The Comparative Qualitative and Quantative Analysis on Green and Black Tea (Camellia sinensis) Extracted by Three Solvents Studied in Sudan
By
Murwan K. Sabah EL-Kheir 1
1. Department of Biochemistry, School of Biotechnology, Faculty of Science and
Technology, El Neelain University, P.O.Box 12702, Khartoum, Sudan
Email address; murwansabahelkhier@yahoo.com
Abstract: The target of this investigate is to investigate the phytochemicals shade of Green and Black Tea extracted by water, ethanol and inorganic substance ether. The results suggested that the participation of alkaloid, tannin, steroid, flavonoid and carbohydrate in Green and Black Tea extracted H2O ethanol and inorganic substance ether. In addition, Green and Black Tea extracted by H2O contained saponins, but Green and Black Tea extracted by ethanol and inorganic substance sky do not contained saponins.Green and Black Tea extracted by ethanol and inorganic substance sky contained cardenolide, but Green and Black Tea extracted by H2O do not contained cardenolide and at last Green and Black Tea extracted by H2O ethanol and inorganic substance sky do not enclose anthraquiaone and coumarin. The alkaloid calm of Green Tea extracted by water, ethanol and inorganic substance sky is 5.4, 4.3 and 4.6 Mg/g, respectively. While alkaloid calm of Black Tea extracted by water, ethanol and inorganic substance sky is 5.4, 4.3, and 4.Mg/g, respectively. The tannin essence of Green Tea extracted by water, ethanol and inorganic substance sky are 1.2, 1.8 and 1.5 %, respectively. While tannin essence of Black Tea extracted by water, ethanol and inorganic substance sky are 1.2, 0.5, 1 %, respectively.
Keywords: Tea, Alkaloid, Tannin and Qualitative.
1.0 Introduction
Green and Black Tea is made up from Camellia sinensis, that is characterized as a lasting evergreen shrub. Tea varieties reflected the flourishing zone (Ceylon and Assam), districted (Darjeeling), form (pekoe and gunpowder) and estimate way (green, black and oolong). India and Srilanka is leading producers of immature tea. The Turkish trader's reportedly introduced to Western enlightenment in 6th Century (1) .It has many uses for its gentle opiate and therapeutic properties (prevent cancer (2), prostate cancer (3), asthma (4), dental cavity, diabetes (5), heart assault (6), high cholesterol (7), upgrade flood (3), mental recall enhancement, mental opening and menopausal symptoms). The immature tea is ready by exposing the collected leaves to air until remaining dampness is eliminated, leaves roasted over a section timber glow and ceaselessly influenced until leaves turn wet and flaccid, after that leaves pass to rolling list and rolled in to balls and subjected to pressure that twist them and gets absolved of the moisture, then leaves jolt out on prosaic trays, once again leaves roasted over a slow and solid colourless glow and kept leaves in hurried suit for an hour to hour and half., compartment leaves pretence a dullish immature colour after that the leaves winnowed, screened, and graded in to not similar types. While black tea, collected leaves are unprotected to air for long period, collected up, tossed until soothing and flaccid, serve exposure, roasted in an iron vessel for about 5 mins and after rolling and dire the leaves are jarred out, unprotected to the externa; air for a few hours, re-roasted for 3 or 4 mins and recoiled, expansion out in baskets and unprotected to feverishness of colourless glow for 5 or 6 mins and then rolled for third time and once again feverishness and at last dusty in baskets over colourless glow from that leaves turn black in colour. Steroid compounds are containing the perhydrocyclopentano-phenanthrene structure of the body and often happen in glycosidal multiple with sugarine (8). Alkaloids are organisation of innate products (atropine, quinine, hypnotic ...etc) that are at large used to provide Malaria and Cancer diseases (8). Cardenolides are deed on the heart send or surreptitious resource to complement the force and quickness of the contraction, thus it is well known as cardiotonic (8). Carbohydrates are the many bountiful organic voters of plant, serve as the leading source of containing alkali appetite (sugar and starch), together with critical voters of ancillary hankie (cellulose in wood, string and flax) (8). Anthraquinones are oxygenated derivative of pharmacological significance that is used as laxatives or cathartics, anti-inflammatory, antibacterial, antifungal and moreover as innate dyes (8). The flavonoid, consecrate one of many heterogeneous and extensive organisation of innate products, found in fruit, vegetables, nuts, seeds, stem, flowers, together with tea, and are one of the many critical voters of the human diet(8). Coumarins are derivative of benz-? pyrone, or lactones of o-hydroxcinnamic poison such as coumarin, umbelliferonee, aesculetin, daphnetin, fraxetin and scopoletin, they are found in both state in giveaway or multiple with glycosidic., not all members of the organisation are phenolic (8).The tannin compounds are phenolic polymers curt protein from aqueous answer and it is lower or restrain chemical substance wake up (9).
Objectives of this investigate are Phytochemicals screening of the immature and black tea, that included:(1) Qualitative review of Alkaloid, Anthraquiaones, Cardenolides, Tannin, Saponins, Coumarin, Flavonoid, Steroid and Carbohydrates in immature and black tea extracted by water, ethanol and inorganic substance sky in immature and black tea extracted by water, ethanol and inorganic substance ether. (2) Quantative review of Alkaloid and Tannin calm
2.0 Materials and methods
2.1 Sample preparation: The dusty immature and black teas were collected from local marketplace of Khartoum State, Sudan., the dusty teas (green and black) were liberated from unfamiliar materials and belligerent in to excellent powder to pass 0.4 mm netting screen. The ready samples were kept in parsimonious containers and stored at room temperature until analysis. Preparation of e aqueous descent of tea (green and black) by using water, ethanol and inorganic substance sky is carried out according to way described by (10).
2.2 Qualitative analysis: Alkaloid, Saponins, Tannin, Steroid, Flavonoids, Comrine, Cardiac and Carbohydrates were identified according to way described by (10).
2.2.1 Alkaloid: Two gram of well belligerent samples was extracted with 10 ml of 1%HCL for 30 mins in H2O bath. The postponement was filtrated by string whool in to blood vessel and the secrete answer was widely separated in to two portions A and B. Then jam-packed answer of sodium bicarbonate was updated to the secrete answer (B) compartment pH turn 8 -9 and the answer was churned with 3 ml chloroform, then top covering was private by pipette and treated with colour with acetic poison compartment pH turn 5 and 5 drops of dragendroff,s monarch were added. If the curt is form. This indicated the participation of quaternary alkaloid. The lower covering (chloroform) was treated with colour with 3 ml of 1%HCL that separated the lower covering in to two layers. Then top covering was pipette and treated with colour with dragendroff,s regent, this indicated to participation of tertiary alkaloid.
2.2.2 Anthraquinones: 0.5 gram of well belligerent representation was extracted in hot H2O for two mins with 5 ml 0.5N KOH and 0.5 ml H2O. Suspension was cooled and secrete by using potion nap and 6 drops of acetic poison was added, then the answer was churned with 5 ml benzene, The top covering was separated and pipette in to assessment blood vessel followed by adding of 2 ml 0.0N KOH. If the answer gives red colour. This indicated the participation of Anthraquinones.
2.2.3 Cardenolides: 3 gram of well belligerent representation was extracted 2 ml of press out H2O in hot H2O bath for 30 minutes, then enable the answer to cold and secrete by using potion nap in to assessment tube. 10 drops of lead acetate was updated and then secrete to blood vessel (A) and (B). Four drops of Keddle,s reagent was updated The answer (B) was churned with 2 ml chloroform, and 4 drops of Keddle,s reagent was updated to lower layer. If the answer gives a violet colour. This indicated the participation of Cardenolides.
2.2.4 Tannin: 3 gram of well belligerent representation was extracted with 1 ml press out H2O in hot H2O bath for 5 minutes, then secrete the answer by using filter paper and enable a answer to cold . The filtration answer was widely separated in to two portions A and B. 5 ml of 2% NaCL was updated to part A. Suspended answer was filtrated by using filter paper, followed by adding 5 ml 1% gelatin solution. Precipitate indicated the participation of Tannin. 1 ml of 1% ferric chloride was updated to part B. The immature blue colour indicated the participation of Tannin.
2.2.5 Saponins: 3 gram of well belligerent representation was extracted with 5 ml press out H2O in hot H2O bath for 6 minutes. Then secrete the answer by using potion wool. 1 ml of representation extracted was jarred for 5 mins and at last the persistent and voluminous stew is formed.
2.2.6 Coumarin: 3 gram of well belligerent representation was extracted with 5 ml press out H2O in hot H2O bath for 6 minutes, then secrete the answer by using filter paper and enable a answer to cool. Then the representation extracted shows a blue fluorescence, but it altered in to greenish - blue on add-on of few drops of ammonia. This indicated to participation of coumarin.
2.2.7 Flavonoid: 3 gram of well belligerent representation was extracted 2 ml of press out H2O in hot H2O bath for 30 minutes, then enable the answer to cold and secrete by using potion nap in to assessment tube. 10 drops of lead acetate was updated to filtration. The yellowish curt is formed. This indicated to participation of Flavonoid.
2.2.8 Steroid: 3 gram of well belligerent representation was extracted with 5 ml ethanol with 1% acetic poison in hot H2O bath for 5 minutes, then secrete the answer by using filter paper and enable a answer to cool. The filtration answer was widely separated in to two portions A and B. Few drops of SbCL3 was updated to part A and few drops of Marquis's reagent was updated to part B. red colour rise in part A and violet to red colour rise in part B. This indicated to participation of Steroid.
2.2.9 Carbohydrates: 3 gram of well belligerent representation was extracted with 5 ml press out H2O in hot H2O bath for 6 minutes. Then filtration the answer by using potion wool. Few drops of Molish, s reagent were updated to filtration solution. Then violet colour appeared, this indicated to participation of Carbohydrates.
2.3 Quantative analysis:
2.3.1 Alkaloid content: It is gritty according to way described by (8).
Evaporated amalgamated remove on steam bath with air stream to about 10 ml increase deliberate surplus 0.2N H2SO4 and go on evaporation to remove the solvent, cold and increase methyl. Then the surplus poison (H2SO4) was titrated with 0.02 N NaOH.
V1 X M1 = V2 X M2 (1)
Mg/g = V2 X D.F. X M.WT (2)
103 X S
Where: V1 = Volume of 0.20.2N H2SO4, M1 = Concentration of H2SO4 (0.2N), V2 = Volume of NaOH, M2 = Concentration of NaOH (0.02 N), D.F. = Dilution factor, M.WT = Molecular weight of NaOH, S= Weight of representation and 103 = conversion reason from volume to weight.
2.3.2 Tannin content: It is gritty according to way described by (11).
0.2 gram of extracted representation was placed in 50 ml of conical flask and 10 ml of 1% HCL was added. The essence of flask were jarred for 20 minutes, and then essence of the flask were centrifuged at 3000 rpm for 5 minutes. Then essence of flask were incubated at 30 oC for 20 minutes. The spectrophotometer was read at 500 nm and then scale read 0 with the vacant solution. Finally, readings were taken 3 times and averaged.
- C.E. (%) = C X 100 (1)
10 X S
Where: C.E. = Catechin equivalent, C = Concentration analogous to visual density, 10 = Volume of remove (ml) and S = Weight of sample.
3.0 Results and Discussion
3.1 Qualitative analysis: The qualitative assessment of immature and black tea that extracted by water, ethanol and inorganic substance sky is to alkaloid, anthraquiaones, saponins, tannin, cardenolides, steroid, flavonoids, coumrine, and carbohydrates were shown in Table (1). The commentary suggested that the alkaloid, tannin, steroid, flavanoid in 3 extractions (water, ethanol and inorganic substance ether) of immature and black tea are available. The cardenolide in two extractions (ethanol and inorganic substance ether) of immature and black tea are participation but the cardenolide in H2O descent is not found in immature and black tea. These results indicated that cardenolide is not dissolved in H2O but dissolved in ethanol and inorganic substance ether. In add-on to that the anthraquiaones and coumarin 3 extractions (water, ethanol and inorganic substance ether) of immature and black tea are not available. The saponin in immature and black tea extracted by H2O is present, but saponin in immature and black tea extracted by ethanol and inorganic substance sky is not present These commentary suggested that immature and black tea do not enclose anthraquiaones and coumarin.These results are upheld by those results since by (12) ..
3.2 Quantative analysis: Table (2) indicated that the alkaloid calm in immature tea in water, ethanol and inorganic substance extracted is 5.4, 4.3 and 4.6%, respectively. While the alkaloid calm in black tea in water, ethanol and inorganic substance extracted is 6.4, 4.5 and 5.2 %, respectively. The results suggested that alkaloid in both immature and black tea in H2O extracted is aloft than in ethanol and inorganic substance sky extracted, but there is no poignant disparity in 3 solvents extracted for immature tea at (P ? 0.05).This indicated that descent of alkaloid is not effected by variety of solvents. In add-on to that the results indicated that tannin calm in immature and black tea for H2O extracted is similar, tannin calm in immature tea for ethanol extracted is aloft than in black tea. The results suggested there poignant disparity at (P ? 0.05). The movement in alkaloid and tannin calm is attributed to sort of solvent, sort of tea and routine of tea.
Conclusion and Recommendation:
- We resolved that the immature and black tea enclose alkaloid, tannin, steroid, flavonoid, cardenolides, saponins and carbohydrate. In add-on amount of alkaloid and tannin are sundry according to sort of well-off used. Our recommendation, the over investigate in immature and black tea should be completed quite antimicrobial study.
References
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Table (1): Chemical voters that existing ( ve) and non-available (- ve) in immature and black tea.
Solvent descent /
Compounds
Water extraction
Ethanol extraction
Petroleum sky extraction
Green tea
Black tea
Green tea
Black tea
Green tea
Black tea
1/Alkaloid
( ) ve
( ) ve
( ) ve
( ) ve
( ) ve
( ) ve
2/Anthraquinones
(-) ve
(-) ve
(-) ve
(-) ve
(-) ve
(-) ve
3/Cardenolides
(-) ve
(-) ve
( ) ve
( ) ve
( ) ve
( ) ve
4/Tannin
( ) ve
( ) ve
( ) ve
( ) ve
( ) ve
( ) ve
5/Saponins
( ) ve
( ) ve
(-) ve
(-) ve
(-) ve
(-) ve
6/Coumarin
(-) ve
(-) ve
(-) ve
(-) ve
(-) ve
(-) ve
7/Flavonoids
( ) ve
( ) ve
( ) ve
( ) ve
( ) ve
( ) ve
8/Steroid
( ) ve
( ) ve
( ) ve
( ) ve
( ) ve
( ) ve
9/Carbohydrates
( ) ve
( ) ve
( ) ve
( ) ve
( ) ve
( ) ve
Table (2): Alkaloid and tannin calm in extracted " tea well-off (water, ethanol and inorganic substance sky of immature and black type.
Type of solvent
Water extraction
Ethanol extraction
Petroleum sky extraction
Type of tea
Green tea
Black tea
Green tea
Black tea
Green tea
Black tea
Alkaloid Mg/g
5.4a1
6.4 a 0.5
4.3 a 0.3
4.5 a 1.2
4.6 a 1
5.2 a 1.4
Tannin %
1.2 a 0.5
1.2 a 1
1.8 a 2
0.5 b 0.01
1.5 a 1
1.0 a 0.8
* Each worth is median of 3 replicates voiced on dehydrated weight basis.
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