ACTA SCIENTIFIC NUTRITIONAL HEALTH

Volume 3 Issue 9 September 2019

Research Article

Total Phenolic Content of Mango Pulp (Mangifera indica L.) Extracted using
Three Different Solvents

Jemima Beryl Mohankumar*, L Uthira and S Uma Maheswari

PSG College of Arts and Science, Coimbatore, India
*Corresponding Author: Jemima Beryl Mohankumar, PSG College of Arts and Science, Coimbatore, India.
Received: July 02, 2019; Published: August 22, 2019

DOI: 10.31080/ASNH.2019.03.0421

Abstract
Mango is probably the most variegated fruit of India. Mango fruit is not only rich in macro and micro nutrients, but also rich in
natural antioxidants. Phytochemical and nutrient content appears to vary across mango cultivars. Seven varieties of mango available
were purchased from the local market by following a sampling plan. The pulp was recovered, blended and used for further extraction
and analysis. Three solvents namely ethanol, methanol and water were used for extraction. Total phenolic content was determined by
the Folin-Ciocalteu method. The TPC of mango varieties were from 111.73 ± 3.56 to 889.72 ± 15.73 µg of GAE/g FW. The results were
superior in water extracts, medium in ethanol and least in methanol in all the mango varieties except Alphonsa mango. Quantification
of total phenolic components would be used to evaluate antioxidant activities

Keywords: Mango Varieties; Solvent Extraction; Total Phenols; Folin-Ciocalteu; Mango Pulp

Introduction showed that those major antioxidants (i.e., vitamins, flavonoids, ca-
Mangifera indica L. (mango), “the king of fruits” belonging to rotenoids, polyphenols, mangiferin, etc.) contribute directly to the
the family Anacardiaceae, is one of the most popular fruits in tropi- total antioxidant capacity of mango [3-5].
cal regions. Mango is probably the most variegated fruit of India. In one study five varieties of mangoes from four countries were
There are almost 500-1000 cultivars of different mangoes in the evaluated with multiple harvests over one year to compare the
country. This huge variety not only differs in its shape, size and co- beta-carotene, ascorbic acid, and total phenolic contents and an-
lour but also in its scrumptious taste. They are seasonal, available tioxidant capacities of the fruit pulp and to compare the phenolic
from April to July. However it is not possible to obtain all varieties profiles of the individual varieties. Phytochemical and nutrient con-
in one particular place. tent appeared to vary across mango cultivars. Up to 25 different ca-
Databases on mango varieties list the most number of varieties rotenoids were isolated from the mango pulp, the densest of which
from India. To name them viz. Alphonso, 'Banganapalli, 'Bombay, was beta-carotene, which accounted for the yellow-orange pigmen-
'Bombay Green, 'Chausa, 'Dashehari, Fazli, Fernandian, Himsagar, tation of most mango cultivars. Total phenolic content ranged from
Kesar, Kishen Bhog, Langra, Mallika, Mankurad, Mulgoa, Neelum, 19.5 to 166.7 mg of gallic acid equivalents (GAE)/100 g of puree.
Pairi, Samar Behisht Chausa, 'Suvarnarekha, 'Totapuri, Vanraj, Zar- The varieties Tommy Atkins, Kent, Keitt, and Haden had similar to-
dalu, Amrapali, Bangalora, Gulabkhas in all more than 25 varieties tal phenolic contents, averaging 31.2+/-7.8 mg GAE/100 g of puree,
[1]. whereas the variety Ataulfo contained substantially higher values.
Similar trends were observed in the DPPH radical scavenging activ-
Mango is one of the most important fruits in terms of produc- ities among the five varieties. In contrast, the country of origin and
tion, marketing, and consumption. Among tropical fruits, it ranks harvest dates had far less influence on these parameters. Ataulfo
second, next to banana in international trade. Mango fruit is not mangoes contained significantly higher amounts of mangiferin and
only rich in macro and micro nutrients, but also rich in natural an- ellagic acid than the other four varieties. Large fruit-to-fruit varia-
tioxidants. Mango had a higher content of ascorbic acid, which was tions in the concentrations of these compounds occurred within
nine and four times that of the apple and banana, respectively [2]. sets of mangoes of the same cultivar with the same harvest location
Similarly, its carotenoid content also held a significant superior- and date [3].
ity, with nearly 10 times that of the apple or orange. α-tocopherol
was another preponderant composition, which was more than 18 Some of the varieties available in the present study area are de-
times that of the apple and outdistances that of the grape, orange, scribed below
tangerine, and banana. Moreover, many previous studies have

Citation: Jemima Beryl Mohankumar., et al. “Total Phenolic Content of Mango Pulp (Mangifera indica L.) Extracted using Three Different Solvents". Acta
Scientific Nutritional Health 3.9 (2019): 165-168.
Total Phenolic Content of Mango Pulp (Mangifera indica L.) Extracted using Three Different Solvents

166

• Alphonso: Also known as Hapoo, this is the most expensive stem and seeds.
variety of mangoes in India. Maharashtra, Gujarat and Karna-
The seven types of cultivars of ripe mangoes (Alphonso, Sen-
taka are its prime producers. It has a fibreless pulp and dous-
thuran, Malgova, Nadusalai, Emmabasanth, Banganapalli and Kud-
es into the mouth as a smooth-creamy mango. This variety of
hadath) were purchased from local market in Coimbatore. The
mango is popular all over the world.
mangoes were washed with warm water and peels were removed
• Banganapalli: These mangoes have a very beautiful yellow by using sharp stainless steel knife and underlying pulp was re-
colour and are sweet and fibreless. It is the most popular va- covered by scraping with the blunt edge of the knife. The pulp was
riety of mangoes which comes straight from Andhra Pradesh. ground using mixer and grinder (Preethi Chef Pro-750W) individu-
• Neelam: This is one variety that grows throughout the coun- ally up to the formation of a coarse texture.
try. With its specialty in Hyderabad, it is a large-yielding vari-
The blended sample (0.3 g) was weighed using an analytical bal-
ety of mangoes. Though it arrives early in the season, the best
ance (2007 TX/TXB Series, Shimadzu). The sample was then trans-
variety is available only in June.
ferred into a Borosil test tube. For each test tube, 6 ml of extracting
• Sindoora: Its reddish color at the top has entitled this vari- solvent was added to the sample (solvent ratio of 1:20). The test
ety with the name of Sindoora. Extremely juicy and pulpy, this tube was then placed on a water bath shaker (Precision) of 130
mango is one of the tastiest mangoes one can ever have. rpm at a room temperature of 30ºC for 60 min for the extraction.
• Malgova: Mammoth in size, the Malgova is legendary to Hy- The extracts were collected and filtered using a syringe filter be-
der Ali’s Mysuru orchards, where each fruit weighed between fore the determination of total phenolic content. All the extractions
1-1.5 kg a piece! When compared to the waif-like Alphonso, were carried out in three replicates. Extracts were stored in brown
the Malgova almost looks unsightly. The flavour though, is a bottles in the freezer until further analyses.
welcoming sweetness with the slightest undercurrent of tart-
Polyphenols are isolated by the three steps of extraction, incu-
ness. The pink blush at the tip is the only indicator of its ripe-
bation and filtration of the supernatant. It is widely accepted that
ness.
the extraction step is one of the most important stages in isolation
• Imam Pasand: This variety originated Andhra Pradesh. This of polyphenols, but based on literature, there is no consensus about
variety is also called Imam Pasand. Himayat is popular among one single and effective standard extraction method. On the con-
the Nawabs. Each weighs around 450g-500g and hence each trary, there are several reported methods with very accurate re-
fruit is large. Ripened fruit has negligible amounts of dietary sults, and according to the literature in some cases, the solid-liquid
fibre and has a silky flesh with deep sweet flavour and distinc- extraction with different types of solvents is more adequate [6].
tive citrus overtones. The skin is green and when it ripens, it
attains yellow colour. In order to extract different phenolic compounds from plants
with a high degree of accuracy, three solvents of differing polarities
were used. Solvents used for the extraction of bio molecules from
Materials and Methods
plants are chosen based on the polarity of the solute of interest. A
Sampling and extraction
solvent of similar polarity to the solute will properly dissolve the
The mango fruit samples were purchased from retail outlets in solute. Multiple solvents can be used sequentially in order to limit
the Coimbatore area and prepared for analysis during the harvest the amount of analogous compounds in the desired yield. How-
time. The retail outlets included supermarkets, independent retail- ever, in our study we used three different solvents for extraction
ers and catering suppliers. One kilogram each was purchased at viz., ethanol, methanol and deionised water. Variations in the yields
least from three outlets and was combined into composite samples and phenolic contents of various extracts are attributed to polari-
for analysis. Each composite was made up of three sub-samples, ties of different compounds present in the sample. A higher content
combined on an equal weight basis. Sub-samples included were of polyphenols can be obtained with an increase in the polarity of
based on the need to take into account factors including cultivar the solvent used. Several authors have reported a good correlation
and region. From this the laboratory sample and test sample were between total phenolic contents and reducing power (r 2 = 0.86)
derived for the determination of TP. and radical scavenging activity (r 2 = 0.82) assays. The polarity of
solvents in the increasing order is as follows: Hexane < Chloroform
The fresh fruits were sampled in seasons (summer) where the
< Ethylacetate < Acetone < Methanol < Water [7].
cultivars and geographic origin were known to change between
seasons. This process allows a single, robust set of nutrient values
Chemicals
to be derived for each fruit. A voucher specimen was identified by
at least three persons viz., the investigator, the vender or farmer Gallic acid was purchased from Sigma-Aldrich USA. Folin- Cio-
and botanist deposited at the Department’s Nutrition laboratory calteu reagent was obtained from Merck, Germany. All other chem-
for analyses and future reference. The samples were stored in the icals used in the study were of analytical grade. These were sup-
refrigerator and extracted the same day after removing the skin, plied by the local agent in the study area.

Citation: Jemima Beryl Mohankumar., et al. “Total Phenolic Content of Mango Pulp (Mangifera indica L.) Extracted using Three Different Solvents". Acta
Scientific Nutritional Health 3.9 (2019): 165-168.
Total Phenolic Content of Mango Pulp (Mangifera indica L.) Extracted using Three Different Solvents

167

Determination of total phenolic content ricultural patterns. Further research is needed to find out the vari-
The total phenolic content of the mango extract was deter- ous factors influencing phenolic content in plants.
mined by using Folin-Ciocalteu reagent following a slightly modi-
The TPC of mango varieties were from 111.73 ± 3.56 to 889.72 ±
fied method [11]. Gallic acid was used as a reference standard for
15.73 µg of GAE/g FW. The results were superior in water extracts,
plotting calibration curve (Figure 1). Ten mg of gallic acid was
medium in ethanol and least in methanol in all the mango varieties
dissolved in 100 ml of 50% methanol (100 μg/ml) and then fur-
analysed except in the alphonsa mango. The order of TPC in mango
ther diluted to 6.25, 12.5, 25 or 50 μg/ml [8]. 1 ml of aliquots and
variants is given in following decreasing order: alphonsa mango>
standard gallic acid (10, 20, 30, 40 and 50 µg/ml) was positioned
nadusalai> senthuram> malgova> emmabasanth> kudhadath (Ta-
into the test tubes and 5 ml of distilled water and 0.5 ml of Folin
ble 1). TP extraction in water was significantly higher when com-
Ciocalteu’s reagent was mixed and shaken. After 5 minutes, 1.5
pared to the ethanolic and methanolic extracts (p≤0.05).
ml of 20% sodium carbonate was added and volume made up to
10 ml with distilled water. The reaction mixture was incubated at
Name of the Total Phenols (µg of GAE/ g of FW)
room temperature for 30 min with intermittent shaking for colour
Fruits Ethanol Methanol Water Total
development. The absorbance of the resulting blue colour was
measured at 765 nm using UV-VIS Double Beam Spectrophotom- Mango 539.23 ± 446.22 ± 889.72 ± 1875.17
(Banganapalli) 12.85 6.63 15.73
eter (Systronics, India). The linear equation of a standard curve
prepared with Gallic acid was used to determine the total phenolic Mango 176.37 ± 143.92 ± 193.31 ± 513.60
(Kudhadath) 8.46 4.82 8.70
contents of the extracts. The content of total phenolic (TP) com-
Mango 172.55 ± 147.27 ± 218.91 ± 538.73
pounds was expressed as µg/g Gallic acid equivalent (GAE) of fresh
(Emmabasanth) 3.79 10.59 7.17
weight.
Mango 273.26 ± 236.16 ± 301.67 ± 811.09
(Malgova) 4.70 0.34 4.43
Mango 322.21 ± 310.23 ± 357.29 ± 989.73
(Nadusalai) 16.47 16.46 14.33
Mango 265.77 ± 224.36 ± 327.51 ± 817.64
(Senthuram) 22.89 20.00 12.32
Mango 556.06 ± 493.94 ± 383.35 ± 1433.89
(Alphonsa) 4.89 8.01 1.94

Table 1: Total Phenols in three different extracts of Mango Pulp.

Discussion
Figure 1: Standard Curve of Gallic Acid –Total Phenol Assay.
Ribeiro., et al. [11] investigated the pulp composition of four
mango cultivars (Haden, Tommy Atkins and Ubá) at the ripening
The total phenolics content of the extracts were determined stage in relation to three components with antioxidant potential
colorimetrically, using the Folin-Ciocalteu method, as described by (total phenolics, carotenoids and ascorbic acid). Total phenolic
Singleton and Rossi [9], using gallic acid as a standard. Different al- compound content was estimated by the Folin-Ciocalteu reagent
iquots of sample solution were made up to 1ml each with distilled and total carotenoid content by spectrophotometry at 450 nm. The
water and added 1.5 ml of Folin– Ciocalteu reagent (previously di- contents of beta-carotene and total vitamin C (ascorbic acid and
luted 10 folds with distilled water), kept at room temperature for 5 dehydroascorbic acid) were quantified by high performance liquid
min and added 4 ml of 20% sodium carbonate (Na2CO3), and kept chromatography. Differences were found among the four mango
at room temperature for 30 min. The absorbance was read against cultivars in all the components analyzed. The content of phenolic
the blank at 750 nm. Here, blank as except test sample (extracts, compounds ranged from 48.40 (Haden) to 208.70 mg/100 g (Ubá);
standard). The total phenolics content was represented as milli- total carotenoid from 1.91 (Haden) to 2.63 mg/100 g (Palmer);
grams of Gallic acid equivalent per gram of dry mass (mg GAE/ g) beta-carotene from 661.27 (Palmer) to 2,220 microg/100 g (Ubá)
[10]. and total ascorbic acid ranged from 9.79 (Tommy Atkins) to 77.71
mg/100 g (Ubá). The results from several studies also show that
Results mangoes are a good source of antioxidants [8,12].
The preservation of vegetables and fruits phenolic content has
Eight genotypes of the mango fruit were evaluated for antioxi-
a great impact on the contribution of phenols not only in enzymat-
dant potential by several biochemical assays (DPPH, ABTS, ORAC,
ic browning reaction but also on nutrient content of the product as
FRAP, SASR and MCC) and tested for their polyphenol composition
antioxidants capacity. The results of this study indicate that all the
and vitamin C contents. The significance analysis demonstrated
fruits have phenolics that vary widely in different extracts and ag-
that the antioxidant capacity of Tainong 1 fruits were significantly

Citation: Jemima Beryl Mohankumar., et al. “Total Phenolic Content of Mango Pulp (Mangifera indica L.) Extracted using Three Different Solvents". Acta
Scientific Nutritional Health 3.9 (2019): 165-168.
Total Phenolic Content of Mango Pulp (Mangifera indica L.) Extracted using Three Different Solvents

168

higher than that of other genotypes, which was about 2.1–6.3-fold 5. Ma XW., et al. “Polyphenolic compounds and antioxidant
higher than Guifei assayed in ABTS, DPPH, ORAC and FRAP meth- properties in mango fruits”. Scientia Horticulturae 129
ods. Total polyphenols and flavonoids might be major contribu- (2011): 102-107.
tors to the antioxidant activity. Quantification of total phenolic
components would be used to evaluate antioxidant activities of 6. Xu BJ and Chang SK. “A comparative study on phenolic pro-
mango genotypes. The total polyphenols and flavonoids contents files and antioxidant activities of legumes as affected by ex-
showed a great variety amongst mango genotypes and high posi- traction solvents”. Journal of Food Science 72 (2007): S159-
tive correlation with the total antioxidant capacity. It is concluded 166.
that significant genotypic difference exists in the total antioxidant
7. Hayouni EA., et al. “The effects of solvents and extraction
capacity of mango fruits. Both total polyphenols and flavonoids
method on the phenolic contents and biological activities in
are major contributors to the total antioxidant capacity in mango
vitro of Tunisian Quercus coccifera L. and Juniperus phoeni-
fruit. They highlighted that antioxidant activity had significant dif-
cea L. fruit extracts”. Food Chemistry 105 (2007): 1126-1134.
ferences amongst the genotypes. The total polyphenols and flavo-
noids contents showed a similar variety [5]. 8. Lin JY and Tang CY. “Determination of total phenolic and fla-
vonoid contents in selected fruits and vegetables, as well as
Li., et al. [12], concluded that (a) there were diversities in anti-
their stimulatory effect on mouse splenocyte proliferation”.
oxidant contents and in vitro antioxidant capacities among culti-
Food Chemistry 101 (2007): 140-147.
vars and among groups, e.g., Guire No. 82 had superior polyphenol
content, flavonoid level, and in vitro antioxidant capacity, while the
9. Singleton VL and Rossi JA. “Colorimetry of total phenolics
highest level of α-tocopherol was observed in Renong No. 1; (b)
with phosphomolybdic-phosphotungstic acid reagents”.
higher contents of ascorbic acid, total polyphenol, total flavonoid,
American Journal of Ecology and Viticulture 16 (1965): 144-
and total in vitro antioxidant capability were observed in green
158.
peel mangoes than that in red peel or yellow peel mangoes; (c)
correlation analysis and principal component regression analy- 10. Roy N., et al. “A detailed study on antioxidant activity of stem
sis demonstrated that positive correlations exist among the mea- bark of Dalbergia sissoo Roxb. An Indian medicinal plant”.
sured antioxidants and total in vitro antioxidant capability, except Food Chemistry 126 (2011): 1115-1121.
α-tocopherol. Besides, polyphenol chemicals were deemed to be
one group of the most critical factors affecting the total in vitro 11. Machado Rocha Ribeiro Sônia., et al. “Antioxidant in Mango
antioxidant capability. Meanwhile, the strong synchronism of total (Mangifera indica L.) Pulp”. Plant Foods for Human Nutrition
polyphenol and flavonoid contents in pulp were found [12]. 62 (2007): 13-17.

12. Li L., et al. “Major Antioxidants and In Vitro Antioxidant

Conclusion
Capacity of Eleven Mango (Mangifera Indica L.) Cultivars”.
Total phenols which are a major contributor of antioxidant ac-
International Journal of Food Properties 17.8 (2014): 1872-
tivity was estimated in seven varieties of mango. Three solvents
1887.
were used for the extraction of TP. We obtained a greater extrac-
tion of TP in water. The commonly available variety of Alphonsa
mango had the highest TP. We suggest that other varieties popular Volume 3 Issue 9 September 2019
in the Northern parts of India may be analysed for the TP and AO © All rights are reserved by Jemima Beryl Mohankumar.,
content. et al.

Bibliography
1. https://mangifera.res.in/varieties.php

2. United States Department of Agriculture (USDA). USDA

national nutrient database for standard reference (2010).

3. Manthey JA and Perkins-Veazie Penelope. “Influences of Har-

vest Date and Location on the Levels of β-Carotene, Ascorbic
Acid, Total Phenols, the in Vitro Antioxidant Capacity, and
Phenolic Profiles of Five Commercial Varieties of Mango
(Mangifera indica L.)”. Journal of Agricultural and Food
Chemistry 57 (2009): 10825-10830.

4. Ribeiro SMR., et al. “Phenolic compounds and antioxidant

capacity of Brazilian mango (Mangifera indica L.) varieties”.
Food Chemistry 110 (2008): 620-626.

Citation: Jemima Beryl Mohankumar., et al. “Total Phenolic Content of Mango Pulp (Mangifera indica L.) Extracted using Three Different Solvents". Acta
Scientific Nutritional Health 3.9 (2019): 165-168.