC-PDA ranged from 13 to 121 ng mL −1 and using HPLC-FL were between 8 and 61 ng mL −1. Intra-and interday instrument precisions for liquid energy drink, expressed as relative standard deviation (RSD), were less than 1.87 and 1.73 %, respectively, and less than 1.44 and 1.83 %, respectively, for tablet energy drink. The method could be useful in the quantitative analysis of caffeine and of water-soluble vitamins in energy drinks and vitamin supplements.

Gluten-Free Flours from Different Raw Materials as the Source of Vitamin B₁, B₂, B₃ and B₆

“…In Figs. 1-3, the separation of vitamins B2, B3, and B6 obtained using the HPLC method employed in the present study (30) is presented. Figure 1 shows the HPLC chromatogram of standards of vitamin B2, B3, and B6.…”

“…Two chromatographic methods were applied for all extracts-one for B1 vitamin (thiamine) (29) and another for B2 (riboflavin), B3 (as nicotinamide), and B6 vitamin (as the sum of pyridoxine, pyridoxamine, and pyridoxal) (30). The conditions for both methods are summarized in Table 2.…”

Development of ic-ELISA and lateral-flow immunochromatographic strip for detection of vitamin B2 in an energy drink and vitamin tablets

“…Currently, the available methods to determine vitamin B 2 include microbiological assays (Barton-Wright & Booth, 1943;Hyma, 1945), high-performance liquid chromatography (HPLC); (Hampel, York, & Allen, 2012;Marti-Andres, Escuder-Gilabert, MartinBiosca, Sagrado, & Medina-Hernandez, 2015;Schmidt, Schreiner, & Mayer, 2017), electrochemical methods (Kowalczyk, Sadowska, Krasnodebska-Ostrega, & Nowicka, 2017), and fluorescence spectroscopy (Chen, Li, Cui, Yu, & Zhai, 2015;Gliszczyńska-Świglo & Rybicka, 2015;Privitera & Lozano, 2017). Microbiological assays, which are based on Lactobacillus rhamnosus, are highly sensitive.…”

Analytical Method Validation and Determination of Pyridoxine, Nicotinamide, and Caffeine in Energy Drinks Using Thin Layer Chromatography-Densitometry

“…Determination of caffeine in energy drinks were performed by HPLC/UV [7][8][9], HPLC/PDA and fluorescence detection [10], microemulsion electrokinetic chromatography [11], mass spectrometry [12], and spectrophotometry UV [13,19]. Determination of vitamin B in energy drinks and multivitamins were performed by HPLC/UV methods [14][15][16][17], HPLC with DAD, ELSD, and MS detection [18], and spectrophotometry UV [19].…”