开发和验证超高效液相色谱 (UPLC) 法用于非酒精软饮料和能量饮料中咖啡因的定量测定。

Mohammad Al-Mana College for Medical Sciences, Department of Pharmacy, Dammam 34222, Saudi Arabia.

College of Pharmacy, Taif University, Department of Clinical Pharmacy, P.O. Box 11099, Taif 21944 Saudi Arabia.

J AOAC Int. 2022 Jun 29;105(4):1146-1152. doi: 10.1093/jaoacint/qsac016.

BACKGROUND

The popularity of beverages such as cold coffee, iced tea, and energy drinks has risen tremendously among athletes and youths. Energy drinks are used to enhance performance due to the presence of a high amount of caffeine (CFN) and sugars, as well as other constituents such as vitamins, amino acids, taurine, extracts of Ginkgo biloba, ginseng, guarana, and other herbal products. Commercial drinks are promoted as being beneficial to health; thus it is an important concern regarding adverse effects linked with these drinks or products.

OBJECTIVE

The aim of the study is to develop and validate the ultra performance liquid chromatography-photodiode array detector (UPLC-PDA) method for the estimation of CFN in eight marketed non-alcoholic drinks including two soft drinks and six energy drinks.

METHOD

The chromatographic separation of CFN was achieved on Acquity UPLC® CSHTM C18 1.7 µm, 2.1 × 100 mm column, using isocratic mode, mobile phase comprising acetonitrile and water (30:70, v/v) at a flow rate 0.3 mL/min using injection volume 1 µL.

RESULTS

The elution of CFN occurred at 1.06 min, and the calibration curve of the CFN was computed from the peak area ratio detected at 273 nm. All the validation parameters were found within the assay variability limits as per ICH guidelines. The obtained results revealed that the soft drinks SD1 and SD2 have 87.21 ± 1.28 and 101.81 ± 1.52% (w/w), whereas CFN concentration in energy drink brands ED1, ED2, ED3, ED4, ED5, and ED6 was 95.90 ± 1.62, 64.0 ± 1.07, 76.68 ± 1.95, 74.97 ± 2.33, 82.09 ± 2.43, and 88.04 ± 2.94% (w/w) of labeled claims, respectively.

CONCLUSIONS

The developed UPLC method was found suitable for the quality control of commercial soft and energy drinks containing CFN.

HIGHLIGHTS

The developed chromatographic method is very simple, cost effective and could be utilized for the routine analysis of caffeine in the soft and energy drinks.

背景

冷咖啡、冰茶和能量饮料等饮品在运动员和年轻人中非常受欢迎。由于含有大量咖啡因 (CFN) 和糖以及其他成分，如维生素、氨基酸、牛磺酸、银杏叶、人参、瓜拉那和其他草药提取物，能量饮料被用于提高表现。商业饮料被宣传为对健康有益；因此，与这些饮料或产品相关的不良影响是一个重要的关注点。

目的

本研究旨在开发和验证超高效液相色谱-光电二极管阵列检测器 (UPLC-PDA) 方法，用于测定两种软饮料和六种能量饮料在内的八种市售非酒精饮料中的 CFN 含量。

方法

在 Acquity UPLC® CSHTM C18 1.7μm，2.1×100mm 柱上，采用等度模式，流动相为乙腈和水（30:70，v/v），流速为 0.3mL/min，进样量为 1μL，实现 CFN 的色谱分离。

结果

CFN 在 1.06min 洗脱，CFN 的校准曲线是根据 273nm 处检测到的峰面积比计算得出的。根据 ICH 指南，所有验证参数均在分析变异性限度内。所得结果表明，软饮料 SD1 和 SD2 的 CFN 浓度分别为 87.21±1.28%（w/w）和 101.81±1.52%（w/w），而能量饮料品牌 ED1、ED2、ED3、ED4、ED5 和 ED6 的 CFN 浓度分别为 95.90±1.62%（w/w）、64.0±1.07%（w/w）、76.68±1.95%（w/w）、74.97±2.33%（w/w）、82.09±2.43%（w/w）和 88.04±2.94%（w/w），分别与标签声称值相符。