Int J Adv Life Sci Res.

Volume 4(1) 22- 30

https://doi.org/10.31632/ijalsr.20.v04i01.004

International Journal of Advancement in Life Sciences Research

Online ISSN: 2581-4877
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Original Article

Eugenol Found by GC-MS Analysis of The Methanolic Extract of

The Fruit Pulp of Indigenous Mango (Mangifera indica L.) Variety,
Champa from Murshidabad District of West Bengal, India
1* 2 3 3
Mitu De , Susanta Ray , Subhasree Dutta , and Santi Ranjan Dey

1
Department of Botany, Gurudas College, Kolkata 700054, India.
2
Goalpara Tarun Sangha Junior High School, Khagra, Murshidabad, India.
3
Department of Zoology, Rammohan College, Kolkata, West Bengal, India.

*Correspondence E-mail : [email protected]

Abstract
Indigenous mango (Mangifera indica L.) varieties have unique aroma, taste, flavour, texture and size.
The interactions among volatile compounds and with sugars and acids have a role in the distinct
mango flavor. Murshidabad district of West Bengal has several indigenous mango varieties that have
unique attributes like taste and aroma. The indigenous Champa mango variety of Murshidabad district
has a distinct aroma very similar to that of the champaka flower (Michelia champaca L.). The present
investigation was to determine if there was any phyto-chemical which could be responsible for the
distinct and unique aroma present in the mango variety Champa. Eugenol, a phytogenic bioactive
component frequently found in diversified herbal plants was found by GC-MS analysis of the
methanolic extract of the fruit pulp of indigenous mango variety, Champa.

Keywords: Eugenol; Aroma; Champa; Indigenous mango variety; Murshidabad.

Introduction
Mango genetic diversity in India is very high distinct mango flavor. Sugars and acids
(Karihaloo et al. 2003). Ravishankar et al. enhance human perception of specific flavor
(2000) have reported that Mangifera indica L. notes in mango, including aromatics (Malundo
has extensive diversity due to alloploidy, et al, 2001). Aroma components in fruits also
outbreeding, continuous grafting and consist of aldehydes, alcohols, esters,
phenotypic differences arising from varied lactones, ketones, quinones and terpenes
agro climatic conditions in different mango (Costa et al, 2013). Both physical and
growing regions. Different varieties have chemical characteristics of mango viz. colour,
unique aroma, taste, flavour, texture and size. texture, flavor, sweetness are some of the
From the research work of Malundo and co qualities which determine consumer
workers it is believed that from the 150 volatile acceptability of the different mango varieties
compounds isolated from mango, it is probable (Mamiro et al, 2007). Aroma volatiles of
that only a few are critical to characteristic mango (Mangifera indica L.) determine
mango flavor (Malundo et al, 1996). The consumer acceptability and influence selection
interactions among volatile compounds and of parents for mango breeding programs
with sugars and acids have a role in the (Pandit et al, 2009).

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Int J Adv Life Sci Res. Volume 4(1)22-30

Mango (Mangifera indica L.) of the family crops under GI was very low in number than
Anacardiaceae is an economically important the crops actually eligible for registration. So,
tropical fruit globally. Mango is often referred there is an urgent need to aware about the
to as the ‘king of fruits.’ Mango is the national community/farmers about the importance of GI
fruit of India, Pakistan and Philippines. It is the
Volatile Organic Compounds (VOC) in
national tree of Bangladesh. Mango has high
mango
production, marketing and consumption rate
(Musharraf et al , 2016). Mango is cultivated in Jorge A. Pino and co workers at the Food
about 89 countries of the world (Yadav and Industry Research Institute, Havana have
Singh, 2017). worked extensively on the Volatile Organic
Compounds (VOCs) in various fruit like
Indigenous mango varieties of West Bengal
papaya (Pino et al, 2003), bullock’s heart fruit
Mango has rich intra-specific diversity . In (Pino et al, 2003), guava (Pino et al, 2002) and
India there are about one thousand indigenous mango ((Pino et al, 1989). As more research
or traditional varieties of Mangifera indica L. work was undertaken to study the aromatic
(Mukherjee, 1950). Malda and adjoining profiles of mango cultivars it was found that
district Murshidabad in West Bengal form one this group of compounds that determine the
of the largest mango yielding regions of the characteristic aroma of the fruit is commonly
country producing over 5 lakh tonnes. Mango present in small quantities, approximately 50
is the prime economic fruit in these two ppm or less. They are usually comprised of
districts. But after independence this mango mixtures of monoterpenes, sesquiterpenes,
germplasm came under threat due to rapid and volatile oxygenates (monoterpenes,
industrialization (Mukherjee, 1953). More than sesquiterpenes, esters, lactones, alcohols,
200 varieties of mango were known from aldehydes, ketones, volatile fatty acids, some
Malda and Murshidabad districts only as degradation product of phenols, and some
mentioned but extensive survey in 2011-2014 carotenoids) (Bender et al., 2000; Pino et al.,
showed that only 53 varieties are now 2005; Lebrun et al., 2008; Pandit et al.,
available (Pal et al, 2017). 2010; Li et al., 2017).
However even after massive genetic erosion Several research groups have worked on the
there still exists a rich collection of indigenous volatile components in different mango
mango varieties in Murshidabad district (Dutta cultivars in several countries viz. Australia
et al, 2020). There are many indigenous (Bartley, 1988), Brazil (Andrade et al, 2000),
varieties are rarely found outside Murshidabad Cuba (Pino et al, 1989). In 2005 Pino’s
districts viz. Anaras, Begum Pasand, Champa, research group carried out studies to
Kohitoor, Kahinoor, Mulayam Jaam, determine the Volatile organic compounds
Ranipasand, Saranga. Generally, old (VOCs) profile of different mango cultivars.
traditional mango cultivars are maintained The research work led to the identification of
either in home or village gardens, personal more than 300 compounds including esters,
orchards, within the precincts of temples and aldehydes, ketones, aliphatic alcohols, γ and
on public sites such as schools. This system δ-lactones, and mono- and sesquiterpenes
continues due to the mango’s religious and (Pino et al, 2005). Researchers are of the
cultural importance. Custodian farmers have opinion that the volatile components of mango
been maintaining, promoting and adapting a vary between different mango varieties and
number of indigenous mango varieties on their consequently the aroma profile also differs
farms. These custodian farmers protect, (MacLeod et al, 1984).
nurture and propagate the rich mango legacy
Aroma chemistry
of India (De, 2015).
Aroma chemistry is complex. The
Most of the fruit crops growing in India had
fragrance/aroma/smell of any flower is
special characters which are distinct from the
never really the consequence of a single
fruits growing in other countries. These special
chemical compound. Flowers give off a
characters facilitates the registration of fruit
complex mix of volatile organic chemicals, and
crops under GI but the number of registered

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Int J Adv Life Sci Res. Volume 4(1)22-30

whilst not all of these contribute to the aroma,

a significant number impact the fragrance to
varying degrees. So often we can’t point
to single chemical compounds as being the
cause of flowers’ scents, we can identify those
compounds that have a major impact on the
aroma that our noses detect. In 2007 Jorge A.
Pino’s research group found aroma volatile
constituents of Colombian varieties of mango
(Quijano et al, 2007).
Unique scented indigenous Mango
varieties of Murshidabad district
The indigenous mango variety Anaras has a
distinct aroma of pineapple. If a person is Fig. 1. Indigenous Mango variety Champa,
blindfolded and asked to taste the fruit there is collected from Ajimgange, Murshidabad
a high probability that the mango variety Brief description of the fruit of indigenous
Anaras could be mistaken for a pineapple. In mango variety, Champa from Murshidabad
face the literal Bengali translation of ‘Anaras’ is district, West Bengal:
pineapple. The famous but very rare mango
variety Kohitur is said to have a unique taste Mature fruit is short in length (9 cm), with a
and aroma. The Champa mango variety has a narrow width, ratio of length/width is small,
distinct aroma very similar to that of the shape of cross section is circular, color of the
champaka flower (Michelia champaca L.). The skin is green and yellow, and density of
word Champa is the Bengali word for the lenticels is dense. The color contrast between
scented flower champaka. Michelia champaca lenticels and skin is strong, with large size of
L. belongs to the family Magnoliaceae the lenticels. The corkiness is due to the
presence of the lenticels is present, shallow
Objective of the study cavity at stalk is present. Shallow neck is
The objective of this study was to determine if present. Shape of ventral shoulder is rounded
there was any volatile substance which could outwards, shape of dorsal shoulder is rounded
be responsible for the distinct and unique downwards. Groove in the ventral shoulder is
aroma present in the mango variety Champa. absent, and bulging on the ventral shoulder is
Also we wanted to see if there was any bio- also absent. Shallow sinus is present; depth of
active compound which was similar to any sinus is low. Bulging at the proximal stylar scar
compound reported in Michelia champaca L. is absent, point at the stylar scar is medium.
using the with Gas Chromatography-Mass Diameter of the stalk attachment is small.
Spectrometry analysis method. Predominant color of skin of the ripe fruit is
yellow green, speckling of skin is weak,
Materials and Methods thickness of skin is thin, adherence of the flesh
to the skin is medium. Main color of the flesh
Collection of samples of the ripe fruit is medium yellow. Firmness of
The fruit of indigenous mango Champa variety the flesh of the ripe fruit is soft, with medium
was collected from the orchard of a ‘custodian juiciness, and medium texture of flesh (Fig. 1,
farmer’ in Murshidabad. 2 & 3).

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Int J Adv Life Sci Res. Volume 4(1)22-30

was used. The column (HP5) was fused silica

50 m × 0.25 mm I.D. Analysis conditions were
20 minutes at 100°C, 3 minutes at 235°C for
column temperature, 240°C for injector
temperature, helium was the carrier gas and
split ratio was 5:4. The sample (1 μl) was
evaporated in a split less injector at 300°C.
Run time was 22 minutes. The components
were identified by GC coupled with MS.
Identification of components
Interpretation of the mass spectrum, GC-MS
was conducted using the database of National
Institute Standard and Technology (NIST)
having more than 62,000 patterns. The
spectrum of the unknown components was
compared with the spectrum of the known
Fig. 2. Fruit of the indigenous mango variety, components stored in the NIST library. The
Champa from Murshidabad district of West name, molecular weight and structure of the
Bengal components of the test materials were
ascertained.

Results
The identification of phytochemical
compounds was based on the peak area,
retention time and molecular formula. The
compound was identified as eugenol (Table 1).
Eugenol, a phytogenic bioactive component is
frequently found in diversified herbal plants
possessing well-defined functional attributes
(Fig.4 & 5). Eugenol is mainly found in clove,
cinnamon, nutmeg, tulsi and pepper. Eugenol
is the major volatile constituent of clove bud
essential oil. Eugenol is declared as GRAS
(generally recognized as safe) by World
Fig. 3. Fruit of the indigenous mango variety,
Health Organization (WHO) and is considered
Champa (close up view).
as non-mutagenic (Khalil et al, 2017). Methyl
Fruit pulp collection eugenol is directly derived from eugenol, a
product from phenylalanine (an essential
Fruit pulp from the mango variety Champa
amino acid) through caffeic acid and ferulic
was collected from Murshidabad district. The
acid via the shikimate pathway (Herrmann and
mango fruit pulp was stored in the refrigerator
Weaver, 2009). It is a common
for further use. Methanolic extract were
phenylpropanoid found in many plant species,
prepared for the analysis.
particularly in spices and medicinal plants (Tan
GC-MS and Nishida, 2012). Methyl eugenol has been
The experiment was carried out in the Central reported to be present in Michelia champaca L
Instrumentation Facility, Bose Institute, (Sanimah et al, 2008) and is postulated to be a
Kolkata, India. For GC-MS Thermofisher GC compound impacting the sweet fragrance of
Michelia champaca L.

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Int J Adv Life Sci Res. Volume 4(1)22-30

Figure 4. GC MS spectra and NIST result for identification of compound, eugenol.

Table 1. The compound identified as eugenol. Following details were observed from
https://pubchem.ncbi.nlm.nih.gov/compound:

Compound IUPAC Name Molecular Molecular Synonyms

Formula Weight
Eugenol 2-methoxy-4- C10H12O2 164.2 g/mol 4-Allyl-2-methoxyphenol,
prop-2- Allylguaiacol,
enylphenol 1,4-Eugenol;
1-Hydroxy-2-methoxy-4-
propenylbenzene,
2-Methoxy-4-allylphenol,
Eugenolum,
5-Allylguaiacol,
Eugenic acid

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Int J Adv Life Sci Res. Volume 4(1)22-30

varieties most are of seedling origin and are

found to be growing as heirloom varieties from
generation to generation (Dinesh et al, 2015).
Some of these unique mango cultivars are
popular and also registered under
Geographical Indication of Goods
(Registration and protection) Act 1999. Names
of the indigenous mango varieties registered
under geographical indication (GI) include
Appemidi Mango of Western Ghats,
Karnataka, Laxman Bhog Mango of Malda
district, West Bengal, Khirsapati (Himsagar)
Fig. 5. Structure of Eugenol.
Mango of Malda district, West Bengal and
Discussion Fazli Mango grown Malda in West Bengal,
Mango Malihabadi Dusseheri of Lucknow
Aroma is a major factor that influences the district, Uttar Pradesh, and Gir Kesar Mango
quality and consumer acceptance of mango of Gujarat state (Rani & Kishore, 2013).
products (Liu et al, 2020). Detailed study of the
aromatic components could improve the Apart from the famous indigenous mango
understanding and also facilitate the control of varieties which have already got a GI tag there
critical quality parameters that could influence are several unique traditional mango varieties
mango processing. Eugenol is a natural in several pockets. Their uniqueness could be
compound that has several pharmacological attributed to certain phyto-chemicals. But in
activities, action on the redox status, and recent years farmers are abandoning the
applications in the food and pharmaceutical indigenous mango varieties in favour of more
industry. Eugenol (4-allyl-2-methoxyphenol) is lucrative commercial mango cultivars. Farmers
a phenolic compound from the class of are keen to maintain commercial mango
phenylpropanoids and the main component of cultivars for fresh consumption and income
clove. Often eugenol has been used in the generation. Also the original habitats of local
food industry as a preservative, mainly due to mango have been rapidly changing in
its antioxidant property (Zhang, et al. 2009). response to biotic, economic and other
pressures in recent years (Subedi et al. 2004).
Methyleugenol is used as a flavouring agent in
jellies, baked goods, non-alcoholic beverages. So on one hand there is an urgent need to
It has been used as a fragrance ingredient in identify unique mango varieties and novel
perfumes, toiletries and detergent. It is phyto-chemicals which account for the unique
possible the Eugenol or its derivative in the aroma and other characteristics. Detailed and
indigenous mango variety Champa is well documented information about the
responsible for the unique flavor. available genetic material together with a
Genetic diversity analysis of mango broad, well maintained varietal diversity are
germplasm is not uniform throughout all the essential for future mango breeding efforts.
states of India viz. Bajpai et al, 2016; Dash This should also include local varieties (Subedi
et al., 2005), which may have a low market,
and Hota, 1997. However in India, the mango
gene pool has not been explored fully. Each of but high breeding value. Identifying indigenous
the traditional varieties of mango has a unique mango varieties with unique features and
taste and flavor (Mehta, 2017). In India many marketing them could lead to higher profit
mango varieties grow under specific climate. among the mango orchard owners. This could
This specific type of climate is responsible for be a deciding factor for conserving a
acquiring some unique characteristics for that indigenous mango variety and its legacy for
particular indigenous mango variety. So many posterity. Brand development of mango is
indigenous mango varieties which have unique needed in such a way that it would lead to
taste, texture or aroma and they are found integrated growth of that crop (Banerjee,
restricted to certain areas. Amongst these 2011). These indigenous varieties will survive

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Int J Adv Life Sci Res. Volume 4(1)22-30

alongside modern varieties if they are resources. Once lost, they are lost forever. It
characterised by distinctive traits that make needs to be cherished and preserved
them relevant in the farming system or otherwise there is a great risk of losing the
demanded in the market (Berg, 2009). genetic resource.
Conclusion Acknowledgments:
In West Bengal, the mango gene pool has not This work has been carried out with the
been explored fully. The scientific information financial assistance of a Research Project of
relating to aromatic constituents as well as the West Bengal Biodiversity Board (WBBB) to
sensory characteristics of indigenous mango the first author as principal investigator. The
cultivars of West Bengal is limited. There are authors would like to thank Prof Sujoy
several unique varieties of mango, which need Dasgupta of Bose Institute and Mr. Smriti
to be identified and if possible given a Kumar Majhi, Central Instrument Facility, Bose
geographical identity. Further detailed Institute, Kolkata 54 for their support and
investigations are needed to understand the assistance to carry on the GC-MS analysis.
cause of their uniqueness. Efforts are also The authors thank the Principal, Gurudas
needed to locate the places linked with a College, Kolkata 700054 and Principal,
particular variety of mango and thereafter Rammohan College, Kolkata 700019 for
attempt for registration of geographical identity providing support to carry on this work.
of such local varieties. The genetic diversity
embedded within the indigenous mango Conflicts of Interest:
varieties are a unique genetic resource. Plant
The authors declare that there are no conflicts
genetic resources are among the most
of interest regarding the publication of this
vulnerable of all non-renewable natural
work.
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