Korean Journal of Food Science and Technology (한국식품과학회지)

Korean Society of Food Science and Technology (한국식품과학회)
권호 표지

Analytical Method for Methylxanthin, Catechin, and Theaflavin Determinations in Korean Commercial Teas by HPLC

차에 함유된 methylxanthin류, catechin류 및 theaflavin류의 HPLC에 의한 동시분석법

  • Published : 2006.02.01
Download PDF
⟨ Previous Next ⟩

Abstract

Method for separation and quantification of methylxanthins, catechins, and theaflavins in Korean commercial teas (green, oolong, and black teas) was developed using reversed phase high-performance liquid chromatography (HPLC). After extraction with hot water, tea compounds were separated on Inertsil ODS-3v $(5\;{\mu}m)$ column, eluted with gradient of 7% acetonitrile and 93% of 20 mM phosphate buffer mixture for 7 min. Column effluent was monitored at 270 nm. This technique was effective for analyses of m methylxanthins, catechins, and theaflavins in teas and biological samples. In green and oolong teas, two kinds of methylxanthins and 7 of catechins were identified, whereas 4 theaflavins were only identified in black tea. Among seven catechins in green and oolong teas, EGCG showed highest amount, whereas ECG was highest in black tea. (theaflavins were found only in black teas) In all teas, theobromine content was lower than that of caffeine.

본 연구는 차에 함유되어 있는 특유의 정미성분으로 methylxantin류, catechin류 및 theaflavin류의 총 13종의 화합물을 HPLC를 이용한 동시분석법을 확립함과 동시에 이 분석법을 이용하여 한국에서 사판되고 있는 녹차, 홍차 및 우롱차에 함유되어 있는 주요 성분을 분석하였으며 그 결과는 다음과 같다. HPLC분석조건은 역상(ODS)column을 이용하여 acetonitrile: 20 mM 인산완충액의 solvent 중 acetonitrile의 농도를 처음 단계 7%에서 최종 40%까지 단계적으로 변화시켜 분당 1mL씩 용출시켜 분석하였다. Column 온도는 효과적으로 각 성분을 분리시키기 위하여 정확하게 $30^{\circ}C$로 설정하였으며 파장은 270 nm에서 분석하였다. 또한 차에 함유되어 있는 methylxantin류, catechin류 및 theaflavin류의 총 13종의 화합물을 분석한 결과, 한 시료당 90분이 소요되었으며 재현성과 정량성이 뛰어나 13종의 화합물이 완전하게 분리되었다. 시판 녹차, 홍차, 우롱차를 분석한 결과, 녹차와 우롱차는 2종류의 methylxantin류와 7종류의 catechin류는 검출되었으나 홍차에서 검출된 4종류의 theaflavin류는 전혀 나타나지 않았다. 이러한 결과는 차의 품질 관리 및 소비자의 차 선택에 있어 중요한 기초자료가 될 것으로 사료된다.

Keywords

References

  1. Caudle AG, Gu Y, Bell LN. Caffeine and theobromine contents of ready-to-eat chocolate cereals. J. Am. Diet. Assoc. 100: 690-692 (2000) https://doi.org/10.1016/S0002-8223(00)00200-5
  2. Martinez LL, Alba PLL, Campos RG, Rodriguez LMD. Simultaneous determination of methylxanthines in coffees and teas by UV-Vis spectrometry and partial least squares. Anal. Chim. Acta. 493: 83-94 (2003) https://doi.org/10.1016/S0003-2670(03)00862-6
  3. Blauch JL, Tarka SM, HPLC determination of caffeine and theobromine in coffee, tea, and instant hot cocoa mixes. J. Food Sci. 48: 745-750 (1983) https://doi.org/10.1111/j.1365-2621.1983.tb14888.x
  4. Craig WJ, Nguyen TT. Caffeine and theobromine levels in cocoa and carob products. J. Food Sci. 49: 302-305 (1984) https://doi.org/10.1111/j.1365-2621.1984.tb13737.x
  5. Wang LF, Kim DM, Lee CY. Interaction of flavanols in green tea extract during heat processing and storage. Food Sci. Biotechnol. 11: 608-612 (2002)
  6. Obanda PM, Owuor P, Mang'oka R, Kavoi MM. Changes in thearubigin fractions and theaflavin levels due to variations in processing conditions and their influence on black tea liquor brightness and total colour. Food Chem. 85: 163-173 (2004) https://doi.org/10.1016/S0308-8146(02)00183-8
  7. Obanda PM, Owuor P, Mang'oka R. Changes in the chemical and sensory quality parameters of black tea due to variations of fermentation time and temperature. Food Chem. 75: 395-404 (2001) https://doi.org/10.1016/S0308-8146(01)00223-0
  8. Luczaj M, Skrzydlewska E. Antioxidant properties of black tea in alcohol intoxication. Food Chem. Toxicol. 42: 2045-2051 (2004) https://doi.org/10.1016/j.fct.2004.08.009
  9. Majchrzak D, Mitter S, Elmadfa I. The effect of ascorbic acid on total antioxidant activity of black and green teas. Food Chem. 88: 447-451 (2004) https://doi.org/10.1016/j.foodchem.2004.01.058
  10. Yam TS, Shah S, Hamilton-Miller JMT. Microbiological activity of whole and fractionated crude extracts of tea (Camellia sinensis), and of tea components. FEMS Microbiol. Lett. 152: 169-174 (1997) https://doi.org/10.1111/j.1574-6968.1997.tb10424.x
  11. Yoda Y, Hu ZQ, Zho WH, Shimamura T. Different susceptibilities of Staphylococcus and Gram-negative rods to epigallocatichin gallate. J. Infect. Chemother. 10: 55-58 (2004) https://doi.org/10.1007/s10156-003-0284-0
  12. Urabe K, Nadamoto T, Kawamura M, Yasumoto K. Effects of Houttuyniae cordata and refinery final molasses on the development of offensive odor in porcine small intestine during storage. J. Nutr. Sci. Vitaminol (Tokyo). 40: 63-71 (1994) https://doi.org/10.3177/jnsv.40.63
  13. Wang LF, Zhang HY. A theoretical study of the different radical-scavenging activities of catechin, quercetin and a rationally designed planar catechin. Bioorg. Chem. 33: 108-115 (2005) https://doi.org/10.1016/j.bioorg.2005.01.002
  14. Jin G, Yoshioka H. Synthesis of lipophilic poly-Iauroyl-(+)-catechins and radical-scavenging activity. Biosci. Biotechnol. Biochem. 69: 440-447 (2005) https://doi.org/10.1271/bbb.69.440
  15. Ikeda I, Kobayashi M, Hamada T, Tsuda K, Goto H, Imaizumi K, Nozawa A, Sugimoto A, Kakuda T. Heat-epimerized tea catechins rich in gallocatechin gallate and catechin gallate are more effective to inhibit cholesterol absorption than tea catechins rich in epigallocatechin gallate and epicatechin gallate. J. Agric. Food Chem. 51: 7303-7307 (2003) https://doi.org/10.1021/jf034728l
  16. Raederstorff DG, Schlachter MF, Elste V, Weber P. Effect of EGCG on lipid absorption and plasma lipid levels in rats. J. Nutr. Biochem. 14: 326-32 (2003) https://doi.org/10.1016/S0955-2863(03)00054-8
  17. Rizvi SI, Zaid MA, Anis R, Mishra N. Protective role of tea catechins against oxidation-induced damage of type 2 diabetic erythrocytes. Clin. Exp. Pharmacol. Physiol. 2: 70-75 (2005)
  18. Crespy V, Williamson G. A review of the health effects of green tea catechins in in vivo animal models. J. Nutr. Rev. 134: 3431-3440 (2004) https://doi.org/10.1093/jn/134.12.3431S
  19. Villar A, Terencio MC, Paya M. Hypotensive effect of Rhamnus lycioides extracts. Ethnopharmacol. 16: 269-273 (1986) https://doi.org/10.1016/0378-8741(86)90093-0
  20. Negishi H, Xu JW, Ikeda K, Njelekela M, Nara Y, Yamori Y. Black and green tea polyphenols attenuate blood pressure increases in stroke-prone spontaneously hypertensive rats. J. Nutr. 134: 38-42 (2004)
  21. Ramos S, Alia M, Bravo L, Goya L. Comparative effects of food-derived polyphenols on the viability and apoptosis of a human hepatoma cell line (HepG2). J. Agric. Food Chem. 53: 1271-1280 (2005) https://doi.org/10.1021/jf0490798
  22. Henning SM, Niu Y, Lee NH, Thames GD, Minutti RR, Wang H, Go VL, Heber D. Bioavailability and antioxidant activity of tea flavanols after consumption of green tea, black tea, or a green tea extract supplement. Am. J. Clin. Nutr. 80: 1558-1564 (2004) https://doi.org/10.1093/ajcn/80.6.1558
  23. Nikaidou S, Ishizuka M, Maeda Y, Hara Y, Kazusaka A, Fujita S. Effect of catechins on mutagenesis of Salmonella typhimurium TA 102 elicited by tert-butyl hydroperoxide (t-BuOOH). J. Vet. Med. Sci. 67: 137-138 (2005) https://doi.org/10.1292/jvms.67.137
  24. Geetha T, Garg A, Chopra K, Kaur I. Delineation of antimutagenic activity of catechin, epicatechin and green tea extract. Mutat. Res. 22: 65-74 (2004)
  25. Sachinidis A, Seul C, Seedwald S, Ahn H, Ko Y, Vetter H. Green tea compounds inhibit tyrosine phosphorylation of PDGF beta-receptor and transformation of A 172 human glioblastoma. FEBS. Lett. 471: 51-55 (2000) https://doi.org/10.1016/S0014-5793(00)01360-0
  26. Nishitani E, Sagesaka YM. Simultaneous determination of catechins, caffeine and other phenolic compounds in tea using new HPLC method. J. Food Comp. Analysis 17: 675-685 (2004) https://doi.org/10.1016/j.jfca.2003.09.009
  27. Aucamp JP, Hara Y, Apostolides Z. Simultaneous analysis of tea catechins, caffeine, gallic acid, theanine and ascorbic acid by micellar electrokinetic capillary chromatography. J. Chromatogra. A. 876: 235-242 (2000) https://doi.org/10.1016/S0021-9673(00)00145-X
  28. Goto T, Yoshida Y, Kiso M, Nagashima K. Simultaneous analysis of individual catechins and caffeine in green tea. J. Chromatogra. A. 749: 295-299 (1996) https://doi.org/10.1016/0021-9673(96)00456-6
  29. Wang H, Provan GJ, Helliwell K. HPLC determination of catechins in tea leaves and tea extractions using relative response factors. Food Chem. 81: 307-312 (2003) https://doi.org/10.1016/S0308-8146(02)00510-1
  30. Lee YJ, Ahn MS, Oh WT. A Study on the catechins content and antioxidative effect of various solvent extracts of green, oolng and black tea. J. Food Hyg. Safety 13: 370-376 (1998)