Development and validation of an RP-HPLC method for the determination of caffeine and preservatives (sodium benzoate and potassium sorbate) in sports and energy drinks
Mirjana S. Jankulovska, Simona Stojchevska, Lenche Velkoska-Markovska
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引用次数: 0
Abstract
Preservatives in sports and energy drinks play a crucial role, especially in maintaining their quality and safety, while caffeine is included mainly in energy drinks as a stimulant to enhance alertness, focus, and physical performance. In this study, a rapid, simple and sensitive RP-HPLC method for simultaneous determination of caffeine, sodium benzoate (SB) and potassium sorbate (PS) in sports and energy drinks was developed. Separation of analytes was achieved using Poroshell 120 EC - C18 (3.0 × 50 mm, 2.7 µm) analytical column. The best results were obtained by isocratic elution with mobile phase consisting of acetonitrile and phosphate buffer (KH2PO4/H3PO4) with pH = 3.8 in volume ratio 85/15 (V/V), flow rate of 1.0 mL/min, and a constant column temperature of (25 °C). The detection was performed at 230, 260 and 275 nm. The developed RP-HPLC method was validated regarding the selectivity/specificity, linearity (R2 ≥ 0.996), accuracy (recoveries between 95.99 and 101.5%), precision (intra-day RSD < 1%), limit of detection (LOD) and limit of quantification (LOQ). All the validation parameters were within the acceptance range. The linearity of the RP-HPLC method was confirmed with R2 values higher than 0.996 in the concentration range of 49.95 - 449.55 mg/L for caffeine, 29.97 - 269.73 mg/L for SB, and 60.53 - 544.85 mg/L for PS. Intra-day repeatability data demonstrated that proposed method was reliable, and it is characterized by high precision (RSD \(\le\) 0.49%). The recovery data ranged between 95.99 - 101.50%, indicating excellent recoveries for SB, PS and caffeine by the proposed RP-HPLC method. LODs were 0.015 mg/L for caffeine, 0.025 mg/L for SB, and 0.018 mg/L for PS, while LOQs were 0.083 mg/L for SB, 0.059 mg/L for PS and 0.049 mg/L for caffeine. The proposed method was successfully applied for determination of caffeine, SB and PS in sports and energy drinks.
运动饮料和能量饮料中的防腐剂起着至关重要的作用，尤其是在保持其质量和安全方面，而能量饮料中的咖啡因主要是作为一种兴奋剂来提高警觉性、注意力和身体表现。建立了一种快速、简便、灵敏的同时测定运动饮料和功能饮料中咖啡因、苯甲酸钠（SB）和山梨酸钾（PS）含量的反相高效液相色谱方法。采用Poroshell 120 EC - C18 （3.0 × 50 mm, 2.7µm）分析柱进行分离。流动相为乙腈-磷酸盐缓冲液（KH2PO4/H3PO4）， pH = 3.8，体积比为85/15 (V/V)，流速为1.0 mL/min，柱温为（25℃），等容洗脱效果最佳。在230、260和275 nm处进行检测。方法的选择性/特异性、线性度（R2≥0.996）、准确度（加样回收率在95.99 ～ 101.5之间）良好%), precision (intra-day RSD < 1%), limit of detection (LOD) and limit of quantification (LOQ). All the validation parameters were within the acceptance range. The linearity of the RP-HPLC method was confirmed with R2 values higher than 0.996 in the concentration range of 49.95 - 449.55 mg/L for caffeine, 29.97 - 269.73 mg/L for SB, and 60.53 - 544.85 mg/L for PS. Intra-day repeatability data demonstrated that proposed method was reliable, and it is characterized by high precision (RSD \(\le\) 0.49%). The recovery data ranged between 95.99 - 101.50%, indicating excellent recoveries for SB, PS and caffeine by the proposed RP-HPLC method. LODs were 0.015 mg/L for caffeine, 0.025 mg/L for SB, and 0.018 mg/L for PS, while LOQs were 0.083 mg/L for SB, 0.059 mg/L for PS and 0.049 mg/L for caffeine. The proposed method was successfully applied for determination of caffeine, SB and PS in sports and energy drinks.
期刊介绍：
This interdisciplinary journal publishes new measurement results, characteristic properties, differentiating patterns, measurement methods and procedures for such purposes as food process innovation, product development, quality control, and safety assurance.
The journal encompasses all topics related to food property measurement and characterization, including all types of measured properties of food and food materials, features and patterns, measurement principles and techniques, development and evaluation of technologies, novel uses and applications, and industrial implementation of systems and procedures.
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