Simultaneous Determination of Epi-Catechins and Their Oxidized Dimers, Gallic Acid and Caffeine in Tea by HPLC

ZHANG Kaixin; KONG Xianghui; XU Weiqi; CHEN Guijie; ZENG Xiaoxiong

doi:10.13386/j.issn1002-0306.2022120093

Volume 44 Issue 19

Oct. 2023

Turn off MathJax

Article Contents

Abstract

References

Science and Technology of Food Industry > 2023 > 44(19): 366-373. > DOI: 10.13386/j.issn1002-0306.2022120093

ZHANG Kaixin, KONG Xianghui, XU Weiqi, et al. Simultaneous Determination of Epi-Catechins and Their Oxidized Dimers, Gallic Acid and Caffeine in Tea by HPLC[J]. Science and Technology of Food Industry, 2023, 44(19): 366−373. (in Chinese with English abstract). doi: 10.13386/j.issn1002-0306.2022120093.

Citation:

PDF (1751 KB)

Simultaneous Determination of Epi-Catechins and Their Oxidized Dimers, Gallic Acid and Caffeine in Tea by HPLC

1.
College of Food Science and Technology, Nanjing Agricultural University, Nanjing 210095, China
2.
State Key Laboratory of Tea Plant Biology and Utilization, Anhui Agricultural University, Hefei 230036, China

More Information

Received Date: December 11, 2022
Available Online: July 26, 2023

Graphical Abstract

Abstract

Abstract

Objective: The present study was aimed to develop the simultaneous determination of four kinds of epi-catechins ((-)-epicatechin, (-)-epicatechin gallate, (-)-epigallocatechin and (-)-epigallocatechin gallate) and their oxidized dimers (four kinds of theaflavin (TF): theaflavin, theaflavin-3-gallate, theaflavin-3'-gallate and theaflavin-3,3'-digallate, three kinds of theasinensin (TS): TSA, TSB and TSC), gallic acid and caffeine by high performance liquid chromatography (HPLC). Methods: Firstly, TSA, TSB and TSC were chemically synthesized. Then, the gradient elution conditions for HPLC analysis were obtained according to the reported methods with some modifications. Chromatographic separation was achieved on a Cosmosil 5C₁₈-AR-Ⅱ column (250 mm×4.6 mm, 5 μm). Ultrapure water (eluent A), 50 mmol/L H₃PO₄ (eluent B) and acetonitrile (eluent C) were used as the mobile phases. The DAD acquisition wavelength was set at 280 nm. The flow rate was 0.8 mL/min and oven temperature was set at 30 ℃. The volume of injection was 20 μL. Finally, the conents of these componds mentioned above in green, oolong, black and dark teas were determined by the developed HPLC method. Results: Under the optimized conditions, 4 kinds of epicatechins, 4 kinds of theaflavins, 3 kinds of theasinensins, gallic acid and caffeine could be separated within 66 min. At the same time, in the range of 0.01~1.00 mg/mL, the substances and their peak areas showed a good linear relationship (R², 0.9904~0.9966). The relative standard of deviation (RSD) for repeatability was 1.034%~4.008%. The RSD of the precision and the stability were 1.882%~4.835% and 0.349%~3.994%, respectively. The recovery test of three concentrations was in the range of 90.106%~107.331% and the RSD was less than 3.268%. In adition, it was found that the contents of these 13 compounds were significantly different among different tea samples, and all the detected values were within the effective determination range of the standard curve. Conclusion: The developend method is simple, accurate, and reproducible, it can be used for the simultaneous determination of epi-catechins, theaflavins, theasinensins, gallic acid, and caffeine in tea samples.
- tea,
- HPLC,
- epicatechins,
- theaflavins,
- theasinensins,
- caffeine

FullText(HTML)

References (31)

References

[1]	桂安辉, 高士伟, 叶飞, 等. 不同产地扁形绿茶的品质成分差异分析[J]. 食品工业科技,2020,41(20):218−223. [GUI A H, GAO S W, YE F, et al. Differential analysis of quality components of flat green tea from different producing areas[J]. Science and Technology of Food Industry,2020,41(20):218−223. GUI A H, GAO S W, YE F, et al. Differential analysis of quality components of flat green tea from different producing areas[J]. Science and Technology of Food Industry, 2020, 41(20): 218-223.
[2]	XIAO Y, ZHONG K, BAI J R, et al. Insight into effects of isolated Eurotium cristatum from Pingwu Fuzhuan brick tea on the fermentation process and quality characteristics of Fuzhuan brick tea[J]. Journal of the Science of Food and Agriculture,2020,100(9):3598−3607. doi: 10.1002/jsfa.10353
[3]	宋楚君, 范方媛, 龚淑英, 等. 不同产地红茶的滋味特征及主要贡献物质[J]. 中国农业科学,2020,53(2):383−394. [SONG C J, FAN F Y, GONG S Y, et al. Taste characteristic and main contributing compounds of different origin black tea[J]. Scientia Agricultura Sinica,2020,53(2):383−394. doi: 10.3864/j.issn.0578-1752.2020.02.012 SONG C J, FAN F Y, GONG S Y, et al. Taste characteristic and main contributing compounds of different origin black tea[J]. Scientia Agricultura Sinica, 2020, 53(2): 383-394. doi: 10.3864/j.issn.0578-1752.2020.02.012
[4]	WANG K B, LIU F, LIU Z H, et al. Analysis of chemical components in oolong tea in relation to perceived quality[J]. International Journal of Food Science & Technology,2010,45(5):913−920.
[5]	LEI S C, XIE M H, HU B, et al. Effective synthesis of theaflavin-3, 3′-digallate with epigallocatechin-3-O-gallate and epicatechin gallate as substrates by using immobilized pear polyphenol oxidase[J]. International Journal of Biological Macromolecules,2017,94:709−718. doi: 10.1016/j.ijbiomac.2016.10.072
[6]	LI Z Y, FENG C X, LUO X G, et al. Revealing the influence of microbiota on the quality of Pu-erh tea during fermentation process by shotgun metagenomic and metabolomic analysis[J]. Food Microbiology,2018,76:405−415. doi: 10.1016/j.fm.2018.07.001
[7]	CHENG L Z, WANG Y F, ZHANG J R, et al. Dynamic changes of metabolic profile and taste quality during the long-term aging of Qingzhuan tea: The impact of storage age[J]. Food Chemistry,2021,359(15):129953. doi: 10.1016/j.foodchem.2021.129953
[8]	CHOI J Y, KIM E M, KO B J, et al. Production of theasinensin A using laccase as antioxidant and antiaging agent[J]. Biotechnology and Bioprocess Engineering,2022,27(2):253−261. doi: 10.1007/s12257-021-0145-7
[9]	TANAKA T, WATARUMI S, MATSUO Y, et al. Production of theasinensins A and D, epigallocatechin gallate dimers of black tea, by oxidation-reduction dismutation of dehydrotheasinensin A[J]. Tetrahedron,2003,59(40):7939−7947. doi: 10.1016/j.tet.2003.08.025
[10]	ZHANG H, QI R L, MINE Y. The impact of oolong and black tea polyphenols on human health[J]. Food Bioscience,2019,29:55−61. doi: 10.1016/j.fbio.2019.03.009
[11]	刘洪林. 工夫红茶咖啡碱和5种儿茶素组分近红外快速测定方法的研究[J]. 食品工业科技,2016,37(15):316−320. [LIU H L. Research of rapid measurement methods of caffeine and five kinds of catechin components quality ingredients of congou black tea using near infrared spectroscopy[J]. Science and Technology of Food Industry,2016,37(15):316−320. LIU H L. Research of rapid measurement methods of caffeine and five kinds of catechin components quality ingredients of congou black tea using near infrared spectroscopy[J]. Science and Technology of Food Industry, 2016, 37(15): 316-320.
[12]	WANG K B, CHEN Q C, LIN Y, et al. Separation of catechins and O-methylated (-)-epigallocatechin gallate using polyamide thin-layer chromatography[J]. Journal of Chromatography B,2016,1017−1018:221−225. doi: 10.1016/j.jchromb.2015.11.060
[13]	NEILSON A P, GREEN R J, WOOD K V, et al. High-throughput analysis of catechins and theaflavins by high performance liquid chromatography with diode array detection[J]. Journal of Chromatography A,2006,1132(1-2):132−140. doi: 10.1016/j.chroma.2006.07.059
[14]	ZHANG J Y, CUI H C, JIANG H Y, et al. Rapid determination of theaflavins by HPLC with a new monolithic column[J]. Czech Journal of Food Sciences,2019,37(2):112−119. doi: 10.17221/213/2018-CJFS
[15]	WEI K, WANG L Y, ZHOU J, et al. Comparison of catechins and purine alkaloids in albino and normal green tea cultivars (Camellia sinensis L.) by HPLC[J]. Food Chemistry,2012,130(3):720−724. doi: 10.1016/j.foodchem.2011.07.092
[16]	GANGULY S, G T K, MANTHA S, et al. Simultaneous determination of black tea-derived catechins and theaflavins in tissues of tea consuming animals using ultra-performance liquid-chromatography tandem mass spectrometry[J]. PLoS One,2016,11(10):e0163498. doi: 10.1371/journal.pone.0163498
[17]	SVOBODA P, VLČKOVÁ H, NOVÁKOVÁ L. Development and validation of UHPLC-MS/MS method for determination of eight naturally occurring catechin derivatives in various tea samples and the role of matrix effects[J]. Journal of Pharmaceutical and Biomedical Analysis,2015,114(10):62−70. doi: 10.1016/j.jpba.2015.04.026
[18]	TAO W Q, ZHOU Z G, ZHAO B, et al. Simultaneous determination of eight catechins and four theaflavins in green, black and oolong tea using new HPLC-MS-MS method[J]. Journal of Pharmaceutical and Biomedical Analysis,2016,131(30):140−145. doi: 10.1016/j.jpba.2016.08.020
[19]	WANG L, YAN T, ZHANG K X, et al. A sensitive UPLC-MS/MS method for simultaneous determination of polyphenols and theaflavins in rat plasma: Application to a pharmacokinetic study of Da Hong Pao tea[J]. Biomedical Chromatography,2019,33(4):e4470. doi: 10.1002/bmc.4470
[20]	TANAKA T, WATARUMI S, FUJIEDA M, et al. New black tea polyphenol having N-ethyl-2-pyrrolidinone moiety derived from tea amino acid theanine: Isolation, characterization and partial synthesis[J]. Food Chemistry,2005,93(1):81−87. doi: 10.1016/j.foodchem.2004.09.013
[21]	TAO S N, CHEN G J, XU W Q, et al. Preparation of theasinensin A and theasinensin B and exploration of their inhibitory mechanism on α-glucosidase[J]. Food Function,2020,11(4):3527−3538. doi: 10.1039/C9FO03054A
[22]	薛金金, 江和源, 龙丹, 等. HPLC法同时测定茶叶中聚酯型儿茶素和茶黄素[J]. 中国食品学报,2014,14(5):237−243. [XUE J J, JIANG H Y, LONG D, et al. Simultaneous determination of theasinensins and theaflavins content in tea by HPLC[J]. Journal of Chinese Institute of Food Science and Technology,2014,14(5):237−243. XUE J J, JIANG H Y, LONG D, et al. Simultaneous determination of theasinensins and theaflavins content in tea by HPLC[J]. Journal of Chinese Institute of Food Science and Technology, 2014, 14(5): 237-243.
[23]	AN T T, CHEN M X, ZU Z Q, et al. Untargeted and targeted metabolomics reveal changes in the chemical constituents of instant dark tea during liquid-state fermentation by Eurotium cristatum[J]. Food Research International,2021,148:110623. doi: 10.1016/j.foodres.2021.110623
[24]	MATSUO Y, TANAKA T, KOUNO I. A new mechanism for oxidation of epigallocatechin and production of benzotropolone pigments[J]. Tetrahedron,2006,62(20):4774−4783. doi: 10.1016/j.tet.2006.03.021
[25]	XIAO Y, LI M Y, LIU Y, et al. The effect of Eurotium cristatum (MF800948) fermentation on the quality of autumn green tea[J]. Food Chemistry,2021,358(1):129848. doi: 10.1016/j.foodchem.2021.129848
[26]	ZHU M Z, LI N, ZHOU F, et al. Microbial bioconversion of the chemical components in dark tea[J]. Food Chemistry,2020,312(15):126043. doi: 10.1016/j.foodchem.2019.126043
[27]	XUE J J, LIU P P, GUO G Y, et al. Profiling of dynamic changes in non-volatile metabolites of shaken black tea during the manufacturing process using targeted and non-targeted metabolomics analysis[J]. LWT-Food Science and Technology,2022,156(15):113010. doi: 10.1016/j.lwt.2021.113010
[28]	王伟伟, 乐婷, 杨刘艳, 等. 骏眉工艺红茶化学成分及其抗氧化活性研究[J]. 食品安全质量检测学报,2022,13(10):3320−3327. [WANG W W, LE T, YANG L Y, et al. Study on the chemical constituents and antioxidant activity of black tea with Junmei technology[J]. Journal of Food Safety and Quality Inspection,2022,13(10):3320−3327. WANG W W, LE T, YANG L Y, et al. Study on the chemical constituents and antioxidant activity of black tea with Junmei technology[J]. Journal of Food Safety and Quality Inspection, 2022, 13(10): 3320-3327.
[29]	HUANG Y Y, XIAO X D, CONG L, et al. A fermented tea with high levels of gallic acid processed by anaerobic solid-state fermentation[J]. LWT - Food Science and Technology,2016,71:260−267. doi: 10.1016/j.lwt.2016.03.047
[30]	MARUYAMA K, KIHARA-NEGISHI F, OHKURA N, et al. Simultaneous determination of catechins and caffeine in green tea-based beverages and foods for specified health uses[J]. Food and Nutrition Sciences,2017,8(3):316−325. doi: 10.4236/fns.2017.83021
[31]	LIU L, SHI J J, YUAN Y H, et al. Changes in the metabolite composition and enzyme activity of fermented tea during processing[J]. Food Research International,2022,158:111428. doi: 10.1016/j.foodres.2022.111428