Full Validation of a Simple Method for Determination of Catechins and Caffeine in Brazilian Green Tea (Camellia sinensis var. assamica) Using HPLC

摘要

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.