Copyright (c) 2024 Ms Dwaipee De, SONALI RAY, Dr Mayukh Deb
This work is licensed under a Creative Commons Attribution 4.0 International License.
Comprehensive Isolation and Characterization of Theaflavins, Thearubigins and Caffeine as Bioactive Compounds from Darjeeling Black Tea
Corresponding Author(s) : Sonali Ray
Asian Journal of Chemistry,
Vol. 36 No. 10 (2024): Vol 36 Issue 10, 2024
Abstract
The present study aimed to isolate and characterize three major bioactive components viz. theaflavins, thearubigins and caffeine from Darjeeling black tea, which contribute significantly to the cup characteristics and health benefits of black tea. Theaflavins were isolated using Soxhlet extraction followed by column chromatography, HPLC and UV spectroscopy. HPLC showed effective separation of four major theaflavins. Thearubigins were isolated using liquid-liquid extraction and characterized through NMR, FTIR-ATR and UV spectroscopic techniques. The NMR spectral studies suggested thearubigins to be polymeric oxidation products of catechin groups, while FTIR spectral studies indicated similarity with proanthocyanidin-like polymers. Caffeine was isolated via a solid-liquid extraction followed by liquid-liquid extraction and identified using TLC, NMR, UV and SEM techniques. TLC showed an Rf value of 0.65, which is consistent with standard caffeine. The NMR confirmed the purity of isolated caffeine and SEM revealed its needle-shaped crystal morphology. The results show that these bioactive compounds may be efficiently isolated and characterized using chromatographic and spectroscopic methods.
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- K.S. Khan, BMJ, 358, j3119 (2017); https://doi.org/10.1136/bmj.j3119
- B. Alipoor and A.H. Rad, Asian J. Clin. Nutr., 4, 1 (2011); https://doi.org/10.3923/ajcn.2012.1.15
- A. Imran, M.U. Arshad, S. Mehmood, R.S. Ahmed, M.S. Butt, A. Ahmed, M. Imran, M.S. Arshad, N. Faiza, I. Haq, S.A.S. Chatha, M. Shahbaz, Q. Ali and M. Waseem, Polyphenols, 18, 25 (2018); https://doi.org/10.5772/intechopen.75933
- G.H. Yassin, J.H. Koek and N. Kuhnert, Food Chem., 180, 272 (2015); https://doi.org/10.1016/j.foodchem.2015.01.108
- M.E. Harbowy, D.A. Balentine, A.P. Davies and Y. Cai, Crit. Rev. Plant Sci., 16, 415 (1997); https://doi.org/10.1080/07352689709701956
- C.S. Yang, G. Chen and Q. Wu, J. Tradit. Complement. Med., 4, 17 (2014); https://doi.org/10.4103/2225-4110.124326
- M. Da Silva Pinto, Food Res. Int., 53, 558 (2013); https://doi.org/10.1016/j.foodres.2013.01.038
- Y.C. Yang, F.H. Lu, J.S. Wu, C.H. Wu and C.J. Chang, Arch. Intern. Med., 164, 1534 (2004); https://doi.org/10.1001/archinte.164.14.1534
- J.A. Vinson, Biofactors, 13, 127 (2000); https://doi.org/10.1002/biof.5520130121
- S.A. Khan, S.S. Chatterjee and V. Kumar, Life Sci., 148, 53 (2016); https://doi.org/10.1016/j.lfs.2016.02.032
- K. Boehm, F. Borrelli, E. Ernst, G. Habacher, S.K. Hung, S. Milazzo and M. Horneber, Cochrane Database Syst. Rev., 3, CD005004 (2009); https://doi.org/10.1002/14651858.CD005004.pub2
- J. Minatti, E. Wazlawik, M.A. Hort, F.L. Zaleski, R.M. Ribeiro-do-Valle, M. Maraschin and E.L. da Silva, Nutr. Res., 32, 684 (2012); https://doi.org/10.1016/j.nutres.2012.08.003
- S. Ding, J. Jiang, P. Yu, G. Zhang, G. Zhang and X. Liu, PLoS One, 12, e0181666 (2017); https://doi.org/10.1371/journal.pone.0181666
- C.A. Larsen, R.H. Dashwood and W.H. Bisson, Pharmacol. Res., 62, 457 (2010); https://doi.org/10.1016/j.phrs.2010.07.010
- Y. Sheng, Y. Sun, Y. Tang, Y. Yu, J. Wang, F. Zheng, Y. Li and Y. Sun, Front. Pharmacol., 14, 1144878 (2023); https://doi.org/10.3389/fphar.2023.1144878
- J. Hong, T.J. Smith, C.T. Ho, D.A. August and C.S. Yang, Biochem. Pharmacol., 62, 1175 (2001); https://doi.org/10.1016/S0006-2952(01)00767-5
- M.C. Menet, S. Sang, C.S. Yang, C.T. Ho and R.T. Rosen, J. Agric. Food Chem., 52, 2455 (2004); https://doi.org/10.1021/jf035427e
- D. De and S. Ray, Int. J. Pharm. Sci. Res., 13, 2320 (2022); https://doi.org/10.13040/IJPSR.0975-8232.13(4).1508-23
- M. Takemoto and H. Takemoto, Molecules, 23, 918 (2018); https://doi.org/10.3390/molecules23040918
- E.A.H. Roberts and R.F. Smith, Analyst, 86, 94 (1961); https://doi.org/10.1039/AN9618600094
- J.W. Drynan, M.N. Clifford, J. Obuchowicz and N. Kuhnert, J. Agric. Food Chem., 60, 4514 (2012); https://doi.org/10.1021/jf205125y
- N. Kuhnert, Arch. Biochem. Biophys., 501, 37 (2010); https://doi.org/10.1016/j.abb.2010.04.013
- T. Ozawa, M. Kataoka, K. Morikawa and O. Negishi, Biosci. Biotechnol. Biochem., 60, 2023 (1996); https://doi.org/10.1271/bbb.60.2023
- G.H. Yassin, J.H. Koek, S. Jayaraman and N. Kuhnert, J. Agric. Food Chem., 62, 9848 (2014); https://doi.org/10.1021/jf502220c
- J.E. Berkowitz, P. Coggon and G.W. Sanderson, Phytochemistry, 10, 2271 (1971); https://doi.org/10.1016/S0031-9422(00)89866-0
- E.A.H. Roberts, J. Sci. Food Agric., 9, 212 (1958); https://doi.org/10.1002/jsfa.2740090405
- R.G. Bailey, H.E. Nursten and I. McDowell, J. Chromatogr. A, 542, 115 (1991); https://doi.org/10.1016/S0021-9673(01)88752-5
- A.G. Brown, W.B. Eyton, A. Holmes and W.D. Ollis, Nature, 221, 742 (1969a); https://doi.org/10.1038/221742a0
- A.G. Brown, W.B. Eyton, A. Holmes and W.D. Ollis, Phytochemistry, 8, 2333 (1969b); https://doi.org/10.1016/S0031-9422(00)88151-0
- A. Mumin, K.F. Akhter, Z. Abedin and Z. Hossain, Malaysian J. Chem., 8, 45 (2006).
- S.M. Tarka and W.J. Hurst, Progr. Clin. Biol. Res., 158, 9 (1984).
- K. Zulak, D. Liscombe, H. Ashihara and P. Facchini, Alkaloids: Plant Secondary Metabolism in Diet and Human Health, Oxford: Blackwell Publishing, pp. 102-36 (2006).
- J. de Paula Lima and A. Farah, Caffeine and Minor Methylxanthines in Coffee, In: Coffee: Production, Quality and Chemistry, The Royal Society of Chemistry, Chap. 23, pp. 543-564 (2019).
- F.A. Al-Bayati and M.J. Mohammed, Int. J. Green Pharm., 3, 52 (2009); https://doi.org/10.4103/0973-8258.49375
- G.V. Siva, G. Kirubananthan and C. Alagarasan, Int. J. Curr. Biotech., 5, 1 (2017).
- S. Ray, T. Samanta, A. Mitra and B. De, Curr. Nutr. Food Sci., 10, 181 (2014); https://doi.org/10.2174/1573401310666140529205646
- G. Navarra, M. Moschetti, V. Guarrasi, M.R. Mangione, V. Militello and M. Leone, J. Chem., 2017, 6435086 (2017); https://doi.org/10.1155/2017/6435086
- A. Misra, R. Chattopadhyay, S. Banerjee, D.J. Chattopadhyay and I.B. Chatterjee, J. Nutr., 133, 2622 (2003); https://doi.org/10.1093/jn/133.8.2622
- A. Chaugule, H. Patil, S. Pagariya and P. Ingle, J. Int. Adv. Res. Chem. Sci., 6, 11 (2019); https://doi.org/10.20431/2349-0403.0609002
- T. Baishya, P. Das, G.J. Ashraf, T.K. Dua, P. Paul, G. Nandi, M. Bhattacharya and R. Sahu, Z. Naturforsch. C J. Biosci., 78, 235 (2023); https://doi.org/10.1515/znc-2022-0174
- L. Horner, S. Göwecke and W. Dürckheimer, Chem. Ber., 94, 1276 (1961); https://doi.org/10.1002/cber.19610940517
- L. Horner, W. Dürckheimer, K.H. Weber and K. Dölling, Chem. Ber., 97, 312 (1964); https://doi.org/10.1002/cber.19640970203
- T. Tanaka, K. Inoue, Y. Betsumiya, C. Mine and I. Kouno, J. Agric. Food Chem., 49, 5785 (2001); https://doi.org/10.1021/jf010842x
- K. Zhu, J. Ouyang, J. Huang and Z. Liu, Crit. Rev. Food Sci. Nutr., 61, 1556 (2021); https://doi.org/10.1080/10408398.2020.1762161
- R.G. Bailey, H.E. Nursten and I. McDowell, J. Sci. Food Agric., 59, 365 (1992); https://doi.org/10.1002/jsfa.2740590314
- L.Y. Foo, Phytochemistry, 20, 1397 (1981); https://doi.org/10.1016/0031-9422(81)80047-7
- T. Tanaka, M. Yasumatsu, M. Hirotani, Y. Matsuo, N. Li, H.T. Zhu, Y. Saito, K. Ishimaru and Y.J. Zhang, Food Chem., 370, 131326 (2022); https://doi.org/10.1016/j.foodchem.2021.131326
- J. Sitkowski, L. Stefaniak, L. Nicol, M.L. Martin, G.J. Martin and G.A. Webb, Spectrochim. Acta A Mol. Biomol. Spectrosc., 51, 839 (1995); https://doi.org/10.1016/0584-8539(94)00192-E
- D. Habtamu and A. Belay, Food Sci. Nutr., 8, 4757 (2020); https://doi.org/10.1002/fsn3.1723
- A. Sarfraz, A. Simo, R. Fenger, W. Christen, K. Rademann, U. Panne and F. Emmerling, Cryst. Growth Des., 12, 583 (2012); https://doi.org/10.1021/cg101358q
References
K.S. Khan, BMJ, 358, j3119 (2017); https://doi.org/10.1136/bmj.j3119
B. Alipoor and A.H. Rad, Asian J. Clin. Nutr., 4, 1 (2011); https://doi.org/10.3923/ajcn.2012.1.15
A. Imran, M.U. Arshad, S. Mehmood, R.S. Ahmed, M.S. Butt, A. Ahmed, M. Imran, M.S. Arshad, N. Faiza, I. Haq, S.A.S. Chatha, M. Shahbaz, Q. Ali and M. Waseem, Polyphenols, 18, 25 (2018); https://doi.org/10.5772/intechopen.75933
G.H. Yassin, J.H. Koek and N. Kuhnert, Food Chem., 180, 272 (2015); https://doi.org/10.1016/j.foodchem.2015.01.108
M.E. Harbowy, D.A. Balentine, A.P. Davies and Y. Cai, Crit. Rev. Plant Sci., 16, 415 (1997); https://doi.org/10.1080/07352689709701956
C.S. Yang, G. Chen and Q. Wu, J. Tradit. Complement. Med., 4, 17 (2014); https://doi.org/10.4103/2225-4110.124326
M. Da Silva Pinto, Food Res. Int., 53, 558 (2013); https://doi.org/10.1016/j.foodres.2013.01.038
Y.C. Yang, F.H. Lu, J.S. Wu, C.H. Wu and C.J. Chang, Arch. Intern. Med., 164, 1534 (2004); https://doi.org/10.1001/archinte.164.14.1534
J.A. Vinson, Biofactors, 13, 127 (2000); https://doi.org/10.1002/biof.5520130121
S.A. Khan, S.S. Chatterjee and V. Kumar, Life Sci., 148, 53 (2016); https://doi.org/10.1016/j.lfs.2016.02.032
K. Boehm, F. Borrelli, E. Ernst, G. Habacher, S.K. Hung, S. Milazzo and M. Horneber, Cochrane Database Syst. Rev., 3, CD005004 (2009); https://doi.org/10.1002/14651858.CD005004.pub2
J. Minatti, E. Wazlawik, M.A. Hort, F.L. Zaleski, R.M. Ribeiro-do-Valle, M. Maraschin and E.L. da Silva, Nutr. Res., 32, 684 (2012); https://doi.org/10.1016/j.nutres.2012.08.003
S. Ding, J. Jiang, P. Yu, G. Zhang, G. Zhang and X. Liu, PLoS One, 12, e0181666 (2017); https://doi.org/10.1371/journal.pone.0181666
C.A. Larsen, R.H. Dashwood and W.H. Bisson, Pharmacol. Res., 62, 457 (2010); https://doi.org/10.1016/j.phrs.2010.07.010
Y. Sheng, Y. Sun, Y. Tang, Y. Yu, J. Wang, F. Zheng, Y. Li and Y. Sun, Front. Pharmacol., 14, 1144878 (2023); https://doi.org/10.3389/fphar.2023.1144878
J. Hong, T.J. Smith, C.T. Ho, D.A. August and C.S. Yang, Biochem. Pharmacol., 62, 1175 (2001); https://doi.org/10.1016/S0006-2952(01)00767-5
M.C. Menet, S. Sang, C.S. Yang, C.T. Ho and R.T. Rosen, J. Agric. Food Chem., 52, 2455 (2004); https://doi.org/10.1021/jf035427e
D. De and S. Ray, Int. J. Pharm. Sci. Res., 13, 2320 (2022); https://doi.org/10.13040/IJPSR.0975-8232.13(4).1508-23
M. Takemoto and H. Takemoto, Molecules, 23, 918 (2018); https://doi.org/10.3390/molecules23040918
E.A.H. Roberts and R.F. Smith, Analyst, 86, 94 (1961); https://doi.org/10.1039/AN9618600094
J.W. Drynan, M.N. Clifford, J. Obuchowicz and N. Kuhnert, J. Agric. Food Chem., 60, 4514 (2012); https://doi.org/10.1021/jf205125y
N. Kuhnert, Arch. Biochem. Biophys., 501, 37 (2010); https://doi.org/10.1016/j.abb.2010.04.013
T. Ozawa, M. Kataoka, K. Morikawa and O. Negishi, Biosci. Biotechnol. Biochem., 60, 2023 (1996); https://doi.org/10.1271/bbb.60.2023
G.H. Yassin, J.H. Koek, S. Jayaraman and N. Kuhnert, J. Agric. Food Chem., 62, 9848 (2014); https://doi.org/10.1021/jf502220c
J.E. Berkowitz, P. Coggon and G.W. Sanderson, Phytochemistry, 10, 2271 (1971); https://doi.org/10.1016/S0031-9422(00)89866-0
E.A.H. Roberts, J. Sci. Food Agric., 9, 212 (1958); https://doi.org/10.1002/jsfa.2740090405
R.G. Bailey, H.E. Nursten and I. McDowell, J. Chromatogr. A, 542, 115 (1991); https://doi.org/10.1016/S0021-9673(01)88752-5
A.G. Brown, W.B. Eyton, A. Holmes and W.D. Ollis, Nature, 221, 742 (1969a); https://doi.org/10.1038/221742a0
A.G. Brown, W.B. Eyton, A. Holmes and W.D. Ollis, Phytochemistry, 8, 2333 (1969b); https://doi.org/10.1016/S0031-9422(00)88151-0
A. Mumin, K.F. Akhter, Z. Abedin and Z. Hossain, Malaysian J. Chem., 8, 45 (2006).
S.M. Tarka and W.J. Hurst, Progr. Clin. Biol. Res., 158, 9 (1984).
K. Zulak, D. Liscombe, H. Ashihara and P. Facchini, Alkaloids: Plant Secondary Metabolism in Diet and Human Health, Oxford: Blackwell Publishing, pp. 102-36 (2006).
J. de Paula Lima and A. Farah, Caffeine and Minor Methylxanthines in Coffee, In: Coffee: Production, Quality and Chemistry, The Royal Society of Chemistry, Chap. 23, pp. 543-564 (2019).
F.A. Al-Bayati and M.J. Mohammed, Int. J. Green Pharm., 3, 52 (2009); https://doi.org/10.4103/0973-8258.49375
G.V. Siva, G. Kirubananthan and C. Alagarasan, Int. J. Curr. Biotech., 5, 1 (2017).
S. Ray, T. Samanta, A. Mitra and B. De, Curr. Nutr. Food Sci., 10, 181 (2014); https://doi.org/10.2174/1573401310666140529205646
G. Navarra, M. Moschetti, V. Guarrasi, M.R. Mangione, V. Militello and M. Leone, J. Chem., 2017, 6435086 (2017); https://doi.org/10.1155/2017/6435086
A. Misra, R. Chattopadhyay, S. Banerjee, D.J. Chattopadhyay and I.B. Chatterjee, J. Nutr., 133, 2622 (2003); https://doi.org/10.1093/jn/133.8.2622
A. Chaugule, H. Patil, S. Pagariya and P. Ingle, J. Int. Adv. Res. Chem. Sci., 6, 11 (2019); https://doi.org/10.20431/2349-0403.0609002
T. Baishya, P. Das, G.J. Ashraf, T.K. Dua, P. Paul, G. Nandi, M. Bhattacharya and R. Sahu, Z. Naturforsch. C J. Biosci., 78, 235 (2023); https://doi.org/10.1515/znc-2022-0174
L. Horner, S. Göwecke and W. Dürckheimer, Chem. Ber., 94, 1276 (1961); https://doi.org/10.1002/cber.19610940517
L. Horner, W. Dürckheimer, K.H. Weber and K. Dölling, Chem. Ber., 97, 312 (1964); https://doi.org/10.1002/cber.19640970203
T. Tanaka, K. Inoue, Y. Betsumiya, C. Mine and I. Kouno, J. Agric. Food Chem., 49, 5785 (2001); https://doi.org/10.1021/jf010842x
K. Zhu, J. Ouyang, J. Huang and Z. Liu, Crit. Rev. Food Sci. Nutr., 61, 1556 (2021); https://doi.org/10.1080/10408398.2020.1762161
R.G. Bailey, H.E. Nursten and I. McDowell, J. Sci. Food Agric., 59, 365 (1992); https://doi.org/10.1002/jsfa.2740590314
L.Y. Foo, Phytochemistry, 20, 1397 (1981); https://doi.org/10.1016/0031-9422(81)80047-7
T. Tanaka, M. Yasumatsu, M. Hirotani, Y. Matsuo, N. Li, H.T. Zhu, Y. Saito, K. Ishimaru and Y.J. Zhang, Food Chem., 370, 131326 (2022); https://doi.org/10.1016/j.foodchem.2021.131326
J. Sitkowski, L. Stefaniak, L. Nicol, M.L. Martin, G.J. Martin and G.A. Webb, Spectrochim. Acta A Mol. Biomol. Spectrosc., 51, 839 (1995); https://doi.org/10.1016/0584-8539(94)00192-E
D. Habtamu and A. Belay, Food Sci. Nutr., 8, 4757 (2020); https://doi.org/10.1002/fsn3.1723
A. Sarfraz, A. Simo, R. Fenger, W. Christen, K. Rademann, U. Panne and F. Emmerling, Cryst. Growth Des., 12, 583 (2012); https://doi.org/10.1021/cg101358q