Extraction, Identification, and Gel Formulation of Mangiferin from

Mango (Mangifera indica L.) Leaves Extract

Rudi Afrinanda, Yusa Ristiawati, Muhammad Shoufi Islami, Deasy Vanda Pertiwi

Faculty of Pharmacy, Ahmad Dahlan University

Keywords: Mangiferin, Mangifera indica, gel, wound healer, ulcus diabetic

Abstract: Mango (Mangifera indica L.) leaves contain flavonoid which has anti-inflammatory and antioxidants effects

that are beneficial on healing diabetic ulcers. The extract made in gel formulation because it was easy to

dry, forming the washable film layer that provides a cool sensation on the skin. Gel components influence

the stability of formula. To ensure gel quality, safety, and benefits, physical stability test was needed to

fulfill the specifications and stability during storage. This study aimed to extract and identify the mangiferin

as an active compound in mango leaves and to formulate Mangifera indica leaves extract gel as a wound

healer. Extraction of Mangifera indica leaves used soxhlet method with ethanol 70% and determined using

TLC-densitometry method. The optimum formula of the gel was determined by variations of CMC-Na

concentration as gel base, and the compliance of the gel characteristics. The analysis of characteristics

included spreadability test, homogeneity test, adhesivity test, and pH test. The result of extraction was

determined by TLC-Densitometry as 330,52 mg/gram of viscous extract. The formula with 5% CMC-Na gel

base complied with the required characteristics and was the optimum formula, which stability analysis did

not show any changes in pH, colour, consistency, adhesivity and spreadability during storage.

1 INTRODUCTION

Diabetes Militus (DM) is a disease characterized by

the occurrence of hyperglycemia and carbohydrate,

fat, protein metabolism disorders, as a result of

disorder or insulin deficiency by β Langerhans cells

of the pancreas gland, or caused by the lack of

responsiveness of body cells to insulin. One of the

complications of DM occurring is diabetic ulcers or

diabetic lesions, which are skin lesions caused by

high blood glucose levels resulting in vascular

resuscitation and further vascular neuropathy

(Fatimah, 2015, Sarwono, 2009). Based on the data

of the Indonesian Ministry of Health (2014), the

prevalence of diabetic ulcer wounds in Indonesia

reaches 54%. This disease is often found in

developing countries; Indonesia was ranked seventh

with a number of 10 million diabetic patients in

2015 (IDF, 2015).

Using antioxidants as a treatment on diabetic

wounds is the most effective approach related to

wound healing of diabetes. One of the types of

plants that is potential as a wound healer of diabetes

is mango (Mangifera indica). Mango leaves contain

active compound of mangiferin that acts as

antioxidant and capable of lowering blood sugar

levels in diabetes therapy. Moreover, extracts of

mangiferin has a potential for the healing of wounds

in diabetes (Fithriyani et al., 2014, Khandare, 2016).

Mangiferin total from ethanol extracts of Mangifera

indica is 102 mg/gram of mangiferin compounds.

This plant grows a lot in the community and only the

fruit are commonly consumed, not yet optimally

utilized in increasing the value of use (Fithriyani et

al., 2014).

Utilization in the community is seen as not

optimal yet because it has not been processed into

useful drugs; therefore, it needs formulations to form

products, i.e. preparations in the form of a gel. The

gel is a semi-solid material consisting of a

suspension made of inorganic particles that are small

or large organic molecules including penetration by

a liquid. The gel preparation is chosen because it is

easy to dry out, forming a layer of film that is easily

washable and provides a cool sensation on the skin

(Ansel, 2008, Panjaitan et al., 2012).

Formulation of gel in this study used CMC-Na as

the gel agent. CMC-Na is a polymer derivate

cellulose that quickly expands when supplied with

hot water and neutral, clear crystal and has a strong

138

Afrinanda, R., Ristiawati, Y., Islami, M. and Pertiwi, D.

Extraction, Identiﬁcation, and Gel Formulation of Mangiferin from Mango (Mangifera indica L.) Leaves Extract.

DOI: 10.5220/0008240701380142

In Proceedings of the 1st Muhammadiyah International Conference on Health and Pharmaceutical Development (MICH-PhD 2018), pages 138-142

ISBN: 978-989-758-349-0

bond between molecules (Aponno et al., 2014). In

this study, the variation of gel base CMC-Na was

analyzed to find the optimum formula. It is specified

based on gel characterized, i.e. spreading test,

adhesion test, homogeneity, consistency, and pH.

CMC-Na has advantages over Carbopol; pH of

CMC Na is higher than carbopol, the spreading

power of CMC-Na is greater than carbopol gel, and

also the extraction into CMC-Na does not affect the

spreadability, while the gel of carbopol decreased

power of scatterplot (Maulina and Suhigartini,

2015).

2 MATERIALS AND METHOD

2.1 Materials

Mango leaves (Mangifera indica), 70% Ethanol,

ethyl acetate, glacial acetic acid, formic acid,

methanol, CMC Na, Tragakan, propilenglycol,

Carbopol, glycerin, Methyl paraben, and Aquadest.

2.2 Methods

2.2.1 Preparation of Ethanolic Extract of

Mango Leaves

Mango leaves from the area of Sleman, Yogyakarta

were picked and dried under the blazing sun and

previously washed beforehand; they were covered

with black cloth in the process to avoid direct

contact with sun rays. To obtain even drying, leaves

were then moved into oven for 2-3 hours at a

temperature of 500-600 °C, before being ground to

make powder leaves and sieved with sieve mesh no.

40.

About 625 grams of leaves powder was

transferred into a soxhlet tool and then added with

1500 mL ethanol as solvent. Extraction was

performed for 48 hours (Sachin et al., 2014). The

extract obtained was collected and concentrated on

evaporator to evaporate in a waterbath until viscous

extract was obtained.

 =

ℎ  

ℎ  

100%

(1)

2.2.2 Preparations and Determination of the

Optimum Formula of Mango Leaf

Extract Gel

Mango leaves extract (MLE) gel was formed from

MLE and excipients; the composition of the gel

formulated by a trial-error method in the

preformulation step. According to Adnan (2016),

MLE gel with CMC-Na as gel base has the

composition as in Table 1.

These formulas compared and evaluated to

choose the optimum one. The evaluation includes

organoleptic test and homogeneity, consistency, pH,

adhesive test and spreading test. The results obtained

are indicated in Table 2.

2.2.3 Organoleptic Test and Homogeneity

The organoleptic test was performed by directly

observing the colour and smell. Homogeneity test

carried out by applying the gel on a piece of glass

(Maulina and Sugihartini, 2015).

Table 1: Formula of mango leaves extract (MLE)

gel.

Component F1 F2 F3

Mango leaf extract

(MLE)

4% 4% 4%

CMC-Na 5% 5% 5%

Gliserin 5% 5% 5%

Pro

ilen

likol 2,5 % 2,5 % 2,5 %

in 0,25% 0,25% 0,25%

Aquadest ad 50

grams

Table 2: Physical evaluation of MLE gel.

Evaluation Concentration

FI F2 F3

anole

tic Colou

Ver

thick brownish

reen Thick brownish

reen

rownish

reen

Smell Ver

stron

Stron

Less

Homogeneity Homogenous Homogenous Homogenous

H 5 5 5

eadibility 8,117 c

6,4 cm 4,13 c

Adhesivit

3,4 secon

5,4 secon

56,7 secon

Consistenc

Stable Stable Stable

Extraction, Identiﬁcation, and Gel Formulation of Mangiferin from Mango (Mangifera indica L.) Leaves Extract

139

2.2.4 pH Testing

pH test was done using pH paper universal that was

dipped into the diluted samples. Colour change on

the pH paper was compared with the universal

standard (Maulina and Sugihartini, 2015).

2.2.5 Spreadability Test

About 0.5 grams of gel was placed on the glass and

another round glass was placed on top of it, and left

for 1 minute. After that, 150 grams of weight was

added and left for 1 minute. The diameter was then

measured (Astuti et al., 2010).

2.2.6 Adhesivity Test

About 0.25 grams of sample was placed between

two glass objects; given a load of 1 kg for 5 seconds,

which was then lifted and changed with 80 gram of

weight. The time of the release of the gel determined

as adhesivity of gel (Miranti, 2009).

2.2.7 Consistency Test

Consistency test was done using centrifugation test.

Gel samples were centrifuged at 3000 rpm for 5

minutes, and then observed of physical changes

(Djajadisastra et al., 2009).

2.2.8 Data Analysis

The data obtained was processed in statistics using

SPSS 24 software. Normality test (Shapiro-Wilk)

and homogeneity test (Levene) were performed on

the data. To see the relationship between the

treatment groups, the variations were analyzed in

one way (ANOVA) if the data was distributed

normally and homogenously. If the data was not

normal, Gaussian and then Kruskal-Wallis analysis

were done.

3 RESULTS AND DISCUSSION

Mashed dried mango leaves formed the smooth

brownish-green colour with a distinctive odour. Ten

pounds of dry mango

leaves produced 0.8

kilograms of powder. The water content in the dried

mango leaves was 8% ± 0.5, which met the standard

IE (<10%). If water content >10%, it can cause the

onset of enzymatic processes and cause damage due

to microbes, which can change the chemical content

of it.

Mango leaf extract obtained by the soxhlet

method, using ethanol 70% as the solvent, was left

for 24 hours and the ethanol 70% was vaporized on

the waterbath to obtain extracts. The extract

obtained had a bitter taste and a distinctive smell.

Yield and extract obtained was 12.5%.

The identification of the xanthan group carried

out was qualitative and quantitative. Qualitative

analysis was done by FeCl3 5% and HCl 1M. The

colour that was changing to brownish blackish

colour proved that the extract contained compound

of the xanthan (mangiferin), positive. Quantitative

analysis was done with the Densitometry method to

obtain TLC mangiferin levels using the stationary

phase of GF 254 and mobile Phase Chloroform:

Methanol: comparison with Formiat Acid 90:10:3.

Test results are presented in Figure 1.

TLC densitometry analysis aimed to determine

levels of mangiferin and quite economical since it

used relatively little motion phase and relatively

TLC densitometry analysis aimed to determine

levels of mangiferin and quite economical since it

used relatively little motion phase and relatively

short time and the measurement of levels of samples

simultaneously. From the analysis, the obtained

concentration of mangiferin was 330.52 mg/gram of

extract.

In the formulation gel, MLE was the active

ingredient. CMC-Na served as gel base, while

propilenglycol and Glycerin as humectant that

increased the stability of formula. CMC-Na was

used as the gel base in the formula because it has

good stability in acid and alkaline condition (pH 2-

10). Propilenglikol in gel formula was used as a

humectant to maintain the stability while keeping the

moisture content in the material properties of the gel.

This material can be stable at pH 3-6. The most

influential factors in the physical quality of the gel

preparations are the base and humectant. The base

gel will form a structure which is an important factor

in the gel formulation. Humectant serves to keep the

Figure 1: TLC plate of MLE in UV 245.

MICH-PhD 2018 - 1st Muhammadiyah International Conference on Health and Pharmaceutical Development

140

Figure 2: pH diagram with variations of the base

formula of CMC Na 2,5%% (FI), 5% (FII), dan 7,5%

(FIII).

gel formulation by absorbing moisture and reduce

evaporation of water from the formulation. Methyl

paraben was used as a preservative because gel has a

high moisture content that can lead to the occurrence

of microbial contamination.

The physical properties test of the gel are

required to guarantee the quality of gel formulation.

Test results from the third formula shown in the

Table 2. pH of topical formulation requirement is 5-

7 because the normal pH skin IE 4.5 – 6.5

(Martin,1983). Test results are presented in Figure 2.

The gel in this research has pH of 5, which is

appropriate for skin pH. If the pH is not a match for

the skin, it will irritate the skin and reduce comfort

when applied to the skin.

Spreading power test done to learn the ability of

the gel to spread on the skin. The terms of the power

of spread in the topical formulation are 5-7 cm

(Ulaen 2012, et al.). Increasing spreadability of

topical formulation will simplify in the application.

Figure 3 presented the test results.

In this research, the spreading power that can

qualify in specified. The higher concentration of gel

base will cause the smaller area of spreading. It is

because in the high concentration of base gel, it

contains the lower of water. It makes increasingly

viscous gel consistency so that the spreading power

decline. Statistic analysis for the third formula, there

are F1, FII and FIII, normal distributed data and

homogeneity of data that show a different result. It is

indicated by one way ANOVA statistics with value

p = 0,200 or > 5% of the value that is assigned. So

that the third formula meets the requirements.

To see the ability of the gel in attaching to the

skin, adhesivity test was performed. Power

requirement for the material to latch onto the skin is

no shorter than 4 seconds (Ulaen, et al., 2012).

Figure 4 presented the test results.

The formulas that met the requirements were

formula II and III. It is because the lower

consistency made shorter time in contact with skin.

In addition, the increase of concentration caused a

thick consistency that also was increasingly

adhesive. Statistic analysis of normal distributed and

homogeneity by ANOVA retrieved result of the sig

(5%; p < p = 0.00) and the data is not homogeneous

(5%; p < p = 0.04), so then the Kruskal Wallis test

produced a significant value of (p 5%; < p = 0,026).

It means the difference of concentration affected

adhesivity of the gel. Consistency test showed that

segregation in gel formulation did not occur. It

indicated that the gel formula were stable in storage.

Figure 3: Spreadability diagram with variations of the

base formula of CMC Na 2,5 %(FI), 5% (FII), dan 7,5%

(FIII).

Figure 4: Adhesivity diagram with variations of the base

formula of CMC Na 2,5 %(FI), 5% (FII), 7,5% (FIII).

Extraction, Identiﬁcation, and Gel Formulation of Mangiferin from Mango (Mangifera indica L.) Leaves Extract

141

4 CONCLUSION

The results showed the MLE gel formulation were

green-brown with a distinctive smell, homogeneous,

pH 5, spreading 4.13-8.117 cm, latching for 3.4-56.7

seconds, and stable in storage. The most optimal

formula is gel with a concentration of CMC Na 5%.

ACKNOWLEDGEMENT

The authors would like to thank The Ministry of

Research Technology and Higher Education for

providing the grant used in this study.

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