The Efficacy of Mangosteen (Garcinia Mangostana) Peel Extract As An Alternative Stain To Eosin in Wright'S Stain

THE EFFICACY OF MANGOSTEEN
(Garcinia mangostana)
PEEL EXTRACT AS AN ALTERNATIVE
STAIN TO EOSIN IN WRIGHT’S STAIN
Background of the Study
 Mangosteen peels contain a number of pigments that comes from two metabolites,
namely mangosteen and β- mangosteen. If all of the content contained in the
mangosteen fruit is extracted, will be obtained natural dye in the form of
anthocyanins which generally produces red, purple, and blue colors. Mangosteen
peels also contain flavan-3,4-diols, which classified as tannin compounds and can
be used as natural dyes on fabric. The dyeing result using mangosteen peels material
showed that each color intensity generated from the fixation process using iron (II)
sulfate, alum and lime will be progressively increased in line with the increasing of
the washing time applied. (Kusumawati N., et al., 2017).
 The major bioactive compounds found in Mangosteen are phenolic acid,
prenylatedxanthone derivatives, anthocyanins, and procyanidins. Ten phenolic acids
were identified in Mangosteen fruit. The major anthocyanin in mangosteen was
cyanidin-3-sophoroside (Chaovanalikit, et al., 2012).
Background of the Study
 The physico-chemical properties such as moisture content, ash content, total solids
content and pH of Mangosteen peel were determined by Association of Official
Analytical Chemists (AOAC) method. The chemical compounds present in
Mangosteen peel were investigated by phytochemical tests. Natural dye was extracted
from Mangosteen peel with pure water at 70ºC for 60 minutes. (Universities Research
Journal 2016)
 The chemical compounds responsible for the indicator properties of the mangosteen
is called anthocyanin that gives Garcinia mangostana its purple color characteristic.
Anthocyanin is the major pigment responsible for the colouration with dye ranging
from orange, pink, red, violet to blue in the flowers and fruits. The purplish/reddish
extract of this fruit may have a staining effect on a thin blood smear in clinical
laboratory practice that may be used as an alternative organic stain.
Statement of the Problem
The conduct of this study aimed to examine the the efficacy of
mangosteen peel extract as an alternative stain to Eosin in Wright’s
stain. Specifically, this study answered the following questions:
1.) What are the effects of the different concentrations of mangosteen
in staining blood smears in terms of the following concentrations;
15 grams in 100 mL ethanol;
20 grams in 100 mL ethanol;
25 grams in 100 mL ethanol.
Statement of the Problem
2. What is the effect of the different concentrations mangosteen peel
extract as in terms of:
(a) visual clarity;
(b) cell morphology demonstration;
(c) uniformity of staining.
3. Is there significant difference between the mangosteen peel extract

concentrations with the control, Wright’s stain.
Significance of the Study
The findings of this study will redound to the benefit of the society
considering that dyes play an important role in peripheral blood smear
staining. The significance of this study are the following:
 To observe the staining capabilities and efficiency of dye from the

Mangosteen peel in the blood film because natural dye is easy to
extract, non-toxic and have the antimicrobial properties.
Significance of the Study
 Economical rate of mangosteen peel will be increased because of
the demands of this fruit. Therefore, the utilization of mangosteen
peel as raw material for natural dye, which focuses on the
regeneration of waste and eco-friendly materials will be recognized.
 To prove the cost efficiency of the mangosteen peel extract
compared to the commercially available Eosin reagent used in
peripheral blood staining.
METHODOLOG
Y
Preparation of Materials
 Magosteen (Garcinia magostana) utilized in this study were
obtained from the local market of City of Koronadal, South
Cotabato, Philippines. While in obtaining blood samples from 15
students of Notre Dame of Marbel Univeristy, needle and syringe
method of venipuncture were used. The whole experiment was
conducted inside the Notre Dame of Marbel University Biology and
Chemistry Laboratory wherein the staining and the extraction
process will occur.
Preparation of Mangosteen peel stain
 The mangosteen peel were placed in a clean plastic container with sealed lid
and stored in room temperature for 24-48 hours before being processed.
Extraction and preparation of the solution were done in line with the study
of Chukwu (2011) and Adegoke et al. (2010) where the mangosteen peel
was washed and dried under the shade for minimum of 7 days and then
milled to form the fine powder. The powder will be stored in glass jars and
refrigerated at 4°C until require for use. Fifteen (15), twenty (20), and
twenty-five (25) grams of the powder was weighed using digital balance,
which was dissolved in 100 mL of ethanol in a beaker and then filtered
using the filter paper. It was transferred to 5mL test tubes, which were
centrifuged at 3000 rpm for 5 minutes and supernatant were collected in a
coupling jar with the use of micro pipettes.
Staining of Blood Smears
 The blood colleced from the 15 respondents must be anticoagulated preferably with
the disodium or dipotassium salt of ethylene diamine tetra acetic acid (EDTA).
Staining is done by placing 0.05 mL (50 uL) of blood on a slide and spread lightly to
rapidly produce a smear. The smea The slide was fixed in the methanol for 5 minutes
and dry. Methylene blue is place upon the smear for 2 minutes and then rinse with
running water. Eosin Y is added on the smear for 1 minute. Then, the slide is rinse
again with water and let to dry (Abnova, 2010). All steps are repeated in another slide
except that the mangosteen peel extract is used in exchange of Eosin Y and for each
blood sample. Four smears were made sorted according to the following groups:
Group I (Control Group): Stained with Wright’s stain, Group II: Stained with 15
grams in 100 mL ethanol mangosteen extract, Group III: Stained with 20 grams in 100
mL ethanol mangosteen extract, and Group IV: Stained with 25 grams in 100 mL
ethanol mangosteen extract.
Comparison of the staining effects
 Stained smears are graded as:
 (1) very poor (no observable morphology of cells);
 (2) poor (stained red blood cell only, morphological alterations present);
 (3) satisfactory/average (visible stained red blood cells, platelets with
some morphological alterations);
 (4) good (visibly stained red blood cell, platelets with no morphological
alterations) and
 (5) very good (visibly stained red blood cells, platelets and white blood
cells with no morphological alterations).
Insights
 The context of this research was to show the staining quality and capacity of the Mangosteen
peel extract as an alternative stain to Eosin Y in Wright's stain.
 As we were writing this research paper, we had the difficulty in communicating to each other as
we are in our new normal situation. It is quite difficult to share our ideas through video calls and
chats because of the limited features and as well as the connectivity to the internet.
 We also had to revise our title from one to another in order to come up with a good, easy to
understand title.
 In searching for our related literatures, we also had a difficulty as there were only a few
literatures about the mangosteen extract as an alternative stain in peripheral blood staining.
However, there was a study that concerns about the Mangosteen fruit extract as an alternative
for Histology staining. We incorporated that study in our research to get recommendations and
ideas.
 Despite of all these struggles, we managed to overcome it with the help of the seminars
conducted through online meeting. We also want to thank our advisers for giving us their time
and effort to help us to make our research paper better.
THANK YOU 

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THE EFFICACY OF MANGOSTEEN

3. Is there significant difference between the mangosteen peel extract

 To observe the staining capabilities and efficiency of dye from the

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