Full Validation of a Simple Method for Determination of Catechins and Caffeine in Brazilian Green Tea (Camellia sinensis var. assamica) Using HPLC |
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Authors: | S. T. Saito P. E. Fröehlich G. Gosmann A. M. Bergold |
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Affiliation: | 1. Programa de Pós-Gradua??o em Ciências Farmacêuticas, Faculdade de Farmácia, Universidade Federal do Rio Grande do Sul (UFRGS), Av. Ipiranga 2752, Porto Alegre, 90610-000, RS, Brazil
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Abstract: | This paper describes the validation of an HPLC method for the assay of a green tea brew. The method employs a RP-18 column with water:methanol:ethyl acetate elution and UV detection at 280 nm. Specificity was evaluated using a photodiode array detector. The validation data showed that the assay is specific, accurate, precise, and reproducible for determination of six catechins and caffeine simultaneously. The response was linear over a range of 37–185 μg mL?1 for caffeine, 99–500 μg mL?1 for (?)-epigallocatechin (EGC), 20–100 μg mL?1 for (+)-catechin (C), 30–150 μg mL?1 for (?)-epicatechin (EC), 150–800 μg mL?1 for (?)-epigallocatechin gallate (EGCG), 20–105 μg mL?1 for (?)-gallocatechin gallate (GCG) and 40–205 μg mL?1 for (?)-epicatechin gallate (ECG) (r > 0.9999 for all compounds). The range of recoveries was 96.12–110.48% according to substances. The RSD values for intra- and inter-day precision studies were <2.07 and <6.65%, respectively. The composition of samples assayed suggests that the summer is the best season for extract a major content of EGCG and caffeine. This assay can be readily utilized as quality controlled method for major green tea compounds. |
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