Research article
Open Access Effect of Different Drying Methods on The Total Anthocyanin Content ofMangosteen (Garcinia mangostana L.) Rind Extract
Wijayanti, N. P. Ayu Dewi 1, Leliqia, N. P. Eka1, Setyawan, E. I.1, Widjaja, I. N. Kadjeng
1, Darayanthi, M. Y. Dwi1
1 Department of Pharmacy, Faculty of Mathematics and Natural Sciences,University of Udayana, Jalan Raya
Kampus Bukit, 80361, Denpasar, Bali, Indonesia
* Corresponding author. E-mail: dwijayanti27@gmail.com (A Author), leliqia@gmail.com (B Author) Sci Pharm. 201X; 8X: XXX–XXX doi:10.3797/scipharm.xxx
Published: Month DDth 201X Received: Month DDth 201X Accepted: Month DDth 201X
This article is available from: http://dx.doi.org/10.3797/scipharm.xxx
©Authoret al.; licensee ÖsterreichischeApotheker-Verlagsgesellschaft m. b. H., Vienna, Austria.
This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Abstract
Mangosteen (Garcinia mangostanaL.) rind contains phenolic compounds namely
anthocyanins that have the potential as a natural colorant. Drying extract containing anthocyanins requires a method that can be operated at low temperatures due to the thermolabile nature of the extract. The drying process of the extract can lead to reduction of components of the anthocyanin compounds as well as differences in the characteristics of the powder. In this research, the drying of mangosteen rind extract was carried out by using three types of drying methods namely freeze drying, fluidized bed drying, and oven drying so as to determine the effect of different drying methods on the total anthocyanin content.
This research is experimental in nature in which the total anthocyanin content of the extract before and after the drying process was measured by the pH-differential method using UV-Vis spectrophotometer. The data measured were the difference in value of total anthocyanin content before and after the drying process, followed by statistical analysis using the Kolmogorov-Smirnov and Levene tests to find out the distribution and homogeneity of the data; the data were then analyzed by the Kruskal-Wallis and Mann-Whitney tests with a confidence level of 95%.
The results of the research showed that total anthocyanin content of the extract can be affected by the difference in drying methods. The freeze drying method produced extract with the least reduction in total anthocyanin content compared to the other two methods. Of the three drying methods used, only the fluidized bed drying and freeze drying method which produced dry extract powder.
Keywords
Introduction
Mangosteen (Garcinia mangostana L.) rind contains phenolic compounds that have
the potential asa natural colorant [14, 7]. Anthocyanins are used as a colorant in a variety of food and beverage products [2]. Besides being able to be used as a colorant, anthocyanin also has some pharmacological activities such as anticarcinogenic and antiulcer [6,3]. Anthocyanin compounds in mangosteen rind are found to have antioxidant activity with the value of EC50 of 8.5539 µg/mL [15].
Anthocyanin content in mangosteen rind can be obtained by extracting it to obtain a liquid extract. Liquid extracts obtained from extracting must go through a drying process to obtain dry extract. According to journal [9], states that the dry extract showed some benefits compared to the liquid form, which include better stability and standardization and easier quality control. Drying the extract containing anthocyanin requires a method that can be operated at low temperatures because anthocyanin has a thermolabile characteristic. Several methods that can be used to dry the extract at low-temperatures are freeze drying, fluidized bed drying and drying by an oven.
Freeze drying is commonly used to dry the dosage form because it can increase the stability of thermolabile compounds during storage [12], while the oven is a dryer commonly used and can be set at low temperatures to maintain the stability of the material. Fluidized bed drying method can provide a dramatically higher heating capacity so as to increase the heat transfer to the masses and ease in the handling of drying products [13].
Drying the extract using different methods can affect the total content of anthocyanin of the extract. The drying process can lead to reduction of compound components of the dry extract compared to that of the fresh plant [8]. In this research, three methods of extract drying namely freeze drying, fluidized bed drying, and oven drying are used to determine the effect of different drying methods on the amount of total anthocyanins content of the extract.
Results and Discussion
A. Preparation of Crude Extract of Mangosteen Rind
Samples of mangosteen rind were extracted by maceration for 3 days using ethanol solvent acidified with citric acid 3% (w/v) where it was known that the composition of ethanol 70% extraction solvent with 3% citric acid was found to produce extract with the highest amount of anthocyanin. The resulting liquid extract had the organoleptic properties namely having the form of liquid, dark red color and smelled of alcohol.
B. Preliminary Test of Anthocyanin Content of Mangosteen Rind Extract
The extract was firstly tested using color and qualitative tests to know the presence of anthocyanin compounds which can be seen in Table 1.
Table 1.Result of Anthocyanin Preliminary Test
Testing Method Result of the
Added with the 2M NaOH
For the color test, acidic solution (HCl) and alkaline solution (NaOH) are used based on one of the factors that affect the color of anthocyanin which is pH. The acidic nature will cause the color of anthocyanin to turn into red, while alkalinity will cause anthocyanin to turn blue [10]. The qualitative test is carried out using the thin layer chromatography method by comparing the data ofthe retention value of the solute (Rf) which is not known and appropriate standard retention data (a known compound) under the same conditions [4]. Based on the color test using 2M HCl and 2M NaOH and the determination of the value of Rf using the thin layer chromatography (TLC) method, it was concluded that the sample contained anthocyanin compounds.
C. Extract Drying
The liquid extract obtained was divided into three parts, namely samples A, B, and C which was then dried using three different methods of drying where sample A was dried by using the freeze drying method, sample B was dried bythe fluidized bed drying method, and sample C was dried by a drying method using an oven.
Table 2.Organoleptic Observation Resultof Mangosteen Rind Extract
Sample Organoleptic Observation
SampleB : Dried by afluidized bed dryer
SampleC : Dried by an oven
[11]. The color of extract as a result of drying processis expected to be not much different from that of the extract before drying. The color of the extractas a result of different drying methods is different compared to the initial color of the extract which is dark red. Sample A has the color closest to its initial color. Change into brown of the color of the extract after a drying process indicates anthocyanin degradation as a result of heating. Samples A and C still have a distinctive smell of alcohol which indicates that there is still solvent content in the extract. The extract tastes bitter which may be caused by tannin content of mangosteen rind.
D. Determining the Total Anthocyanin Content (TAC) of the Liquid and Dry Extract
The total anthocyanin content (TAC) of the liquid extract (before drying) and dry extract (after drying) was determined by the pH differential method using a spectrophotometer. Anthocyanin compounds underwent reversible structural changes that resulted from changes in pH where the changes could be observed from the resulting absorbance difference [16]. Anthocyanin absorption dissolved in buffer at various pH conditions was measured at a wavelength of 510 nm which is the maximum wavelength of cyanidin 3-glycosides [15].
The measurement ofthe total anthocyanin content of the liquid extract sample and the extract sample after being dried was done repeatedly three times. The result of measurements of the total anthocyanin content (TAC) can be seen in Figure 2.
Figure 1. Result of Measurement of the total Anthocyanin Content (TAC) of the Mangosteen Rind Extract Before and After Drying
From the result of the measurement, the extract dried by the freeze drying method produced the highest amount of total anthocyanin content compared tothe fluidized bed drying and ovendrying methods. From the overall results, the amount of anthocyanin decreases if the extract is dried. According to journal [8], extract drying process can lead to reduction of compound components in dry extract compared to that of the fresh plant.
In order to determine the effect of different drying methods on the total anthocyanin content of the extract, the data on the difference in the total anthocyanin contentresulting from the three methods of drying were analyzed statistically. The difference in the total anthocyanin content of the extract before and after drying was statistically analyzed by first
of all looking at the distribution and homogeneity of the data using the Shapiro-Wilk test and Levene test. From the result of the Shapiro-Wilk test, it was found out that the data distribution was not normal where the value of (P) <0.05 was significant. Therefore, the statistical analysis was followed by the Kruskal-Wallis non-parametric analysis (with a confidence level of 95%) to demonstrate statistically whether there was any effect of different extract drying methods on the difference between the total anthocyanin content of the extract after drying and followed by the Mann-Whitney test to clarify the difference in each drying method.
From the result of Kruskal-Wallis analysis, the significant value of P <0.05 obtained indicates that different drying methods have an effect on the difference in the total anthocyanin content of the extract. Further test using the Mann-Whitney test showed that the three applied methods of drying gave the effect of a significant difference in the value of the total anthocyanin content of the extract namely P <0.05. The result of the statistical analysis showed that the freeze drying method was able to provide the least and different reductionin the amount of total anthocyanin content compared to the other two methods. This was because the temperature used was very low (-15 °C) to dry the extract. Meanwhile, the fluidized bed drying method and oven used a highertemperature of 35 ºC.
Based on this, it can be said that the difference in drying methods have an effect on the total anthocyanin content of the extract of mangosteen rind in which drying with the freeze drying method gives the smallest decrease of total anthocyanin content of the extract compared to the fluidized bed drying method and oven drying method. Of the three drying methods used, only the fluidized bed drying and freeze drying method which produced dry extract powder.
Experimental A. Materials
Materials used in this study is the mangosteen rind, citric acid (Brataco), distilled water (Brataco), Ethanol 70% (technical, Brataco), HCl (p.a., Merck), NaOH (p.a., Merck), butanol (p.a., Merck ), concentrated acetic acid (p.a., Merck), NaCl (p.a., Merck), CH3COONa.3H2O (p.a., Merck), KCl (p.a., Merck).
B. Preparation of Mangosteen Rind Samples
Mangosteen fruits were obtained from Luwus Village, Sub-district of Baturiti, Tabanan Regency, Bali. Determination of plants was done at UPT Balai Konservasi Tumbuhan (Plant Conservation Center) of Kebun Eka Karya Bedugul, Tabanan, Bali.
The collected mangosteen fruitswereeach split into two and their flesh was removed. The rind was then sorted wet under running water then drained and aerated briefly. The mangosteen rind was then cut into small pieces.
C. Preparation of Crude Extract of Mangosteen Rind
D. Tests on the Mangosteen Rind Extract 1. Preliminary Test
The preliminary test was conducted to determine the presence or absence of anthocyanin compounds in the sample. The test was carried out on the crude extract, liquid extract, and dry extract of each drying method. The testwas carried out with the color test method and Thin Layer Chromatography test method.
- Color Test
The color test was carried out by adding to the sample HCl heated and the result is positive if it the red color appears. NaOH was also added to the sample while observing the color changes which occurred. The result is positive if the slowly fading green-blue color appears [15]
- Thin Layer Chromatography Test
The extract was spotted on a TLC plate of silica gel GF 254 and eluted by using different mobile phase, i.e.HCl 1% and BAA (butanol: acetic acid: water at the ratio of 4: 1: 5 v/v). The given Rf value was calculated. The Rf value of anthocyanin in the mobile phase ofHCl 1% is in the range of low to mid and the Rf value of anthocyanin in the mobile phase of BAA is moderate (0.10 to 0.40) [15].
E. Extract Drying
Liquid extract of mangosteen rind was divided into three parts. One part was dried using a freeze dryer at a temperature of -15 ° C with a pressure of 0.5 atm. The dry extract obtained was then weighed. Another part was dried using a fluidized bed dryer with the addition of lactose to obtain dry extract. The dry extract obtained was then weighed. The other one partwas dried using a vacuum oven at a temperature of 30ºC and a pressure of 0.5 atm. The dry extract obtained was then weighed.
F. Measuring the Total Anthocyanin Content
The total anthocyanin content was measured using the pH differential method. Measurement was made to the condensed extract and powder extract of mangosteen rind. The sample was dissolved in the sodium chloride-HCl buffer with the pH of 1.0 and sodium acetate trihydrate buffer with the pH of 4.5. The absorbance of the solution was measured at a wavelength of 510 nm. Sample dilution factor is determined by dissolving the sample inKCl buffer with the pH of 1.0 until the absorbance at a wavelength of 510 nm reaches 1.2. The sample was then dissolved in KCl buffer with the pH of 1.0 (allowed to stand for 15 minutes) and in CH3COONa.3H2O buffer with the pH of 4.5 (allowed to stand for 5 minutes). The absorbance of the solution was then read at a wavelength of 510 nm and 700 nm. The final absorbance was calculated by the formula: A = (A510-A700)pH 1,0 – (A510
-A700)pH 4,5. [16]. Total anthocyanin content or was calculated by the following formula:
Total anthocyanin content (mg/L) =
This study used the following formula [1]:
Total anthocyanin content (mg/100 g sample) =
Description:
A : (A510-A700)pH1,0– (A510-A700)pH 4,5
DF : Dilution factor
V : The final volume or the volume of the sample after dilution Wt : Weight of the extract (g)
l : Width of the cuvette (1 cm)
G. Data Analysis
The data obtained from the measurement were statistically analyzed using the Kolmogorov-Smirnov test to see the distribution of data and Levene test to see the homogeneity of the data. If the distribution of data is normal and homogeneous, the data analysis is followed by a unidirectional analysis namely Analysis of Variance (ANOVA) one-way, with a 95% confidence level. Furthermore, in order to identify which of the drying methods that gives the effect, the statistical testis followed by LSD test.
Acknowledgement
We would like to express our sincere gratitude to LPPM of University of Udayana, Laboratorium of Technology of Pharmacy Department, Faculty of Mathematics and Natural Science of University of Udayana and Bandung Institute of Technology.
Supporting Information
Author(s)’ Statement(s)
Competing Interests
The author hereby state that this research has no conflict of interest.
Informed Consent, Ethical Approvals
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Animal Rights
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