A Fast Determination of Chlorophylls in Barley Grass Juice Powder Using HPLC Fused-Core Column Technology and HPTLC
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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 performance 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 separated 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 comparative 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 techniques for the characterisation of barley juice extracts in quality control.
KeywordsBarley Dietary supplement Chlorophyll HPLC HPTLC
The authors gratefully acknowledge the financial support of the Charles University in Prague, project UNCE no. 204026/2012.
Conflict of Interest
Lucie Havlíková declares that she has no conflict of interest. Dalibor Šatínský declares that he has no conflict of interest. Lubomír Opletal declares that he has no conflict of interest. Petr Solich declares that he has no conflict of interest. This article does not contain any studies with human or animal subjects.
- Bhat SR (2005) Science Reporter 42:29Google Scholar
- EDQM (2010) European pharmacopoeia, 7th Edn. EDQM, Strasbourg, pp 70Google Scholar
- ICH Expert Working Group (2005) ICH guidelines. http://www.ich.org/fileadmin/Public_Web_Site/ICH_Products/Guidelines/Quality/Q2_R1/Step4/Q2_R1__Guideline.pdf (Accessed 08 March, 2013)
- Kao TH, Chen ChJ, Chen BH (2011) Talanta 86:349Google Scholar
- Lee SH, Jew SS, Chang PS, Hong IJ, Hwang ES, Kim KS, Kim KT (2003) Food Sci Biotechnol 12(3):268Google Scholar
- Leung AY (2010) J AOAC Int 93(5):1355Google Scholar
- Paulíčková I, Ehrenbergerová J, Fiedlerová V et al (2007) Czech J Food Sci 25:65Google Scholar
- Sigma-Aldrich (2008) http://www.lsc.gr/attachments/File/ASCENTIS_EXPRESS.pdf, (Accessed 08 March, 2013)
- Way WK, Santasania CT (2012) Ascentis® Express 5 micron: a fused-core particle HPLC column for faster HPLC separations with no backpressure concerns. The application notebook LC/GC 06, 54. Supelco/Sigma-Aldrich, St. LouisGoogle Scholar