A Fast Determination of Chlorophylls in Barley Grass Juice Powder Using HPLC Fused-Core Column Technology and HPTLC Lucie Havlíková & Dalibor Šatínský & Lubomír Opletal & Petr Solich Received: 18 March 2013 / Accepted: 12 June 2013 / Published online: 22 June 2013 # Springer Science+Business Media New York 2013 Abstract This work presents a novel approach to qualitative and quantitative evaluation of chlorophyll content in food supplement using simple and fast separation techniques (high performance liquid chromatography (HPLC) and high perfor- mance thin layer chromatography (HPTLC)). For the first time, an Ascentis® Express C-18 HPLC column (100×4.6 mm; 2.7 μm), based on fused-core particles, was utilised for the analysis of chlorophyll a and chlorophyll b in green young barley grass juice powder extracts. The mobile phase was composed of methanol, water and ethyl acetate of 40:10:50 (v/v/v) at a flow rate of 1.5 mL/min. Chlorophylls were sepa- rated in time less than 6 min. The extraction procedure was optimised by using of 90 % of acetone in water, and the method was validated. The method precision was found in the range of 1.15–2.85 %, and the method recovery was established at three concentration levels. Calibration graphs were linear in the range of 0.75–25.5 mg/L, with correlation coefficients r 2 =0.99958 for chlorophyll a and r 2 =0.99961 for chlorophyll b. The com- parative HPTLC fingerprint analysis of three samples from different continents showed only slight qualitative differences between the particular samples. The results demonstrated that the proposed methods can be used as fast and powerful tech- niques for the characterisation of barley juice extracts in quality control. Keywords Barley . Dietary supplement . Chlorophyll . HPLC . HPTLC Introduction The consumption of dietary supplements of plant origin and plant-based foods has rapidly increased worldwide. There are two reasons for the growing consumption of such supplements—advertisements of their beneficial health effects and the strengthening of disease preven- tion among populations. The tremendous popularity of these products has resulted in increased scrutiny from consumers, health professionals and regulators about the quality and levels of active ingredients in these products (Sullivan and Crowley 2006). The boom in the use of botanical ingredients and nutraceutical products is per- haps one of the most challenging tasks that the analyt- ical community must face. To ensure consumer health protection, the quality and safety of herbal plants, par- ticularly those used for dietary supplement preparations, must be determined (Fu et al. 2009). As herbal supple- ments are neither food nor medicine, they often inhabit a grey area between the two, which makes legal control extremely difficult (Schütz et al. 2006). Plant dietary supplements are often complex matrices that require a specific procedure to extract and analyse required in- gredients. Separation methods (high performance liquid chromatography (HPLC), HPLC tandem mass spectrom- etry, thin layer chromatography (TLC)) are among the most widely used techniques to analyse such complex matrices. Young barley (Hordeum vulgare) contained in dietary supplements is mainly produced in the form of juice. The content of the active ingredients depends on the region of origin, the average rainfall and the harvest technique and time (Paulíčková et al. 2007). Young barley is harvested when it is approximately 30 cm tall and is juiced before being turned into powder. This step is important to break L. Havlíková (*) : D. Šatínský : L. Opletal : P. Solich Faculty of Pharmacy, Department of Analytical Chemistry, Charles University in Prague, Heyrovského 1203, 500 12 Hradec Králové, Czech Republic e-mail: lucie.havlikova@faf.cuni.cz Food Anal. Methods (2014) 7:629–635 DOI 10.1007/s12161-013-9665-x