Food Analytical Methods

, Volume 7, Issue 3, pp 629–635 | Cite as

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


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.


Barley 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.


  1. AbuMweis SS, Jew S, Ames NP (2010) Eur J Clin Nutr 64(12):1472CrossRefGoogle Scholar
  2. Bhat SR (2005) Science Reporter 42:29Google Scholar
  3. Bohn T, Walczyk T (2004) J Chrom A 1024(1–2):123CrossRefGoogle Scholar
  4. Burns J, Fraser PD, Bramley PM (2003) Phytochemistry 62(6):939CrossRefGoogle Scholar
  5. Canjura FL, Schwartz SJ (1991) J Agric Food Chem 39(6):1102CrossRefGoogle Scholar
  6. Cano MP (1991) J Agric Food Chem 39(10):1786CrossRefGoogle Scholar
  7. Choi JS, Kim H, Jung MH, Hong S, Song J (2009) Mol Nutr Food Res 54(7):1004CrossRefGoogle Scholar
  8. EDQM (2010) European pharmacopoeia, 7th Edn. EDQM, Strasbourg, pp 70Google Scholar
  9. Ferruzzi MG, Böhm V, Courtney PD, Schwartz SJ (2002) J Food Sci 67(7):2589CrossRefGoogle Scholar
  10. Fu PP, Chiang HM, Xia Q, Chen T, Chen BH, Yin JJ, Wen KC, Lin G, Yu H (2009) J Environ Sci Health C Environ Carcinog Ecotoxicol Rev 27(2):91–119CrossRefGoogle Scholar
  11. ICH Expert Working Group (2005) ICH guidelines. (Accessed 08 March, 2013)
  12. Izydorczyk MS, Dexter JE (2008) Food Res Int 41(9):850CrossRefGoogle Scholar
  13. Janda T, Szalai G, Rios-Gonzalez K, Veisz O, Páldi E (2003) Plant Sci 164(2):301CrossRefGoogle Scholar
  14. Kao TH, Chen ChJ, Chen BH (2011) Talanta 86:349Google Scholar
  15. Lanfer-Marquez UM, Barros RMC, Sinnecker P (2005) Food Res Int 38:885CrossRefGoogle Scholar
  16. Lee SH, Jew SS, Chang PS, Hong IJ, Hwang ES, Kim KS, Kim KT (2003) Food Sci Biotechnol 12(3):268Google Scholar
  17. Leung AY (2010) J AOAC Int 93(5):1355Google Scholar
  18. Madhujith T, Izydorczyk MS, Shahidi F (2006) Agr Food Chem 54(9):3283CrossRefGoogle Scholar
  19. Mangos TJ, Berger RG (1997) Z Lebensm Unters Forsch A 204:345CrossRefGoogle Scholar
  20. Mateos R, García-Mesa JA (2006) Anal Bioanal Chem 385(7):1247CrossRefGoogle Scholar
  21. Nakajima S, Hagiwara Y, Hagiwara H, Shibamoto T (1998) J Agric Food Chem 46:1529CrossRefGoogle Scholar
  22. Nakamura A, Watanabe T (1998) FEBS Lett 426(2):201CrossRefGoogle Scholar
  23. Paulíčková I, Ehrenbergerová J, Fiedlerová V et al (2007) Czech J Food Sci 25:65Google Scholar
  24. Schoefs B (2002) Trends Food Sci Tech 13(11):361CrossRefGoogle Scholar
  25. Schütz K, Muks E, Carle R, Schieber A (2006) J Agric Food Chem 54(23):8812CrossRefGoogle Scholar
  26. Sigma-Aldrich (2008), (Accessed 08 March, 2013)
  27. Sullivan D, Crowley R (2006) Toxicology 221(1):28CrossRefGoogle Scholar
  28. 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

Copyright information

© Springer Science+Business Media New York 2013

Authors and Affiliations

  • Lucie Havlíková
    • 1
  • Dalibor Šatínský
    • 1
  • Lubomír Opletal
    • 1
  • Petr Solich
    • 1
  1. 1.Faculty of Pharmacy, Department of Analytical ChemistryCharles University in PragueHradec KrálovéCzech Republic

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