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Efficient procedure for isolating methylated catechins from green tea and effective simultaneous analysis of ten catechins, three purine alkaloids, and gallic acid in tea by high-performance liquid chromatography with diode array detection.
J Chromatogr A. 2009 Apr 10; 1216(15):3223-31.JC

Abstract

Monomers of (-)-epigallocatechin (EGC), (-)-epigallocatechin gallate (EGCG), (-)-epicatechin (EC), (-)-epicatechin gallate (ECG), (-)-epigallocatechin 3-O-(3-O-methyl) gallate (EGCG3''Me) and (-)-3-O-methyl epicatechin gallate (ECG3'Me) (purity, >97%) were successfully prepared from extract of green tea by two-time separation with Toyopearl HW-40S column chromatography eluted by 80% ethanol. In addition, monomers of (-)-catechin (C), (-)-gallocatechin (GC), (-)-gallocatechin gallate (GCG), and (-)-catechin gallate (CG) (purity, >98%) were prepared from EC, EGC, EGCG, and ECG by heat-epimerization and semi-preparative HPLC chromatography. With the prepared catechin standards, an effective and simultaneous HPLC method for the analysis of gallic acid, tea catechins, and purine alkaloids in tea was developed in the present study. Using an ODS-100Z C(18) reversed-phase column, fourteen compounds were rapidly separated within 15min by a linear gradient elution of formic acid solution (pH 2.5) and methanol. A 2.5-7-fold reduction in HPLC analysis time was obtained from existing analytical methods (40-105min) for gallic acid, tea catechins including O-methylated catechins and epimers of epicatechins, as well as purine alkaloids. Detection limits were generally on the order of 0.1-1.0ng for most components at the applied wavelength of 280nm. Method replication generally resulted in intraday and interday peak area variation of <6% for most tested components in green, Oolong, black, and pu-erh teas. Recovery rates were generally within the range of 92-106% with RSDs less than 4.39%. Therefore, advancement has been readily achievable with commonly used chromatography equipments in the present study, which will facilitate the analytical, clinical, and other studies of tea catechins.

Authors+Show Affiliations

College of Food Science and Technology, Nanjing Agricultural University, Nanjing, China.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

19246045

Citation

Hu, Bing, et al. "Efficient Procedure for Isolating Methylated Catechins From Green Tea and Effective Simultaneous Analysis of Ten Catechins, Three Purine Alkaloids, and Gallic Acid in Tea By High-performance Liquid Chromatography With Diode Array Detection." Journal of Chromatography. A, vol. 1216, no. 15, 2009, pp. 3223-31.
Hu B, Wang L, Zhou B, et al. Efficient procedure for isolating methylated catechins from green tea and effective simultaneous analysis of ten catechins, three purine alkaloids, and gallic acid in tea by high-performance liquid chromatography with diode array detection. J Chromatogr A. 2009;1216(15):3223-31.
Hu, B., Wang, L., Zhou, B., Zhang, X., Sun, Y., Ye, H., Zhao, L., Hu, Q., Wang, G., & Zeng, X. (2009). Efficient procedure for isolating methylated catechins from green tea and effective simultaneous analysis of ten catechins, three purine alkaloids, and gallic acid in tea by high-performance liquid chromatography with diode array detection. Journal of Chromatography. A, 1216(15), 3223-31. https://doi.org/10.1016/j.chroma.2009.02.020
Hu B, et al. Efficient Procedure for Isolating Methylated Catechins From Green Tea and Effective Simultaneous Analysis of Ten Catechins, Three Purine Alkaloids, and Gallic Acid in Tea By High-performance Liquid Chromatography With Diode Array Detection. J Chromatogr A. 2009 Apr 10;1216(15):3223-31. PubMed PMID: 19246045.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Efficient procedure for isolating methylated catechins from green tea and effective simultaneous analysis of ten catechins, three purine alkaloids, and gallic acid in tea by high-performance liquid chromatography with diode array detection. AU - Hu,Bing, AU - Wang,Lin, AU - Zhou,Bei, AU - Zhang,Xin, AU - Sun,Yi, AU - Ye,Hong, AU - Zhao,Liyan, AU - Hu,Qiuhui, AU - Wang,Guoxiang, AU - Zeng,Xiaoxiong, Y1 - 2009/02/13/ PY - 2008/10/30/received PY - 2009/02/03/revised PY - 2009/02/09/accepted PY - 2009/2/28/entrez PY - 2009/2/28/pubmed PY - 2009/9/11/medline SP - 3223 EP - 31 JF - Journal of chromatography. A JO - J Chromatogr A VL - 1216 IS - 15 N2 - Monomers of (-)-epigallocatechin (EGC), (-)-epigallocatechin gallate (EGCG), (-)-epicatechin (EC), (-)-epicatechin gallate (ECG), (-)-epigallocatechin 3-O-(3-O-methyl) gallate (EGCG3''Me) and (-)-3-O-methyl epicatechin gallate (ECG3'Me) (purity, >97%) were successfully prepared from extract of green tea by two-time separation with Toyopearl HW-40S column chromatography eluted by 80% ethanol. In addition, monomers of (-)-catechin (C), (-)-gallocatechin (GC), (-)-gallocatechin gallate (GCG), and (-)-catechin gallate (CG) (purity, >98%) were prepared from EC, EGC, EGCG, and ECG by heat-epimerization and semi-preparative HPLC chromatography. With the prepared catechin standards, an effective and simultaneous HPLC method for the analysis of gallic acid, tea catechins, and purine alkaloids in tea was developed in the present study. Using an ODS-100Z C(18) reversed-phase column, fourteen compounds were rapidly separated within 15min by a linear gradient elution of formic acid solution (pH 2.5) and methanol. A 2.5-7-fold reduction in HPLC analysis time was obtained from existing analytical methods (40-105min) for gallic acid, tea catechins including O-methylated catechins and epimers of epicatechins, as well as purine alkaloids. Detection limits were generally on the order of 0.1-1.0ng for most components at the applied wavelength of 280nm. Method replication generally resulted in intraday and interday peak area variation of <6% for most tested components in green, Oolong, black, and pu-erh teas. Recovery rates were generally within the range of 92-106% with RSDs less than 4.39%. Therefore, advancement has been readily achievable with commonly used chromatography equipments in the present study, which will facilitate the analytical, clinical, and other studies of tea catechins. SN - 1873-3778 UR - https://www.unboundmedicine.com/medline/citation/19246045/Efficient_procedure_for_isolating_methylated_catechins_from_green_tea_and_effective_simultaneous_analysis_of_ten_catechins_three_purine_alkaloids_and_gallic_acid_in_tea_by_high_performance_liquid_chromatography_with_diode_array_detection_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0021-9673(09)00227-1 DB - PRIME DP - Unbound Medicine ER -