Author(s):
Eka Indra Setyawan, Erna Prawita Setyowati, Abdul Rohman, Akhmad Kharis Nugroho
Email(s):
a.k.nugroho@ugm.ac.id
DOI:
10.5958/0974-360X.2020.00271.1 
Address:
Eka Indra Setyawan1,2, Erna Prawita Setyowati2, Abdul Rohman2, Akhmad Kharis Nugroho2*
1Department of Pharmacy, Faculty of Mathematics and Natural Science, Udayana University, Bali, Indonesia.
2Faculty of Pharmacy, Gadjah Mada University, Yogyakarta, Indonesia.
*Corresponding Author
Published In:
Volume - 13,
Issue - 3,
Year - 2020
ABSTRACT:
This study was aimed to develop sensitive, selective, and simple methods for determining the levels of, catechin, caffeine, and epigallocatechin gallate in green tea leaf extract. The analytical separation was performed using C-18 column and temperature was set on 15 oC. The mobile phase used was ortho-phosphoric acid: water: methanol (0.1:79.9:20 v/v/v) pH 3.1 at flow rate 1.0 ml/min. Analyte detection was performed at 280 nm. The analytes were successfully separated with retention time of epigallocatechin gallate, catechin, and caffeine were 16.189, 8.609, and 11.986 minutes, respectively. Linearity of epigallocatechin gallate, catechin, and caffeine were established in the range of 0.5-30 ppm, 0.1-0.6 ppm, and 0.5-5 ppm, respectively. The method was validated as per AOAC guidelines. The level of epigallocatechin gallate, catechin, and caffeine to be founded were 364.73 ± 0.21, 13.86 ± 0.02, and 78.60 ± 0.03 mg/g dry weight, respectively.
Cite this article:
Eka Indra Setyawan, Erna Prawita Setyowati, Abdul Rohman, Akhmad Kharis Nugroho. Simultaneous Determination of Epigallocatechin Gallate, Catechin, and Caffeine from Green Tea Leaves (Camellia sinensis L) Extract by RP-HPLC. Research J. Pharm. and Tech 2020; 13(3):1489-1494. doi: 10.5958/0974-360X.2020.00271.1
Cite(Electronic):
Eka Indra Setyawan, Erna Prawita Setyowati, Abdul Rohman, Akhmad Kharis Nugroho. Simultaneous Determination of Epigallocatechin Gallate, Catechin, and Caffeine from Green Tea Leaves (Camellia sinensis L) Extract by RP-HPLC. Research J. Pharm. and Tech 2020; 13(3):1489-1494. doi: 10.5958/0974-360X.2020.00271.1 Available on: https://rjptonline.org/AbstractView.aspx?PID=2020-13-3-76
REFERENCES:
1. Li D, Martini N, Wu Z, Wen J. Development of an isocratic HPLC method for catechin quantification and its application to formulation studies. Fitoterapia. 2012;83(7):1267–1274.
2. Beak S-G, Kim D-J. Effects of Green Tea intake and Flexible Exercise on Oxidative Stress and Catecholamines. Res. J. Pharm. Technol. 2017;10(11):3881.
3. Martono Y, Martono S. High-performance liquid chromatography analysis for determination of gallic acid, caffeine, and epigallocatechin gallate concentration in various tea bags product. Agritech. 2013;32(4):362–369.
4. Huang Y-W, Liu Y, Dushenkov S, Ho C-T, Huang M-T. Anti-obesity effects of epigallocatechin-3-gallate, orange peel extract, black tea extract, caffeine and their combinations in a mouse model. J. Funct. Foods. 2009;1(3):304–310.
5. Geetha T, Garg A, Chopra K, Pal Kaur I. Delineation of antimutagenic activity of catechin, epicatechin and green tea extract. Mutat. Res. Mol. Mech. Mutagen. 2004;556(1–2):65–74.
6. Miura Y, Chiba T, Tomita I, Koizumi H, Miura S, Umegaki K, Hara Y, Ikeda M, Tomita T. Tea Catechins Prevent the Development of Atherosclerosis in Apoprotein E–Deficient Mice. J. Nutr. 2001;131(1):27–32.
7. Pramila K, Julius A. In Vitro Antioxidant effect of Green Tea Polyphenol Epigallocatechin-3-Gallate (EGCG) in Protecting Cardiovascular Diseases. Res. J. Pharm. Technol. 2019; 12(3): 1265.
8. Aladag H, Ercisli S, Yesil DZ, Gormez A, Yesil M. Antifungal activity of green tea leaves (Camellia sinensis L.) sampled in different harvest time. Pharmacogn. Mag. 2005;5(20):437.
9. Mochizuki M, Hasegawa N. (-)-epigallocatechin-3-gallate reduces experimental colon injury in rats by regulating macrophage and mast cell. Phyther. Res. 2010;24(S1):S120–S122.
10. Avadhani KS, Amirthalingam M, Reddy MS, Udupa N, Mutalik S. Development and Validation of RP-HPLC Method for Estimation of Epigallocatechin -3-gallate (EGCG) in Lipid based Nanoformulations. Res. J. Pharm. Technol. 2016;9(6):725.
11. Pastore MN, Kalia YN, Horstmann M, Roberts MS. Transdermal patches : history, development and pharmacology. Br. J. Pharmacol. 2015;14:2179–2209.
12. Saklar S, Ertas E, Ozdemir IS, Karadeniz B. Effects of different brewing conditions on catechin content and sensory acceptance in Turkish green tea infusions. J. Food Sci. Technol. 2015;52(10):6639–46.
13. Ashihara H, Crozier A. Caffeine: a well known but little mentioned compound in plant science. Trends Plant Sci. 2001;6(9):407–13.
14. El-Shahawi MS, Hamza A, Bahaffi SO, Al-Sibaai AA, Abduljabbar TN. Analysis of some selected catechins and caffeine in green tea by high performance liquid chromatography. Food Chem. 2012;134(4):2268–2275.
15. Kotani A, Takahashi K, Hakamata H, Kojima S, Kusu F. Attomole catechins determination by capillary liquid chromatography with electrochemical detection. Anal. Sci. 2007;23(2):157–163.
16. Vovk I, Simonovska B, Vuorela H. Separation of eight selected flavan-3-ols on cellulose thin-layer chromatographic plates. J. Chromatogr. A. 2005;1077(2):188–94.
17. Chen Q, Zhao J, Zhang H, Wang X. Feasibility study on qualitative and quantitative analysis in tea by near-infrared spectroscopy with multivariate calibration. Anal. Chim. Acta. 2006;572(1):77–84.
18. Chen Q, Zhao J, Lin H. Study on discrimination of Roast green tea (Camellia sinensis L.) according to geographical origin by FT-NIR spectroscopy and supervised pattern recognition. Spectrochim. Acta Part A Mol. Biomol. Spectrosc. 2009; 72(4): 845–850.
19. Baldermann S, Fleischmann P, Bolten M, Watanabe N, Winterhalter P, Ito Y. Centrifugal precipitation chromatography, a powerful technique for the isolation of active enzymes from tea leaves (Camellia sinensis). J. Chromatogr. A. 2009; 1216(19) :4263–7.
20. Koutelidakis AE, Argiri K, Serafini M, Proestos C, Komaitis M, Pecorari M, Kapsokefalou M, Kapsokefalou M. Green tea, white tea, and Pelargonium purpureum increase the antioxidant capacity of plasma and some organs in mice. Nutrition. 2009;25(4):453–458.
21. Yanagida A, Shoji A, Shibusawa Y, Shindo H, Tagashira M, Ikeda M, Ito Y. Analytical separation of tea catechins and food-related polyphenols by high-speed counter-current chromatography. J. Chromatogr. A. 2006;1112(1–2):195–201.
22. Schurek J, Portolés T, Hajslova J, Riddellova K, Hernández F. Application of head-space solid-phase microextraction coupled to comprehensive two-dimensional gas chromatography–time-of-flight mass spectrometry for the determination of multiple pesticide residues in tea samples. Anal. Chim. Acta. 2008;611(2):163–172.
23. Saito ST, Welzel A, Suyenaga ES, Bueno F. A method for fast determination of epigallocatechin gallate (EGCG), epicatechin (EC), catechin (C) and caffeine (CAF) in green tea using HPLC. Ciência e Tecnol. Aliment. 2006;26(2):394–400.
24. Proyong P, Weerapreeyakul N, Sripanidkulchai B. Validation of isocratic eluting and stepwise flow rate gradient for HPLC determination of cateohins, gallic acid and caffeine in tea. ScienceAsia. 2007;33(1):113–117.
25. Siva Kumar R, Kumar Nallasivan P, Saravanakumar S, Kandasamy C, Venkatnarayanan R. Simultaneous RP-HPLC estimation of nitazoxanide and ofloxacin in tablet dosage forms. Asian J. Res. Chem. 2009;2(41):43–45.
26. Battu PR, Reddy M. RP-HPLC method for simultaneous stimation of paracetamol and ibuprofen in tablets. Asian J. Res. Chem. 2009;2(1):70–72.
27. Manish BN, Jawaharlal DS, Nivrutti MH. Validated RP-HPLC method for simultaneous determination of amlodipine besylate and hydrochlorothiazide from pharmaceutical preparation and biological samples. Res. J. Pharm. Technol. 2009;2(3):482–486.
28. Shradanjali Singh M, Kirti Topagi S, Mrinalini Damle S. A validated RP-HPLC for simultaneous determination of ramipril and valsartan in pharmaceutical dosage form. Res. J. Pharm. Technol. 2009;2(4):749–752.
29. Verma J, Jain D, Jain N, Sharad Padey P, Jain DK. Simultaneous estimation of lamivudine, stavudine, and nevirapine by RP-HPLC in tablet formulation. Res. J. Pharm. Technol. 2010;3(2):490–493.
30. Indrati O, Martien R, Rohman A, Nugroho AK. Emplyment of ATR-FTIR and HPLC-UV method for detction and quantification of andrographolide. Int. J. Appl. Pharm. 2018;10(6):135.
31. Setyawan EI, Rohman A, Setyowati EP, Nugroho AK. Application of factorial design on the extraction of green tea leaves (Camellia sinensis L.). J. Appl. Pharm. Sci. 2018;8(4):131–138.
32. Setyawan EI, Setyowati EP, Rohman A, Nugroho AK. Central composite design for optimizing extraction of EGCG from greean tea leaf (Camellia sinensis L.). Int. J. Appl. Pharm. 2018;10(6):211–216.
33. Singh M, Kashkhedikar SG, Soni L, Garg A, Gandhi T, Patel A. Development of RP-HPLC method for estimation of carvedilol in tablet formulations. Res. J. Pharm. Technol. 2008;1(1):18–21.
34. Rajavel R, Ganesh M, Jagadeeswaran M, Srinivasan K, Valarmathi J, Sivakumar T. RP-HPLC method for the simultaneous determination of aspirin , atorvastatin and pioglitazone in capsule dosage form. Asian J. Res. Chem. 2008;1(1):40–42.
35. Jain N, Jain R, Swami H, Jain DK. RP-HPLC method for simultaneous estimation of simvastatin and ezetimibe in bulk drug and its combined dosage form. Asian J. Res. Chem. 2008;1(1):29–31.
36. Krupkova O, Ferguson SJ, Wuertz-Kozak K. Stability of (−)-epigallocatechin gallate and its activity in liquid formulations and.pdf. J. Nutr. Biochem. 2016;37:1–12.
37. Wang H, Helliwell K. Epimerisation of catechins in green tea infusions. Food Chem. 2000;70(3):337–344.