Microwave-assisted extraction of tea polyphenols and tea caffeine
from green tea leaves
PRESENTED BY: VIKAS SHARMA [Link].
(PHARMACOGNOSY)
Introduction
Green tea leaves contain tea polyphenols, tea caffeine, amino acids, saponins, tannins, etc., with about 10-30% (w/w) polyphenols and 2-4% (w/w) caffeine. Tea polyphenols include catechines, flavanols, flavanones, phenolic acids, glycosides and the aglycones of plant pigments. They are soluble in water, ethanol, methanol, acetone etc.
Tea polyphenols isolated from green tea leaves are natural antioxidant [1] and have a scavenging effect on active oxygen radical [2].
Tea polyphenols have a stronger anti-oxidative activity than Butylated hydroxyanisole, butylated hydroxytoluene and DL-a- tocopherol; and the toxicity of tea polyphenols is lower
than butylated hydroxyanisole, butylated hydroxytoluene
and DL-atocopherol [3]. Microwave digestion of matrices for their eventual elemental analysis has been routinely used for several years [4]. Recently, microwave-assisted extraction (MAE) has been
used for the extraction of biologically active compounds,
such as extraction of essential oils from the leaves of rosemary and peppermint [5],
Objectives :
The purpose of this work was to develop a MAE method and evaluate MAE and conventional extraction methods for the
extraction of tea polyphenols and tea caffeine from green tea
leaves.
Materials and methods
2.1. Plant materials and chemicals
Whole green tea leaves (Thea sinensis L.) (producing area:
Hangzhou, China) were purchased from a local market. Ethanol, methanol and acetone used in the experimental work were all of analytical reagent grade chemicals. Caffeine and all other chemicals for analysis of tea polyphenols and caffeine used were all of analytical reagent grade chemicals.
2.2. Microwave-assisted extraction A household microwave oven (full power 700 W, National, Japan) was modified in our laboratory [12]. Green tea leaves were mixed with an appropriate solvent. The suspensions were irradiated with microwaves as follows: 45 s power on (heating to the desired temperature about 85 90C ) and 10 s power off and then 3 s power on (for heating) and 10 s power off (for cooling) and so on to the pre-set extraction time. Super boiling of the solution did not occur. There is no microwave energy radiation by magnetic stirring for 0- 90 min pre-leaching at room temperature and then MAE for 4 min.
2.3. Colorimetric analysis of tea polyphenols [15]
1ml extract, 4 ml water and 5 ml ferrous tartrate tetrahydrate solution were mixed with 15 ml buffer solution (0.067 mol /l pH 7.5 potassium phosphate) in 25-ml capacity bottle. It takes several minutes for color to develop. With blank solution (solvent) as reference solution, absorbance (A) at Vis 540 nm with a 10 mm quartz cell was used to calculate the extraction of tea polyphenols according to Eq. (1) [15]. The errors were controlled to less than 0.5% through duplicated experiments and analysis.
Eq. (1)
where, L1, total volume of extract, ml; L2 volume of extract
used for colorimetric analysis, ml; M, mass of tea leaves, mg;
A, absorbance at Vis 540 nm; 1.957, when absorbance was 0.5 at Vis 540 nm with a 10 mm quartz cell, the concentration of tea polyphenols was 1.957 mg /ml
2.4. Colorimetric analysis of tea caffeine [16]
20 ml of extract, 10 ml hydrochloric acid (0.01 mol/l ) and 2 ml lead acetate basic solution were mixed with 218 ml water in a 250-ml volumetric flask. The mixed solution was stand for 1 h and then was filtered. After that, 50 ml filtered solution and 0.2 ml H2SO4 solution (4.5 mol/l) were mixed with 49.8 ml water in a 100-ml volumetric flask. The mixed solution was stand for 30 min and then was filtered.
With blank solution (solvent) as reference solution, the filtered solution was measured at UV 274 nm with a 10 mm quartz cell.
Eq. (2) where C, the concentration of caffeine calculated from the calibrated regression equation based on the absorbance at UV 274 nm, mg /ml ; L1 , total volume of extract, ml; M, mass of green tea leaves, mg
In the present work, the percentage of tea polyphenols or tea
caffeine in extracts was defined as following Eq.(3): The percentage of tea polyphenols or tea caffeine in extracts (%; w=w)
X 100 %
(Eq. 3)
3. Results and discussion
3.1. The effect of ethanol concentration on the extraction of polyphenols and caffeine
3.2. The effect of MAE time on the extraction of polyphenols and caffeine
3.3. The effect of liquid/solid ratio on the extraction of polyphenols and caffeine
3.4. The effect of different solvents on the extraction of polyphenols and caffeine
Fig. 4. (a) and(b)The effect of solvents used on the extraction of polyphenols and caffeine Solvent: 100 ml; whole green tea leaves: 5 g; MAE for 4min; liquid/solid ratio: 20:1 ml /g
.
3.5. The effect of pre-leaching time before MAE on the extraction of polyphenols and caffeine
3.6. Comparison of MAE and conventional extraction methods
4. Conclusion
MAE has been shown to be an efficient method for extraction of polyphenols and caffeine from green tea leaves.
Compared with the conventional extraction methods, the
MAE procedure provided high extraction, high extraction selectivity, requiring short time, and less labor intensive.
MAE was suitable for fast extraction of polyphenols and
caffeine from green tea leaves. Food and medicinal industries would be benefited from this emerging technology of MAE , which was rapid, safe and more ecofriendly than conventional extraction methods.
References
