Skip to main content
SpringerLink
Log in

Determination of Theanine and Catechin in Camellia sinensis (Kangra Tea) Leaves by HPTLC and NMR Techniques

  • Published:
Food Analytical Methods Aims and scope Submit manuscript

Abstract

Camellia sinensis is used to make special tea and also become a raw material for dietary supplements, health foods, and cosmeceuticals. Theanine (amino acid) and catechins (polyphenol) are most abundantly found in tea leaves. Theanine and catechins are also responsible for taste and flavor of tea products along with therapeutic and nutritional values. Hence, identification and quantification method for theanine and catechin has been developed using high performance thin layer chromatography (HPTLC) and NMR techniques. HPTLC method was validated for selectivity, sensitivity, accuracy, and precision. Theanine and catechin were quantified in three tea samples including randomly plucked tea leaves (HAR), authentically plucked tea leaves (HAN), and processed tea (HAT). Both theanine and catechin were found maximum in HAN as compared to HAR and HAT. The Validated HPTLC method is economical, rapid, and highly suitable for quantification of theanine and catechin. Moreover NMR studies were also used for the qualitative and quantitative estimation of theanine and catechin along with other important metabolites in C. sinensis samples.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3

Similar content being viewed by others

Abbreviations

HPTLC:

high performance thin layer chromatography

LOD:

limit of detection

LOQ:

limit of quantification

NP-HPTLC:

normal phase-high performance thin layer chromatography

HAR:

randomly plucked leaves from the tea farm

HAN:

fresh leaves plucked by authentic method from the farm

HAT:

processed tea leaves

TH:

theanine

CH:

catechin

References

  • Alam P, Ali M, Singh R, Shakeel F (2011) A new HPTLC densitometric method for analysis of swertiamarin in Enicostemma littorale and commercial formulations. Nat Prod Res 25(1):17–25

    Article  CAS  Google Scholar 

  • Bhatt PR, Pandya KB, Sheth NR (2010) Camellia sinensis (L): the medicinal beverage: a review. Int J Pharm Sci Rev Res 3(2):1–9

    Google Scholar 

  • Casimir J, Jadot J, Renard M (1960) Separation and characterization of N-ethyl-γ-glutamine from Xerocomus badius. Biochim Biophys Acta Mol Cell Biol Lipids 39:462–468

    Article  CAS  Google Scholar 

  • Chen C, Liang C, Lai J, Tsai Y, Tsay J, Lin J (2003) Capillary electrophoretic determination of theanine, caffeine, and catechins in fresh tea leaves and oolong tea and their effects on rat neurosphere adhesion and migration. J Agric Food Chem 51:7495–7503

    Article  CAS  Google Scholar 

  • Chu D, Kobayashi K, Juneja LR, Yamamoto T (1997) Theanine—its synthesis, isolation, and physiological activity. In: Yamamoto T, Juneja LR, Chu D, Kim M (eds) Chemistry and applications of green Tea. CRC Press, Boca Raton, pp 129–135

    Google Scholar 

  • Chu D, Okubo T, Nagato Y, Yokogoshi H (1999) L-theanine—a unique amino acid of green tea and its relaxation effect in humans. Trends Food Sci Technol 10:199–204

    Article  Google Scholar 

  • Crozier A, Yokota T, Jaganath IB, Marks SC, Saltmarsh M, Clifford MN (2006) Secondary metabolites in fruits, vegetables, beverages and other plant-based dietary components. In: Crozier A, Clifford MN, Ashihara H (eds) Plant secondary metabolites—occurrence, structure and role in the human diet. Blackwell, Oxford, pp 208–302

    Google Scholar 

  • Eschenauer G, Sweet BV (2006) Pharmacology and therapeutic uses of theanine. Am J Health Syst Pharm 63(1):26–30

    Article  CAS  Google Scholar 

  • Fan G, Zhang MY, Zhou XD, Lai XR, Yue QH, Tang C, Luo WZ, Zhang Y (2012) Quality evaluation and species differentiation of Rhizoma coptidis by using proton nuclear magnetic resonance spectroscopy. Anal Chim Acta 747:76–83

    Article  CAS  Google Scholar 

  • Fan G, Luo WZ, Luo SH, Li Y, Meng XL, Zhou XD, Zhang Y (2014) Metabolic discrimination of Swertia mussotii and Swertia chirayita known as “Zangyinchen” in traditional Tibetan medicine by 1H NMR-based metabolomics. J Pharm Biomed Anal 98:364–370

    Article  CAS  Google Scholar 

  • Gramza A, Korczak J, Amarowicz R (2005) Tea polyphenols—their antioxidant properties and biological activity—a review. Pol J Food Nutr Sci 14/55(3):219–235

    Google Scholar 

  • Hampton MG (1992) Production of black tea. In: Willson KC, Clifford MN (eds) Tea: CultiVation to consumption. Chapman and Hall, London, p 555

    Google Scholar 

  • Isemura M, Saeki K, Kimura T, Hayakawa S, Minami T, Sazuka M (2000) Tea catechins and related polyphenols as anti-cancer agents. BioFactors 13:81–85

    Article  CAS  Google Scholar 

  • Kakuda T (2002) Neuroprotective effects of the green tea components theanine and catechins. Biol Pharm Bull 25(12):1513–1518

    Article  CAS  Google Scholar 

  • Kim HK, Choi YH, Verpoorte R (2010) NMR-based metabolomic analysis of plants. Nat Protoc 5:536–549

    Article  CAS  Google Scholar 

  • Kito M, Kokura H, Izaki J, Sasaoka K (1968) Theanine, a precursor of the phloroglucinol nucleus of catechins in tea plants. Phytochemistry 7:599–603.9

    Article  CAS  Google Scholar 

  • Le GG, Colquhoun IJ, Defernez M (2004) Metabolite profiling using 1H NMR spectroscopy for quality assessment of green tea, Camellia sinensis (L.). J Agric Food Chem 52(4):692–700

    Article  Google Scholar 

  • Lee JE, Lee BJ, Chung JO, Hwang JA, Lee SJ, Lee CH, Hong YS (2010) Geographical and climatic dependencies of green tea (Camellia sinensis) metabolites: a (1) H NMR-based metabolomics study. J Agric Food Chem 58(19):10582–10589

    Article  CAS  Google Scholar 

  • Lee JE, Lee BJ, Hwang JA, Ko KS, Chung JO, Kim EH, Lee SJ, Hong YS (2011a) Metabolic dependence of green tea on plucking positions revisited: a metabolomic study. J Agric Food Chem 59(19):10579–10585. doi:10.1021/jf202304z

    Article  CAS  Google Scholar 

  • Lee J, Lee B, Chung J, Shin H, Lee S, Lee C, Hong Y (2011b) 1H NMR-based metabolomic characterization during green tea (Camellia sinensis) fermentation. Food Res Int 44:597–604

    Article  Google Scholar 

  • Liu Q, Duan H, Luan J, Yagasaki K, Zhang G (2009) Effects of theanine on growth of human lung cancer and leukemia cells as well as migration and invasion of human lung cancer cells. Cytotechnology 59(3):211–217

    Article  CAS  Google Scholar 

  • Nagata T, Sakai S (1984) Differences in caffeine, flavonols and amino acids contents in leaves of cultivated species of Camellia. Jpn J breed 34:459–467

    Article  CAS  Google Scholar 

  • Namita P, Mukesh R, Vijay JK (2012) Camellia sinensis (green tea): a review. Global J Pharmacol 6(2):52–59

    Google Scholar 

  • Reich E, Schibli A, Widmer V (2006) HPTLC methods for identification of green tea and green tea extract. J Liq Chromatogr Relat Technol 29:2141–2151

    Article  CAS  Google Scholar 

  • Sakato Y (1949) The chemical constituents of tea: III. A new amide theanine. Nippon Nogeikagaku Kaishi 23:262–267 (in Japanese)

    Article  Google Scholar 

  • Vuong QV, Bowyer MC, Roach PD (2011) L-Theanine: properties, synthesis and isolation from tea. J Agric Food Chem 91:1931–1939

    Article  CAS  Google Scholar 

  • Wang L, Wu D, Xue Y (2012) Taste modifying compositions. Patent WO 2012054743 A2

  • Yuan S, Song Y, Jing W, Wang Y, Yang X, Liu D (2014) Simultaneous determination of caffeine, gallic acid, theanine, (−)-epigallocatechin and (−)-epigallocatechin-3-gallate in green tea using quantitative 1H-NMR spectroscopy. Anal Methods 6:631–631

    Article  Google Scholar 

Download references

Acknowledgments

The authors are grateful to the Director of the institute for providing necessary facilities and to CSIR, New Delhi, for providing financial help under the network project MLP0066. IHBT Publication No. for this research paper is 3810.

Author information

Authors and Affiliations

  1. Natural Product Chemistry and Process Development Division, CSIR-Institute of Himalayan Bioresource Technology, Palampur, 176 061 (HP), India

    Dinesh Kumar & Upendra Sharma

  2. Hill Area Tea Science Division, CSIR-Institute of Himalayan Bioresource Technology, Palampur, 176 061 (HP), India

    Ashu Gulati

Authors
  1. Dinesh Kumar
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Ashu Gulati
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Upendra Sharma
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding authors

Correspondence to Dinesh Kumar or Upendra Sharma.

Ethics declarations

This article does not contain any studies with human participants or animals performed by any of the authors.

Informed Consent

Not applicable.

Conflict of Interest

The authors declare that they have no competing interests.

Additional information

Highlights

•Development of HPTLC-based validated method for the quantification of theanine and catechin in Camellia sinensis

•Metabolic profiling of Camellia sinensis using NMR

•Quality control method for Camellia sinensis and its products

Electronic supplementary material

Below is the link to the electronic supplementary material.

ESM 1

ᅟ

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Kumar, D., Gulati, A. & Sharma, U. Determination of Theanine and Catechin in Camellia sinensis (Kangra Tea) Leaves by HPTLC and NMR Techniques. Food Anal. Methods 9, 1666–1674 (2016). https://doi.org/10.1007/s12161-015-0343-z

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12161-015-0343-z

Keywords

Search

Navigation