THEOBROMINE AND THEOPHYLLINE DETERMINATION IN BLACK TEA BY CAPILLARY ZONE ELECTROPHORESIS USING LARGE VOLUME SAMPLE STACKING

Some features of the formation of methylxanthines during growth and development of tea leaves are discussed, and the information about the content of minor methylxanthines, theobromine (TB) and theophylline (TF), in tea products is summarized. The expediency of using TB and TF as marker substances characterizing the quality of tea is substantiated. Possibility and limitations of using electrophoretic methods for TB and TF determination in tea products were investigated. Experimental studies were carried out to develop a method for TB and TF determination in black tea by capillary zone electrophoresis using the procedure of analytes online concentration – large volume sample stacking (LVSS-CZE-UV). Optimized conditions for the analytes separation were using the borate buffer solution (pH = 9.8, 90 mM concentration) and 20 kV voltage in the system. The following conditions for the analytes concentrating were proposed: time for introducing a 40-fold diluted tea extract (1.0 g per 100 ml of water) was 200 s at 50 mbar; the polarity reversal time was 1.5–1.7 min at a voltage of –10 kV. Under the selected conditions, the TB and TF concentration efficiency factors were 28 and 38, respectively. The analysis time did not exceed 22 min. For the developed method, the repeatability, intralaboratory precision, trueness and accuracy of the results of analytes determination in black tea samples were evaluated. Determining TB (0.04–4.0 mg/g concentration range) and TF (0.04–1.0 mg/g concentration range) in tea showed that the relative standard deviation of the results of the analytes determination σ(Δ) did not exceed 7 and 12 %, respectively. Analysis of various samples of black tea using the developed method showed that TB content ranged from 0.14 to 2.32 mg/g and TF content ranged from "< 0.040" to 0.15 mg/g.