The Comparative Qualitative and Quantative Analysis on Lime and Black Tea Camellia sinensis Extracted by 3 Solvents Studied in Sudan By Murwan K. Departmentof Biochemistry, College of Biotechnology, Faculty of Science and Technology, El Neelain University, P. The conclusions revealed that the presence of alkaloid, tannin, steroid, flavonoid and carbohydrate in Lime and Black Tea extracted h2o ethanol and petroleum ether. In addition, Lime and Black Tea extracted by h2o contained saponins, but Lime and Black Tea extracted by ethanol and petroleum ether not ever contained saponins. Green and Black Tea extracted by ethanol and petroleum ether contained cardenolide, but Lime and Black Tea extracted by h2o not ever contained cardenolide and finally Lime and Black Tea extracted by h2o ethanol and petroleum ether not ever contain anthraquiaone and coumarin.
The alkaloid content of Lime Tea extracted by water, ethanol and petroleum ether is 5. 6 Mg or g, respectively. While alkaloid content of Black Tea extracted by water, ethanol and petroleum ether is 5. Mg or g, respectively. The tannin contents of Lime Tea extracted by water, ethanol and petroleum ether are 1.
While tannin contents of Black Tea extracted by water, ethanol and petroleum ether are 1. 5, two %, respectively. Keywords: Tea, Alkaloid, Tannin and Qualitative. 0 Introduction Lime and Black Tea is created up from Camellia sinensis, that is characterized like a perennial evergreen shrub. Tea varieties reflected the growing region Ceylon and Assam, districted Darjeeling, shape pekoe and gunpowder and processing method green, black and oolong.
India and Srilanka is primary producers of lime tea. The Turkish trader's reportedly introduced to Western lifestyle in 6th Century two. It has many uses for its mild stimulant and medicinal properties prevent cancer 2, prostate cancer 3, asthma 4, dental cavity, diabetes 5, heart attack 6, high cholesterol 7, improve fertility 3, memory enhancement, mental performance and menopausal symptoms. The lime tea is prepared by exposing the gathered leaves to space until superfluous moisture is eliminated, leaves roasted over a brick wood fire and continually stirred until leaves grow to moist and flaccid, subsequent to which leaves pass to rolling table and rolled into balls and subjected to compression which twist them and gets rid regarding the moisture, then leaves shaking out on flat trays, repeatedly leaves roasted over a slow and steady charcoal fire and kept leaves in rapid motion for an hour to hour and half. , till leaves assume a dullish lime colour subsequent to that the leaves winnowed, screened, and graded into different types.
While black tea, gathered leaves are exposed to space for long period, gathered up, tossed until soft and flaccid, distant exposure, roasted in an iron pot for about 5 periods and subsequent to rolling and pressing the leaves are shaken out, exposed to the outer space for some hours, re-roasted for 3 or 4 periods and recoiled, spread out in baskets and exposed to heat of charcoal fire for 5 or six periods and then rolled for third time and repeatedly heat and finally dried in baskets over charcoal fire from which leaves grow to black in colour. Steroid compounds are containing the perhydrocyclopentano-phenanthrene skeleton and usually occur in glycosidal combination with sweetener 8. Alkaloids are team of natural products atropine, quinine, morphine etc which are widely used to treat Malaria and Cancer diseases 8. Cardenolides are act on the heart direct or indirect mechanism to enhance the force and velocity regarding the contraction, that is why it is known as cardiotonic 8. Carbohydrates are the greatest abundant organic constituents of plant, deliver as the primary source of chemical life sweetener and starch, as well as important constituents of supporting tissue cellulose in wood, cotton and flax 8.
Anthraquinones are oxygenated derivative of pharmacological importance that is used as laxatives or cathartics, anti-inflammatory, antibacterial, antifungal and also as natural dyes 8. The flavonoid, constitute two of most diverse and widespread team of natural products, located in fruit, vegetables, nuts, seeds, stem, flowers, as well as tea, and are one regarding the greatest important constituents regarding the person diet 8. Coumarins are derivative of benz-? pyrone, or lactones of o-hydroxcinnamic acid for example coumarin, umbelliferonee, aesculetin, daphnetin, fraxetin and scopoletin, they can be located in most state in free or combination with glycosidic. , not all members regarding the team are phenolic 8. The tannin compounds are phenolic polymers precipitate protein from aqueous solution and it is reduce or inhibit enzyme activity 9.
Objectives of this read are Phytochemicals screening regarding the lime and black tea, which included: two Qualitative analysis of Alkaloid, Anthraquiaones,Cardenolides, Tannin, Saponins, Coumarin, Flavonoid, Steroid and Carbohydrates in lime and black tea extracted by water, ethanol and petroleum ether in lime and black tea extracted by water, ethanol and petroleum ether. 3 Quantative analysis of Alkaloid and Tannin content 2. 0 Fabrics and methods 2. 1 Sample preparation: The dried lime and black teas were collected from regional market of Khartoum State, Sudan. , the dried teas lime and black were freed from foreign fabrics and ground into fine powder to pass 0.
The prepared samples were kept in tight containers and stored at room heat until analysis. Preparation of e aqueous extraction of tea lime and black by creating use of water, ethanol and petroleum ether is carried out regarding to method described by 10. 2 Qualitative analysis: Alkaloid, Saponins, Tannin, Steroid, Flavonoids, Comrine, Cardiac and Carbohydrates were identified regarding to method described by 10. 1 Alkaloid: 3 gram of well ground samples was extracted with 10 ml of 1%HCL for 30 periods in h2o bath. The suspension was filtrated through cotton whool into tube and the filtrate solution was divided into 3 portions A and B.
Then saturated solution of sodium bicarbonate was added to the filtrate solution Be till pH grow to 8 -9 and the solution was mixed with 4 ml chloroform, then upper layer was removed by pipette and treated with acetic acid till pH grow to six and 5 drops of dragendroff,s regent were added. If the precipitate is form. This indicated the presence of quaternary alkaloid. The decreased layer chloroform was treated with 4 ml of 1%HCL which separated the decreased layer into 3 layers. Then upper layer was pipette and treated with dragendroff,s regent, this indicated to presence of tertiary alkaloid.
5 gram of well ground sample was extracted in boiling h2o for 3 periods with six ml 0. Suspension was cooled and filtrate by creating use of glass wool and six drops of acetic acid was added, then the solution was mixed with six ml benzene, The upper layer was separated and pipette into test tube followed by adding of 3 ml 0. If the solution sends red colour. This indicated the presence of Anthraquinones. 3 Cardenolides: 4 gram of well ground sample was extracted 3 ml of distil h2o in boiling h2o bath for 30 minutes, then let the solution to cold and filtrate by creating use of glass wool into test tube.
10 drops of lead acetate was added and then filtrate to tube A and B. 4 drops of Keddle,s reagent was added The solution Be was mixed with 3 ml chloroform, and 5 drops of Keddle,s reagent was added to decreased layer. If the solution sends a violet colour. This indicated the presence of Cardenolides. 4 Tannin: 4 gram of well ground sample was extracted with two ml distil h2o in boiling h2o bath for six minutes, then filtrate the solution by creating use of filter cardboard and let a solution to cool.
The filtration solution was divided into 3 portions A and B. six ml of 2% NaCL was added to portion A. Suspended solution was filtrated by creating use of filter paper, followed by adding six ml 1% gelatin solution. Precipitate indicated the presence of Tannin. two ml of 1% ferric chloride was added to portion B.
The lime blue colour indicated the presence of Tannin. 5 Saponins: 4 gram of well ground sample was extracted with six ml distil h2o in boiling h2o bath for seven minutes. Then filtrate the solution by creating use of glass wool. two ml of sample extracted was shaken for six periods and finally the persistent and voluminous froth is formed. 6 Coumarin: 4 gram of well ground sample was extracted with six ml distil h2o in boiling h2o bath for seven minutes, then filtrate the solution by creating use of filter cardboard and let a solution to cool.
Then the sample extracted shows a blue fluorescence, but it changed into greenish - blue on addition of little drops of ammonia. This indicated to presence of coumarin. 7 Flavonoid: 4 gram of well ground sample was extracted 3 ml of distil h2o in boiling h2o bath for 30 minutes, then let the solution to cold and filtrate by creating use of glass wool into test tube. 10 drops of lead acetate was added to filtration. The yellowish precipitate is formed.
This indicated to presence of Flavonoid. 8 Steroid: 4 gram of well ground sample was extracted with six ml ethanol with 1% acetic acid in boiling h2o bath for six minutes, then filtrate the solution by creating use of filter cardboard and let a solution to cool. The filtration solution was divided into 3 portions A and B. Little drops of SbCL3 was added to portion A and little drops of Marquis's reagent was added to portion B. red colour develop in portion A and violet to red colour develop in portion B.
This indicated to presence of Steroid. 9 Carbohydrates: 4 gram of well ground sample was extracted with six ml distil h2o in boiling h2o bath for seven minutes. Then filtration the solution by creating use of glass wool. Little drops of Molish, s reagent were added to filtration solution. Then violet colour appeared, this indicated to presence of Carbohydrates.
3 Quantative analysis: 2. 1 Alkaloid content: It is determined regarding to method described by 8. Evaporated combined extract on steam bath with space current to about 10 ml sum measured excess 0. 2N H2SO4 and continue evaporation to remove the solvent, cold and sum methyl. Then the excess acid H2SO4 was titrated with 0.
V1 X M1 = V2 X M2 two Mg or g = V2 X D. WT 3 103 X S Where: V1 = Volume of 0. 2N H2SO4, M1 = Concentration of H2SO4 0. 2N, V2 = Volume of NaOH, M2 = Concentration of NaOH 0. WT = Molecular mass of NaOH, S= Mass of sample and 103 = conversion factor from volume to weight.
2 Tannin content: It is determined regarding to method described by 11. 2 gram of extracted sample was placed in 50 ml of conical flask and 10 ml of 1% HCL was added. The contents of flask were shaken for 20 minutes, and then contents regarding the flask were centrifuged at 3000 rpm for six minutes. Then contents of flask were incubated at 30 oC for 20 minutes. The spectrophotometer was view at 500 nm and then scale view zero with the blank solution.
Finally, readings were taken 3 times and averaged. % = C X 100 two 10 X S Where: C. = Catechin equivalent, C = Concentration corresponding to optical density, 10 = Volume of extract ml and S = Mass of sample. 0 Conclusions and Discussion 3. 1 Qualitative analysis: The qualitative test of lime and black tea that extracted by water, ethanol and petroleum ether for the alkaloid, anthraquiaones, saponins, tannin, cardenolides, steroid, flavonoids, coumrine, and carbohydrates were shown in Table 1.
The findings revealed that the alkaloid, tannin, steroid, flavanoid in 3 extractions water, ethanol and petroleum ether of lime and black tea are available. Thecardenolide in 3 extractions ethanol and petroleum ether of lime and black tea are presence but thecardenolide in h2o extraction is not located in lime and black tea. These conclusions indicated thatcardenolide is not dissolved in h2o but dissolved in ethanol and petroleum ether. In addition to that the anthraquiaones and coumarin 3 extractions water, ethanol and petroleum ether of lime and black tea are not available. The saponin in lime and black tea extracted by h2o is present, but saponin in lime and black tea extracted by ethanol and petroleum ether is not present These findings revealed that lime and black tea not ever contain anthraquiaones and coumarin.
These conclusions are supported by those conclusions provided by 12. 2 Quantative analysis: Table 3 indicated that the alkaloid content in lime tea in water, ethanol and petroleum extracted is 5. While the alkaloid content in black tea in water, ethanol and petroleum extracted is 6. The conclusions revealed that alkaloid in most lime and black tea in h2o extracted is higher than in ethanol and petroleum ether extracted, but there is no significant difference in 3 solvents extracted for lime tea at P? 0. This indicated that extraction of alkaloid is not effected by categories of solvents.
In addition to that the conclusions indicated that tannin content in lime and black tea for h2o extracted is similar, tannin content in lime tea for ethanol extracted is higher than in black tea. The conclusions revealed there significant difference at P? 0. The variation in alkaloid and tannin content is attributed to kind of solvent, kind of tea and process of tea. Conclusion and Recommendation: We concluded that the lime and black tea contain alkaloid, tannin, steroid, flavonoid, cardenolides, saponins and carbohydrate. of alkaloid and tannin are varied regarding to kind of solvent used.
Our recommendation, the farther read in lime and black tea should be done particularly antimicrobial study. References two Berube P. Effects of encapsulated lime tea and Guarana extract containing a mix of epigallocateechin 3- gallate and caffeine on 24 hours life expenditure and fat oxidation in men. Double blinded, placebo- controlled trial of lime tea extracts within the clinical and histological appearance of photo aging skin. Effects of dosing condition on the oral bioavailability of lime tea catechins subsequent to single=dose administration of polyphenols in well individuals.
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Cholesterol lowering effect regarding the aflavins- Enriched tea extract: a randomized controlled trial. Association of Official Analytical Chemists. Official Methods of Analysis. Change in tannin in ripening fruit. Phyto chemistry 2, 371 10 FAO 1991.
Book to Specification: general notices, general analytical techniques, identification tests, test solution and other reference materials. FAO Food and Nutrition cardboard six Rev. A critical evaluation regarding the Vanillin Reactions as an assay for tannin in sorghum rain. Medicinal Plant Constituents. , General Organization for University and Schoolbooks.
Table 1? Chemical constituents that available + ve and non-available - ve in lime and black tea. Solvent extraction or Compounds H2o extraction Ethanol extraction Petroleum ether extraction Lime tea Black tea Lime tea Black tea Lime tea Black tea two or Alkaloid + ve + ve + ve + ve + ve + ve 3 or Anthraquinones - ve - ve - ve - ve - ve - ve 4 or Cardenolides - ve - ve + ve + ve + ve + ve 5 or Tannin + ve + ve + ve + ve + ve + ve six or Saponins + ve + ve - ve - ve - ve - ve seven or Coumarin - ve - ve - ve - ve - ve - ve 7 or Flavonoids + ve + ve + ve + ve + ve + ve 8 or Steroid + ve + ve + ve + ve + ve + ve 9 or Carbohydrates + ve + ve + ve + ve + ve + ve Table 2? Alkaloid and tannin content in extracted tea solvent water, ethanol and petroleum ether of lime and black type. Kind of solvent H2o extraction Ethanol extraction Petroleum ether extraction Kind of tea Lime tea Black tea Lime tea Black tea Lime tea Black tea Alkaloid Mg or g 5. 8 Each cost is average of 3 replicates expressed on hard mass basis.
