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The effect of solvents and extraction time on total xanthone and antioxidant yields of mangosteen peel (Garcinia mangostana L.) extract
Article in Drug Invention Today · January 2018
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The effect of solvents and extraction time on total xanthone and antioxidant yields of mangosteen peel (Garcinia mangostana L.) extract
Andri Kusmayadi1,2*, Lovita Adriani1, Abun Abun1, Muchtaridi Muchtaridi3, Ujang Hidayat Tanuwiria1
INTRODUCTION
Mangosteen (Garcinia mangostana Linn.) is a premium commodity of Indonesian local fruit with import potential. The biggest mangosteen producer in Indonesia is West Java Province with approximately 69.314 of 203.103-ton national harvest in 2015. The yield from other regions included 29.748 ton in East Java, 20.339 ton in West Sumatra, 12.190 ton in Central Java, 10.660 ton in Bali, 9.760 ton in Banten, 7.947 ton in North Sumatra, 7.078 ton in West Nusa Tenggara, 5.419 ton in Lampung, and 5.290 ton in South Sulawesi.[1] West Java is, therefore, contributed one-third of national demand of mangosteen (>33%).
In West Java, Tasikmalaya Regency is the biggest mangosteen producer in 2.561 Ha productive land.
During mangosteen season, the harvest is plenty and the peels are potential waste because 65% of mangosteen
Research Article
1Department of Animal Science, Faculty of Animal Science, Universitas Padjadjaran, Sumedang-45363, Indonesia,
2Department of Animal Science, Faculty of Agriculture, Universitas Perjuangan, Tasikmalaya-46115, Indonesia, 3Department of Pharmaceutical Analysis and Medicinal Chemistry, Faculty of Pharmacy, Universitas Padjadjaran, Sumedang-45363, Indonesia
*Corresponding author: Andri Kusmayadi, Faculty of Animal Science, Universitas Padjadjaran, Bandung – Sumedang KM. 21. Jatinangor, Sumedang, Indonesia. Tel. +62-8977719392. E-mail: [email protected]
Received on: 18-05-2018; Revised on: 16-06-2018; Accepted on: 25-07-2018 Access this article online
Website: jprsolutions.info ISSN: 0975-7619
fruit is the peel. A fresh mangosteen consists of 17%
outer skin, 48% inner skin, 31% flesh, and 4% stem and petals.[2] Besides high yield percentage, mangosteen peels also contain secondary metabolite or xanthone which is higher than the other parts of mangosteen.
Xanthone is a polyphenol compound with a chemical structure containing aromatic tricycle.[3]
Mishima et al.[4] reported that most xanthone in mangosteen peel is α-mangostin. Moreover, phenolic is more abundant in mangosteen peel than the aril.[5]
Palakawong et al.[6] suggested that mangosteen peel contains the most xanthone - the secondary metabolic compound, with multifunctions such as antioxidant.
Besides acting as an antioxidant, xanthones in mangosteen also play a role as pharmacokinetic agent, anti-obesity, anti-parasite, anti-inflammation, anti- allergy,[7] anticancer,[8,9] and also antibacteria.[10]
Xanthone can be isolated to obtain the benefit through the accurate method of extraction.[11]
Extraction can separate the important components such as antioxidant compound from the parts of the plant.[12] Selecting the appropriate method of ABSTRACT
This study aims to evaluate the effect of different solvents and extraction time on total xanthone and antioxidant yield of mangosteen peel extract. The research was conducted by extracting mangosteen peel using seven solvents: Ethanol, acetone, ethyl acetate, methanol, hexane, acetic acid, and aquadest at different times: 24, 36, and 48 h. Total xanthone was examined using visible UV spectrophotometer at the particular wavelength, and antioxidant yield was observed from the extract and then dried using rotary evaporator and freeze dryer. The result reported that the solvents and extraction time significantly (P < 0.01) affected total xanthone and antioxidant yield. Mangosteen peel extracted by acetone shows the best results on total xanthone at 48 h. Meanwhile, mangosteen peel extracted by ethanol for 24 h was the best result on antioxidant yield. Total xanthone levels and antioxidant yield are affected by solvents and extraction time. Total xanthone levels were best found at long extraction times, while the best levels of antioxidant yield were found at short extraction times. Acetone and ethanol solvents which have medium polarity index (5.1 and 5.2, respectively) show the highest levels of total xanthone and best antioxidant yield compared to other treatments.
KEY WORDS: Antioxidant yield, Mangosteen peel extract, Solvent, Extraction Time, Total Xanthone
Andri Kusmayadi, et al.
extraction must consider the characteristics of the compound to be extracted. Mangosteen peel, as well as the other natural resource, is characterized by (a) heterogenous shape, (b) abnormal frequency of distribution of the properties, (c) various composition based on the species, growth condition, and level of ripeness, and (d) vulnerability to chemical change, humidity, respiration, and enzymatic activity. There are several contributing factors to the effectiveness and efficiency of extraction process such as the types of solvent and polarity index,[11] extraction time, solvent concentration and ratio, temperature, and the particle size of the solid matrix.[13] However, the most influential factor in the amount of extractable bioactive is the type of solvent and extraction time.[14]
Selecting the type of solvent in the extraction process must consider several factors such as availability, low price, physically and chemically stable, neutral reaction, and impotent to bioactive compound.
Extraction time needs to be calculated so that the bioactive compound can be isolated optimally using the most efficient solvent. Extraction was conducted in a single or mixed solvent at particular time (1–2 days in general) without heating.[15] The combined design of solvents and extraction time can impact the antioxidant activity of mangosteen peel extract. To obtain the optimum antioxidant activity in mangosteen peel extract, a sound preparation for the condition and method of extraction must be well considered.[3]
MATERIALS AND METHODS
Materials
Mangosteen peel was obtained from Puspahiang, Tasikmalaya, West Java, Indonesia. The standard of α-mangostin and xanthone was purchased from Sigma.
Solvents used ethanol 96%, acetone, ethyl acetate, methanol, hexane, acetic acid, and aquadest.
Instrumentation
Magnetic stirrer (IKA® C-MAG HS7), UV-vis helios-α spectrophotometer, spray dryer, hair dryer, freeze dryer, vortex, analytical scales, oven, Erlenmeyer, rotary evaporator, homogenizer, miller, pH meter, filter paper Whatman No. 41, and glassware were used.
Procedure
Total Xanthone Test.
Creating Standard Α-Mangostin Solution
10 mg of standard α-mangostin solution was diluted with methanol in a 100-ml volumetric flask to reach the level mark to obtain 100 μg/ml concentration.
100 μg/ml from base solution was pipetted 0.2; 0.4; 0.6;
0.8 and 1 μg/ml respectively into the 10 ml volumetric
flask then diluted with methanol to reach concentration 2, 4, 6, 8 and 10 μg/ml, respectively, and measured for the absorbability using UV spectrophotometer at maximum absorbability wavelength 243 nm.[16]
Each extraction from the extracted mangosteen peel was weighed 50 mg and put in the volumetric flask, added with methanol to reach 50 ml level mark to obtain 1 mg/ml concentration (1000 μg/
ml), and then was filtered using Whatman filter paper No. 1. 1 ml of the concentrate was put in the volumetric flask and added with methanol up to the level mark to obtain a 10 μg/ml concentration. The absorbability was measured using UV spectrometry at the maximum absorbability wavelength 243 nm.
Total xanthone was calculated using the formula as follows:
c= A µ×b
Total xanthone mg/ml sample
( )
=cxBMX xFPml sample 10
A = Absorbability b = Cuvette width (mm)
ε = Emissivity of α-mangostin (3.16 × 103 L/mol) c = Extract concentration (mol/L)
BM = Relative mass molecule α-mangostin (410.47 g/mol)
FP = Solvent fraction.
Antioxidant Yield
Yield value was measured post-extraction at 24, 36, and 48 h. The calculation formula of the antioxidant yield was as follows: [17]
Yield of water Extracted water g Initial amount of water
=
( )
iin samples g
( )
(1)
Yield of extract Extracts g Dry samples g
=
( )
( )
(2)Statistical Analysis
Total xanthone and antioxidant yield from each treatment was tabulated and analyzed through t-test and ANOVA with SPSS software series 25.0 and in case of significant difference continued with Duncan’s multiple range test.
RESULTS AND DISCUSSION
Total Xanthone
Based on the statistical analysis in Table 1, the types of solvent and extraction time had a highly significant
impact (P < 0.01) on total xanthone of mangosteen peel extract. The types of solvent had the ability to isolate different xanthones in mangosteen peel extract due to the different characteristics. The longer the extraction occurred, the higher the total xanthone.
The result of total xanthone analysis suggested that mangosteen peel extracted using acetone solvent for 48 h had the highest xanthone (32.825 ± 1.919) while water solvent produced the lowest xanthone (19.460 ± 0.666). The result was in line with the study of Chhouk et al.[18] that xanthone was naturally insoluble in water and therefore difficult to be extracted using water.
Xanthone was, in fact, soluble in organic solvent with a moderate polarity such as acetone, ethanol, methanol, and ethyl acetate. It was evidenced from the present study that ethanol and methanol solvents produced the similar total xanthone with acetone, i.e., 30.669 ± 0.151 and 31.706 ± 0.667, respectively.
Total xanthone in the present study was lower than that of Aisha et al.,[16] in which methanol, ethyl acetate, and ethanol had 59.9 ± 0.1%, 79.1 ± 0.4%, and 86.5 ± 0.5%, respectively. It was because extraction method significantly influenced the extracted total xanthone.[19]
The absence of xanthone purification (α-mangostin) contributed to the accumulated absorption at the different wavelengths between components. It is because the dominant xanthone compound in mangosteen peel is α-mangostin, over a half of total xanthone.[20] Accordingly, the analysis of total xanthone using the approach of α-mangostin compound and high-performance liquid chromatography method produced a more accurate result.[21]
Table 1: Mean value of total xanthone of mangosteen peel extracted using different solvents and extraction time
Treatment Mean of total
xanthone
EtOH, 24 h 29.328±0.573b
EtOH, 36 h 30.116±1.177cde
EtOH, 48 h 30.669±0.151cdef
Ace, 24 h 29.158±1.243a
Ace, 36 h 31.025±1.100a
Ace, 48 h 32.825±1.919a
EtOAc, 24 h 27.262±0.173def
EtOAc, 36 h 27.265±0.038cde
EtOAc, 48 h 27.377±0.038ef
MeOH, 24 h 29.441±1.498bc
MeOH, 36 h 29.506±2.667bcd
MeOH, 48 h 31.706±0.667cde
HX, 24 h 20.173±0.713f
HX, 36 h 20.879±1.897ef
HX, 48 h 21.445±1.757ef
AcetAc, 24 h 23.863±0.013cde
AcetAc, 36 h 23.973±0.054cdef
AcetAc, 48 h 23.996±0.047cdef
Aqua, 24 h 19.460±0.666g
Aqua, 36 h 20.005±0.392g
Aqua, 48 h 20.646±0.626g
Values bearing different superscripts within row show a highly significant difference (P<0.01)
The applied extraction method was a simple method to extract the bioactive compounds in herbals.[19] Besides extraction, various alternative methods were found more effective to isolate the bioactive compound from plant extract. Extraction using Soxhlet, supercritical fluid extraction, subcritical water extraction, and ultrasonic extraction could produce a higher xanthone.
Subcritical ethanol extraction yielded more xanthone than the conventional method such as extraction and Soxhlet extraction.[11] The increase of xanthone level was linear with solvent concentrate and extraction time. The longer the time and temperature of extraction, the higher is xanthone solubility because the bioactive compounds were more soluble at higher than lower concentration due to the increasing area of extraction.[22]
Xanthone in mangosteen peel extract was essential for health[23] because xanthone is the source of antioxidant,[24] prevents diseases, and plays a role in alternative medication[10] such as for leukemia.[25]
Xanthone in mangosteen peel was easily damaged;
therefore, it should be protected using encapsulation made of purple yam starch in form of lozenges.[26]
Antioxidant Yield
Antioxidant yield of mangosteen peel extracted using different solvents at various times is presented in Table 2.
Table 2 shows that the types of solvent and time of extraction had a highly significant impact (P < 0.01) on the antioxidant yield of mangosteen peel extract. Different solvents had particular characteristics that produced different antioxidant yield - the higher the antioxidant
Table 2: Mean value of antioxidant yield of
mangosteen peel extracted using different solvents at a various times
Treatment Mean of antioxidant yield
EtOH, 24 h 11.74±0.496l
EtOH, 36 h 11.27±0.274jkl
EtOH, 48 h 11.03±0.417ijk
Ace, 24 h 10.68±0.517hij
Ace, 36 h 10.34±0.287h
Ace, 48 h 10.15±0.360gh
EtOAc, 24 h 9.67±0.496fg
EtOAc, 36 h 9.34±0.273f
EtOAc, 48 h 9.35±0.417f
MeOH, 24 h 11.41±0.517kl
MeOH, 36 h 10.53±0.287hi
MeOH, 48 h 10.43±0.360hi
HX, 24 h 9.46±0.496f
HX, 36 h 9.05±0.274def
HX, 48 h 8.70±0.417de
AcetAc, 24 h 9.25±0.518ef
AcetAc, 36 h 8.54±0.287d
AcetAc, 48 h 7.95±0.360c
Aqua, 24 h 6.84±0.496b
Aqua, 36 h 5.85±0.274a
Aqua, 48 h 5.80±0.418a
Values bearing different superscripts within row show a highly significant difference (P<0.01)
Andri Kusmayadi, et al.
yield, the more efficient the solvent. The ratio of solvent and herbals in the study was 10:1; therefore, the higher the extract antioxidant yield, the more effective and efficient the solvent to produce mangosteen peel extract.
The result indicated that 24 h extraction using ethanol resulted in the highest antioxidant yield.
The result of the present study supported the previous study that the yield of extraction was influenced by the solvent polarity.[27] The polarity index of the solvents in the study was 5.2% in ethanol, 5.1% in acetone, 4.4%
in ethyl acetate, 5.1% in methanol, 0.0% in hexane, 6.2% in acetic acid, and 9.0% in aquadest.[28] Acetic acid and aquadest had a high polarity index but yielded the lowest antioxidant due to the low solubility.[18] In contrast, ethanol, acetone, and methanol had similar polarity index (5.2–5.1) and high yield. It was evidenced that the moderate polarity was more efficient to produce the powder extract of mangosteen peel.
Ethyl acetate had a lower polarity index (4.4) and lower yield than that of ethanol, acetone, and methanol. It indicated that ethyl acetate was inferior to produce mangosteen peel extract due to lower polarity. Similarly, hexane with 0.0 polarity index had a considerably low yield due to the non-polar characteristic. The polarity index of solvent, therefore, directly affects the extraction process because it improved the solubility of antioxidant compound that eventually affected the antioxidant yield from extraction.[29]
Furthermore, the time of extraction affected the yield of mangosteen peel extract. Table 2 shows that the longer the time, the lower the yield. Ballesteros et al.[13]
reported that the contributing factors to the efficiency of extraction were: the types and concentration of solvent, ratio solvent:solids, temperature, particle size of the solid matrix and maceration time. The longer the solvent was in contact with mangosteen peel, the lower the antioxidant yield because the longer contact would accumulate the sediment and decrease the filtrate.
The result suggested a higher yield than that of Manimekalai et al.[30] who reported that the value of yield of purple mangosteen peel extracted by various types of solvent has a very low value. Methanol solvent was the optimum solvent with yield extract of the outer part of mangosteen peel (3.5%), followed by chloroform (1.5%), hexane (1%), and ethyl acetate (1%). The inner part of mangosteen peel produced the maximum yield from extraction using methanol (1.5%), followed by ethyl acetate (1.5%), chloroform (1%), and hexane (1%).
CONCLUSION
The solvent and extraction time resulted in a highly significant effect (P < 0.01) on total xanthone and antioxidant yield. Mangosteen peel extracted by acetone at 48 h shows the best results on total xanthone.
Meanwhile, mangosteen peel extracted by ethanol for 24 h gives the best result on antioxidant yield. Total xanthone levels and antioxidant yield are affected by solvents and extraction time. Total xanthone levels were best found at long extraction times, while the best levels of antioxidant yield were found at short extraction times. Acetone and ethanol solvents which have medium polarity index (5.1 and 5.2, respectively) show the highest levels of total xanthone and best antioxidant yield compared to other treatments.
ACKNOWLEDGMENT
The authors express their gratitude to Lembaga Pengelola Dana Pendidikan, Ministry of Finance, Republic of Indonesia, for the bestowed grant for the research.
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Source of support: Nil; Conflict of interest: None Declared
