Qualitative analysis of catechins from green tea GMB-4 clone using HPLC and LC-MS/MS

ObjectiveTo identify the bioactive compounds in catechins isolation and its components from green tea GMB-4 clone.MethodsGreen tea GMB-4 clones were extracted with distilled water at 90 °C. Samples were eluted into the column with 10% ethanol. Subsequently, the column was eluted with 95% ethanol and evaporated separately. Green tea extract was identified by thin layer chromatography. Catechins were separated by the stationary phase in column chromatography using polyamide with 10% ethanol eluent and 95% ethanol. The results of isolations were analyzed by high performance liquid chromatographic (HPLC) and LC-MS/MS. Analysis of catechins by HPLC was done by external standard.ResultsFraction from 10% ethanol showed that four major peaks at retention time of 1.663, 2.367, 2.950 and 4.890, indicated the presence of four catechins components including catechin, epicatechins, gallocatechin and epigallocatechin. Whereas, fraction from 95% ethanol showed two main peaks at retention time of 5.167 and 9.82, which indicated the presence of epigallocatechin gallate (EGCG) and epicatechin gallate (ECG). EGCG (m/z 459), epigallocatechin (m/z 307), ECG (m/z 443), and epicatechin (m/z 291) were isolated and separated successfully using HPLC and LC-MS/MS.ConclusionsThe HPLC and LC-MS/MS methods were successfully tuned for the qualitative analysis of green tea extract with EGCG and ECG. Four major catechins were separated and identified by LC-MS/MS, such as EGCG, epigallocatechin, ECG and epicatechin. The result of HPLC analysis showed that EGCG and ECG were main components from catechins isolation of green tea GMB-4 clone.