DETERMINATION AND VALIDATION METHOD OF TOTAL FLAVONOID CONTENT AND TOTAL PHENOLIC CONTENT OF ETHANOLIC EXTRACT OF MANGOSTEEN (Garcinia mangostana L.) LEAVES AS NATURAL PRESERVATIVES CANDIDATE BY USING SPECTROPHOTOMETRIC METHOD Diniatik, Indri Hapsari, Mil
On Education, Technology and Science (NETS) 2013 Purwokerto, December 28 th
, 2013
DETERMINATION AND VALIDATION METHOD OF TOTAL FLAVONOID
CONTENT AND TOTAL PHENOLIC CONTENT OF ETHANOLIC EXTRACT OF
MANGOSTEEN (*arcinia mangostana L.) LEAVES AS NATURAL
PRESERVATIVES CANDIDATE BY USING SPECTROPHOTOMETRIC METHOD
*Diniatik, Indri Hapsari, Milani Tiara, Ayu Meidyawati, Suhri Nurhidayat
*E,mail: diniatik@yahoo.com.au
Faculty of PharmacyFakultas Farmasi Muhammadiyah Univercity of Purwokerto
Jalan Raya Dukuhwakuh PO BOX 202, Purwokerto 53182, Jawa Tengah, Indonesia
Abstract Mangosteen has great potential as a natural preservative, it is necessary to ensure the content of compounds that have the potential levels. This study aimed to determine levels of total flavonoid content and total phenolic content and the validation methods. Determination and validation method used spectrophotometric methods, the methods of hydrolysis on total flavonoid content and the folin ciocalteu reagent method in the determination of phenolic total. Spectrophotometric method can be used for the determination of total flavonoid content with the results of the validation are eligible. The result of determination of total flavonoid content in 50% ethanol extract was 26.80 ± 15.9%. Spectrophotometric method can be used for the determination of total phenolic content with the results of the validation are eligible. The result of determination of total phenolic content in 50% ethanol extract was 71.94 ± 12.37%.
Keywords: ethanolic extract of Garcinia mangostana leaves, spectrophotometri, total flavonoid content, total phenolic content
INTRODUCTION
Currently the plant has many uses by humans, as building materials, food, medicine. The utilization of plants as additional material is limited, natural dyes, natural flavoring, natural smell giver, but as a natural preservative is limited utilization. Natural preservatives from plant could be developed further because of the danger of side effects and limitations of synthetic preservatives when used in large quantities is not good effect. The use of plants as a natural preservative has economic potential, as more people aware the dangers of synthetic preservatives so that people interested in eating organic food and organic processed food.
In processed foods, brown sugar, sodium metabisulfite used as a preservative, which has some potentially damaging effects of such toxic genes (mutagens) tehadap bacteria have a tendency toward carcinogens, long,term use produces damaging organs (skin allergy, liver, metabolism and urinary system, asthma), toxic to the respiratory tract, skin, eyes, reproductive organs. Toxicity in rat LD50 1131 mg / kg and guinea pig skin LD50> 1000mg/kg. This spurred the possibility that relatively safer preservatives, with long,term continuity of production.
Mangosteen has informed empirically used as a preservative, especially the skin of the fruit. Thus the excavation of the main part of the leaves of this plant which is part of the easy and cheap to produce natural preservative be more likely to meet the needs of sustainable natural preservative. In the research that has been done further examine the skin of the fruit, but the research that has been done by Diniatik and Supaman (2011) reported the antioxidant activity of the leaves better than the skin of the fruit. Continued research and Diniatik Supaman (2011), reported the antibacterial activity was also the leaves better than the skin of the fruit. The first phase of the study showed antioxidant activity, antifungal and antibacterial of 50% ethanol extract of mangosteen leaves have the best potential. So it is very interesting to do further research to the second and third stages. There are three problems of this resears:
1. How validation method and determination total flavonoid conent in the ethanol extract of mangosteen leaves?
2. How validation method and determination total phenol content in the ethanol extract of mangosteen leaves?
3. How determination methods xanthon content in ethanol extract of mangosteen leaves?
METHOD Variables in The Research
The definition of operational variables in this study include independent variables used were 50% On Education, Technology and Science 2013 Purwokerto, December 28 th
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3. Preparation of ethanol extract of leaves of *$ mangostana L.
5. Determination of total phenolic content and validation method
The workings of hydrolysis is as follows: 100 mg of extract was weighed precisely and put in a round bottom flask. Added hydrolysis system, ie 1.0 ml solution of 0.5% w/v heksanametilentetramin, 20.0 ml of acetone and 2.0 ml of HCl (25%) in water. Do hydrolysis by heating to boiling (used cooling water or reflux) for 30 minutes. Hydrolysis resulting mixture was filtered using a cotton swab into a 50.0 mL volumetric flask. Hydrolysis residue plus 10 ml of acetone to boil again briefly, do as much as 2 times and the filtrate collected all into the flask. After a cold flask, then the volume is turned up just 50.0 ml, beaten flat. Hydrolysis filtrate was extracted by 20 ml whipped with a separating funnel, first with 10 ml of ethylacetate. Then 2 times with 5 ml of ethyl acetate, and the ethyl acetate fraction was collected into a 25.0 mL volumetric flask, until it finally added 25.0 ml of ethyl acetate (POM, 2000). Validation of a method to determine the linearity, LOD (Limit of Detection) and LOQ (Limit of Quantification).
The principle of this method is that the levels are defined as flavonoid aglycone by hydrolysis method, spectrophotometric measurement at a wavelength of maximum, after reacted with AlCl3 to improve selectivity and added heksanametilentetramin.
4. Determination of total flavonoid content and validation method
Extracts of juice and ethanol were evaporated to a thick consistency is obtained which still can be cast and then be weighed. Preparation of ethanol extract of leaves of G. mangostana done at the Laboratory of Pharmaceutical Biology, Muhammadiyah University of Purwokerto.
Extracts of juice were made by using fresh leaves that have been washed and then 3 kg of leaves blended with 3 liters of distilled water until soft portion. Filtered with a flannel cloth.
Crude leaf weighed +3 kg powder made by grinding and sieving process. +750 grams of dry powder was weighed and then macerated. Maceration is done by immersing the crude drug powder in the solvent. In this research, to improve the effectiveness of the extraction is done stirring and remaceration, macerated for 2 x 24 hours with a comparison between the crude drug with 50% ethanol is 1:5 for the first day, and 1:4 for the second day. The way is as much as 750 grams of powder botanicals are macerated with ethanol 50% as much as 7.5 liters, then precipitated, poured and squeezed. Dregs obtained macerated again with as much as 3 liters of ethanol.
The equipments were used in this study: Maceration equipment, Bekker Glass (Pyrex) glass and other tools, Rotary Evaporator, electric cooker, separating funnel, UV spectrophotometer, micropipette, filter paper.
ethanol extract. Dependent variable is the level of flavonoids, phenols and total ethanol extract of leaves xanthon G. mangostana L. And the control variable is validated assay method with spectrophotometric method.
Materials used in this study: Ethanol, ethyl acetate, distilled water. acetone, methanol pa., several standards that rutin, gallic acid and alpha mangosteen.
Java. Drying is done by putting materials that have been washed clean in the winnowing, then covered with a black cloth that is not exposed to direct sunlight. After it dried under the sun to dry. The dried crude drug (taking into account the requirements of the maximum water content in crude drug) diserbuk and placed in a brown bottle dry.
2. Collection of materials Crude leaf G. mangostana Leaves of G. mangostana were taken Banyumas regency, Central
Mangosteen (G. mangostana) were used in the research determined in the Laboratory of Biology, Faculty of Education Studies Teacher Training and Education, University of Muhammadiyah Purwokerto. The results of determination convey that the plant specimen is Garcinia mangostana L. plant of the family Clusiaceae. The results of the determination by the book Flora of Java Vol II (Backer & Van Den Brink, 1965).
1. Plant Determination
The Way of The Research
The sample was used in this study is the ethanol extract of leaves of G. mangostana L. standard rutin, gallic acid and alpha mangosteen.
Sample and Standard
Total phenol content in the 50% ethanol extract of mangosteen leaves. Weighed 0.3 grams of extract were diluted to 10 ml denganmetanol: water (1:1). Pipetted 0.2 ml of extract solution and tambahkan15, 8 ml aquabidest add 1 ml of reagent Folin,Ciocalteu shake. Let stand for 8 minutes then add 3 ml of 20% Na2CO3 into the mix, let stand solution for 2 hours at room temperature. Measure with spektrofotometeriUV,Vis absorbance at 765 nm maximum absorption panjanggelombang which On Education, Technology and Science (NETS) 2013 Purwokerto, December 28 th
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will provide complex biru.Lakukan 3 times repetition so that diperolehhasilnya phenol obtained as mg gallic acid equivalent/g fresh sample (Scrambled, 2006). Validation of a method to determine the linearity, LOD and LOQ.
G . Analysis
Datas were obtained from measurements using a standard curve of linear regression equation on the relationship curve between the absorbance of the extract concentration (Gulluce, et al., 2006).
RESULT AND DISCUSSION Plant Determination
Mangosteen (G. mangostana) were used in the study determined in the Laboratory of Pharmaceutical Sciences Faculty of Pharmacy, Muhammadiyah University of Purwokerto. The purpose of determination to get the truth with a clear identity of the studied plants and avoid mistakes in the collection of primary research materials. The results of determination stating that the plant specimen is really,really plant Garcinia mangostana L. of the family Clusiaceae. The results of the determination by the book Flora of Java Vol
II (Backer and Van Den Brink, 1965).
Materials Collection and Crude Preparation of Leaves mangosteen (*$ mangostana)
In this study the use of mangosteen (Figure 1) were taken at the District Somagede, Banyumas. Fresh leaves of mangosteen taken in May in the morning. The leaves are plucked fresh mangosteen leaves was being taken from the 30,year,old trees since planting, the leaves taken after post,harvest fruit. The leaves are taken is elderly, because it is expected that the chemical content obtained were optimal (Anonymous, 1985). Decision were done at a certain time and place to avoid wide,range of chemical constituents due to differences in environmental conditions, objec soil, and climate.
Figure 1. Mangosteen plant (G mangostana) Fresh mangosteen leaves were obtained, washed with running water to remove any dirt or contaminants in the form of soil, or other materials on the the leaves. Selected the leaves (15 kg) is good for the next aerated. Partially dried mangosteen the leaves in the sun covered with black cloth, for the purpose of extraction with ethanol. During heating, the material is not accumulated and inverted laid,back that evenly heating and drying process takes place quickly. The dried mangosteen leaves on the seventh day. Leaves the rest was made in the form of mangosteen juice (Anonymous, 1985). Drying is done until the moisture content is less than 10% or until the leaves are easy to be destroyed when squeezed. The purpose of drying is to prevent the growth of fungi or microorganisms and the decomposition of active compounds by enzymatic reactions and hydrolysis process because of high water content, so that the resulting crude drug is not easily damaged so it can be stored in a relatively long time. Dried Simplicia were grinded then by using a blender to minimize the surface area that contacts the surface of the particles of crude drugs with greater solvent and dissolving more optimal.
Crude further were sieved using a sieve mesh 20/40 which means as much as 100% dry simplicia passes on 20 mesh sieve, then as much as 40% from 100% dry simplicia 40 mesh sieve, so that 500 grams of dried botanicals mangosteen the leaves and bark as much as 300 grams 20 mesh sieve and sieve as much as 40 grams of 200. In general screening process is important in the extraction process, due to the presence of particle size reduction will expand the contact surface with the powder so penyari maximum and extraction becomes more active substances may be taken optimally.
Making Ethanol Extract of Mangosteen Leaf
Dissolution method used is maceration. This method is the simplest method because it is easy to do, inexpensive, does not require sophisticated equipment. Maceration is done by soaking the powder in a liquid botanicals penyari. In this research, to improve the effectiveness of the extraction is done stirring and remaserasi, macerated for 2 x 24 hours with a comparison between the crude drug with 50% ethanol is 1:5 for the first day, and 1:4 for the second day. The trick is as much as 750 grams of powder botanicals are macerated with ethanol 50% as much as 3.75 liters, then dienap,pour and squeezed. Dregs obtained again macerated with ethanol 50% as much as 3 liters.
In this study, the solven were used 50% ethanol. Solven 50% ethanol can be interesting compound , a relatively polar compounds such as
3
(figure 2).
, 2013
phenolic compounds, flavonoids, saponins, and other polar compounds contained in mangosteen the leaves, whereas ethanol is ethanol used in the market is technically possible it is recovered when evaporated with a rotary evaporator. Ethanol is more selective, non,toxic, neutral, good absorption, can prevent the growth of mold and bacteria, the heat required for concentration less. Juice obtained is evaporated over a water bath until a thick consistency. Evaporation is done to eliminate solven solution so as not to affect the next examination as shown in Table 1 below.
Tabel 1. The yield of ethanol extract Type
The weight of wet simplicia The weight of dry simplicia
Extract weight Rendemen ethanol extract 50% 3 kg 750 g 175,41 g 5,647%
Method Validation and Determination of Total Flavonoids Content
Total flavonoid content of ethanolic leaf extract of mangosteen determined using spectrophotometric methods that have been recommended in the book POM body Sandar Extract Parameters (Anonymous, 2000). The principle of this method is that the levels are defined as flavonoid aglycone by first hydrolyzed and subsequent spectrophotometric measurements by reacting AlCl
On Education, Technology and Science 2013 Purwokerto, December 28 th
occurs that causes a wavelength shift to the right, making it more selective. Hydrolysis (reflux) done to change into aglycone flavonoid glycosides flavonoids and glikon. This is done because the flavonoids in ethanolic herbal extracts contained in china patikan glycoside and aglycone form. Determination of total flavonoid levels aims to establish the levels of all flavonoids (total), with hydrolyzed, all will flavonoid aglycone flavonoids tertetapkan as advantageous because it derived flavonoid levels is certainly more appropriate. Wavelength shift toward longer bathochromic well as OH groups that had been closed off so that the sugar will slide more reactants to form bonds, the maximum wavelength becomes larger, making it more selective. The disadvantage is the long working procedures so that the possibility of error for greater progress, for it is the determination of the total flavonoid content patikan chinese herbs to prior validation of methods including linearity, precision, accuracy, detection limit of the test.
The standard used was rutin, quercetin and ramnoglukosa hydrolyzed form. After the quercetin was added AlCl
3 reagent to form a chelate complex that become has a larger maximum wavelength
- Ramnoglukosa Kuersetin AlCl
Rutin Kuersetin
3 Komplek Kuersetin dengan Al
Figure 2. Hydrolysis rutin into quercetin and reaction with AlCl
3 On Education, Technology and Science (NETS) 2013 Purwokerto, December 28 th
, 2013 . Preparation of Standard Curve and Linearity
Standard curve was obtained by varying the levels that have been hydrolyzed rutin (same method with the sample) was measured at a wavelength of 417 nm maximum. From the data obtained by spectrophotometric absorbance for each concentration (Table 2). The absorbance is plotted with each concentration to make the linear regression equation y = bx + equation a. From the relationship curve between absorbance and concentration will be obtained correlation coefficient (r) as a parameter to determine linearity. The results obtained equation has good linearity, because r count (0.9984) is greater than r table (0.811, for N = 6, 95% confidence level) (Hadi, 1996). Standard curve was obtained Y = 0.00491 x , 0.00726 is used to determine the total flavonoids content, linear regression curve can be seen in Figure 3.
Table 2. Absorbance and standard curve
No. Rutin content(mg/L) Absorbance (A) The result of linear regression
1.40 0,1938 r = 0,9984 a = ,0,00726 b = 0,00491 2. 80 0,3878
3. 120 0,5515 4. 160 0,8114 5. 200 0,9790 6. 240 1,1642
Figure 3. Standard linear regression curve of rutin that has been hydrolyzed
2. Determination of Limit of Detection
To determine the total flavonoid content in the sample was able to be detected by this method was measured the levels of several standard rutins that have been hydrolyzed. The measurement results can be seen in table 3. Limit concentration of standard solutions that can still be detected by spectrophotometric method is 1.25 mg/L. This means that the total flavonoid assay with a standard rutin which is hydrolyzed to the sample by spectrophotometric methods measure should be greater than 1.25 mg/L. Figure 4 shows the linear regression curves were created from data from the determination of the minimum limit of detection with a correlation coefficient of r = 0.9979.
Table 3. Absorbance data and results of the minimum limit of detection
No. Rutin content (mg/L) Absorbance Results of the minimum limit of detection
1.20 0,0933 r = 0,9979 a = ,0,001939 b = 0,004736 2. 10 0,0436 3. 5 0,0249
4. 2,5 0,0065 5. 1,25 0,0027 6. 0,06 ,0,0011
On Education, Technology and Science 2013 Purwokerto, December 28 th
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Figure 4. Linear regression curve assay
3. Determination of Precision Precision test is done by making six replication of a standard rutin that has been hydrolyzed. The absorbance obtained standard curve were plotted on the Y = 0.00491 x , 0.00726 to obtain calculated content, and then calculated the average concentration, standard deviation and coefficient of variation (Table 4). The coefficient of variation (CV) was 1.80%, less than 5% indicates that this method has a good precision (Peter & Richard, 2000).
Table 4. Absorbance data and the results of precision test
No. Absorbance Rutin Content (mg/L) The results of precision test
1. 0,5619 115,91 Rata,rata = 113,87SD = 2,05 CV = 1,80%
2. 0,5508 113,65 3. 0,5482 113,12 4. 0,5649 116,52 5. 0,5374 110,92 6. 0,5482 113,12
4. Accuracy Test of accuracy (closeness) is done by calculating the recovery rutin standard that has been hydrolyzed. Recovery percentage was calculated by making a comparison between content from the results of analysis with actual content. The absorbance were obtained, were plotted on standard curve to obtain calculated content, and then calculated the average content, standard deviation and recovery (Table 5). Value recovery (recovery) between 92.43% to 97.10%. These results meet the requirements due to the analyte content 0.01 <x <0.1% in the amount of 90,107%. The coefficient of variation are also eligible ie 1.80% (less than 5.3%) (Yowono & Indrayanto, 2005). This means the method meet the requirements, because especially the precision and accuracy have met the requirements, systematic errors which arise in the analysis can be avoided.
Table 5. The results of accuracy test
No. Actual rutin content (mg/L) Rutin content From the result of analysis (mg/L) The results of accuracy test (%) 1. 120 115,91 96,50 2. 120 113,65 94,70 3. 120 113,12 94,26 4. 120 116,52 97,10 5. 120 110,92 92,43 6. 120 113,12 94,26
5. Determination of Total Flavonoids Content
The results of the determination of the total flavonoid content 50% ethanolic extract of mangosteen leaves are calculated as hydrolyzed rutin can be seen in Table 6. The results of the assay 50% ethanolic extract of mangosteen leaves are 42.33%, 27.53% and 10.55% (w/w). On Education, Technology and Science (NETS) 2013 Purwokerto, December 28 th
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Table 6. The results of the assay was calculated as the total flavonoids rutin hydrolyzed
The origin mangosteen leaf area Absorbance Total flavonoid content mg/L Total flavonoid content%(b/b) In the extract The average of total flavonoid content%(b/b) In the extract
Banyumas 0,3251 67,69 42,33 26,80 ± 15,9 0,4261 88,26 27,53 0,1587 33,80 10,55
Mangosteen leaves are extracted with 50% ethanol derived from Banyumas region have relatively high levels of flavonoids, namely 26.80%.
Validation Method and Determination of Total Phenol Content
Total phenolic content of ethanolic leaf extract of mangosteen determined using spectrophotometric method referring to previous studies (Scrambled, 2006). Determination of total phenol content which aims to define all content phenols (total) in the ethanolic leaf extract of mangosteen. Prior to the determination of total phenol content to prior validation of methods including linearity, precision, accuracy, of detection limit of the test. Appellant used are Gallic Acid.
1. Preparation of Standard Curve and Linearity Standard curve was obtained by varying the content that have been hydrolyzed rutin (same method with the sample) was measured at a wavelength of 601 nm maximum. From the data obtained by spectrophotometric absorbance for each concentration (Table 7). The absorbance were plotted with each concentration to make the linear regression equation y = bx + equation a. From the relationship curve between absorbance and concentration will be obtained correlation coefficient (r) as a parameter to determine linearity. The results obtained equation has good linearity, because r count (0.9938) is greater than r table (0.811, for N = 6, 95% confidence level) (Hadi, 1996). Standard curve obtained is Y = 0.005007 x , 0.304 is used to determine content total phenols, linear regression curve can be seen in figure 5.
Table 7. Absorbance data and standard curve
No. Gallic acid content(mg/L) Absorbance (A) The result of linear regression
1. 52,86 0,5671 r = 0,9938a = 0,304 b = 0,005007
2. 78,94 0,6876 3. 104,80 0,8558 4. 130,43 0,9314 5. 155,84 1,1237 6. 181,03 1,1886
Figure 5. Linear regression standard curve of gallic acid standard
2. Determination of Limit of Detection
To determine the total phenol content in the sample was able to be detected by this method of was measured multiple content gallic acid standard. The measurement results can be seen in the following table. Limit content of standard solutions that can still be detected by spectrophotometric method is 7,723 mg / L. This means that the assay of phenol with standard gallic acid hydrolyzed in samples with spectrophotometric methods measure should be greater than 7,723 mg / L. Figure 6 shows the linear regression curves were created from data from the determination of the minimum limit of
The results of accuracy test (%) 1. 155,84 154,46 99,114 2. 155,84 160,23 102,81 3. 155,84 158,47 101,68 4. 155,84 160,09 102,73 5. 155,84 162,71 101,408
No. Absorbance Gallic acid Content (mg/L) The results of precision test 1. 1,0774 154,46 Rata,rata = 159,192
No. Actual gallic acid content (mg/L)
Gallic acid content
From the result of analysis (mg/L)
Table 10. The results of accuracy test
Value of recovery (recovery) between 99.114% to 102.81%. These results meet the requirements due to the analyte content 0.01 <x <0.1% in the amount of 90,107%. The coefficient of variation are also eligible ie 1.91% (less than 5.3%) (Yowono & Indrayanto, 2005). This means the method meet the requirements, because especially the precision and accuracy have met the requirements, systematic errors which arise in the analysis can be avoided.
4. Determination of Accuracy Accuracy test (closeness) is done by calculating the recovery (recovery) rutin standard that has been hydrolyzed. Recovery percentage was calculated by making a comparison between content results of analysis with actual content. The absorbance obtained were plotted on standard curve to obtain calculated content, and then calculated the average content, standard deviation and recovery (Table 10).
2. 1,1063 160,23 3. 1,0975 158,47 4. 1,1056 160,09 5. 1,1187 162,71 6. 1,2373 186,39*
SD = 3,048 CV = 1,91%
Table 9. Absorbance data and ther result of precision test
On Education, Technology and Science 2013 Purwokerto, December 28 th
3. Determination of Precision Precision test is done by making six replication of standard gallic acid which has been hydrolyzed. The absorbance obtained standard curve were plotted on the Y = 0.005007 x , 0.304 to get calculated content, and then calculated the average content, standard deviation and coefficient of variation (Table 9). The coefficient of variation (CV) was 1.91%, less than 5% indicates that this method has a good precision (Peter & Richard, 2000).
Figure 6. Linear regression curve of limit of detection
2. 12,357 0,281 3. 9,268 0,132 4. 7,723 0,077 5. 6,179 ,0,0044
a = ,0,2311 b = 0,0391
No. Gallic acid content (mg/L) Absorbance Results of the minimum limit of detection
1. 15,446 0,353 r = 0,99132Table 8. Absorbance data and the result of limit of detection
, 2013 detection with a correlation coefficient of r = 0.99132.
- Tidak dimasukkan karena dianggap alien
Purwokerto, December 28 , 2013
5. Determination of Total Phenol Content be seen in the following table. The results of the
The results of the determination of total assay 50% ethanolic extract of mangosteen leaves phenolic content 50% ethanolic extract of are 59.23%, 83.96% and 72.64% (w / w). mangosteen leaves are calculated as gallic acid can
Table 11. The results of the determination of total phenol content was calculated as gallic acid
The origin Absorbance Total gallic acid Total gallic acid The average of total
mangosteen content content%(b/b) gallic acidleaf area mg/L In the extract content%(b/b)
In the extractBanyumas 04622 31,59 59,23 71,94 ± 12,37 0,5287 44,87 83,96 0,4988 38,90 72,64
Mangosteen leaves are extracted with 50% ethanol spectrophotometric method is by using a standard derived from Banyumas area has a large content of curve alpha mangostin. With the data y = 0.019 x phenol is 71.94%. +0.057 to 0.999 linearity. The results of the assay xanthon total 50% ethanolic extract of mangosteen
Determination of Total Xanthon leaves was 1.56%, 2.41% and 1.59% (w / w).
Xanthon total concentration on ethanolic leaf extract of mangosteen determined by using the Table 12. The results of the determination of the total xanthon calculated as alpha manggosteen
The origin Absorbance Total xanthon Total xanthon The average of total
mangosteen content content%(b/b) xanthon content%(b/b)
leaf area mg/L In the extract In the extractBanyumas 0,3691 16,42 1,56 5,56 ± 0,48 0,5656 26,77 2,41 0,3739 16,68 1,59
Mangosteen leaves are extracted with 50% 2006c; nonymous, 2002b). Rainfall in Banyumas ethanol derived from Banyumas has a total xanthon district that is located on the slopes of Mount content relative to the average alpha mangostin is Slamet is quite high, 3,439 mm every year, making 5.56 ± 0.48% w / w%. Spectrophotometric assay the soil fertile, mangosteen leaves in this area the method is the same as Aisha et al study (2013) width, height and fertile. which establishes the fruit skin extracts, and high performance liquid chromatography method on CONCLUSIONS AND RECOMMENDATIONS
1. Conclusions fruit skin extracts on research Yodhnu et al (2009).
Levels of total flavonoids (26.80 ± 15.9% w / The results of this study it can be w), total phenols (71.94 ± 12.37% w / w) and total concluded: xanthon (5.56 ± 0.48% w / w) high in Banyumas
a. Spectrophotometric method can be used for because lower temperatures (air temperature ranged determination of total flavonoid content with from 21.4 to 30.9°C with an average temperature of the results of the validation are eligible. The 26.3°C) and higher light intensity. Mangosteen results of the determination of the total leaves taken from Somagede located in the flavonoid content in the 50% ethanol extract mountains near Slamet. Banyumas is located at an was 26.80 ± 15.9% altitude of 25 to 500 meters above sea level, there b. Spectrophotometric method can be used for
Slamet mountain with a summit elevation of sea determination of total phenolic content with level about 3,400 feet and is still active. Weather the results of the validation are eligible. The and climate in Banyumas as classified in the results of the determination of total phenolic equatorial southern hemisphere still has a wet content in 50% ethanol extract was 71.94 ± tropical climate, situated between the slopes of the 12.37% mountains away from the surface of the ocean shore c. Spectrophotometric method can be used to or sea wind effect is not so apparent (nonymous, assay total xanthon with linearity results
On Education, Technology and Science 2013 th
Purwokerto, December 28 , 2013 qualify. Xanthon total assay results in 50% Backer, C. A, dan Van den Brink, Jr. R. C. B. 1968.
ethanol extract was 5.56 ± 0.48% Flora of Java, Vol. II, Published Under The Auspeces of The Rijkher Barium. Lieden.
2. Recommendations
Bo, T. & Liu, H., 2004, Separation methods for This research is still a weakness, it is pharmacologically active xanthones, Journal necessary to study more about: of Chromatography B, Volume 812, Issues 1– determination of the best formulations to be used 2, 5 December 2004, Pages 165–174. preservatives disa society. Diniatik dan Suparman, 2010, Uji Antioksidan
Ekstrak Daun dan Kulit Batang Manggis
ACKNOWLEDGEMENTS (Garcinia mangostana L) dengan Metode
This research was conducted as a research DPPH, Laporan Penelitian LPPM UMP.grant from the Private Higher Education Hadi, S., 1996, Statistik 2, Jilid II, Cetakan XVI, Coordinating Region VI, the Ministry of Education Penerbit Andi Offset, Yogyakarta. and Culture, therefore, we say thank you profusely. Orak, H.H. 2006, Total Antioxidant Activities,
Phenolics, Anthocyanins, Polyphenoloxidase
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