Mangifera sylvatica (Wild Mango): A new cocoa butter alternative

Abstract

Cocoa butter is the pure butter extracted from cocoa beans and is a major ingredient in the chocolate industry. Global production of cocoa is in decline due to crop failure, diseases and ageing plantations, leading to price fluctuations and the necessity for the industry to find high quality cocoa butter alternatives. This study explored the potential of a wild mango (Mangifera sylvatica), an underutilised fruit in south-east Asia, as a new Cocoa Butter Alternative (CBA). Analyses showed that wild mango butter has a light coloured fat with a similar fatty acid profile (palmitic, stearic and oleic acid) and triglyceride profile (POP, SOS and POS) to cocoa butter. Thermal and physical properties are also similar to cocoa butter. Additionally, wild mango butter comprises 65% SOS (1, 3-distearoyl-2-oleoyl-glycerol) which indicates potential to become a Cocoa Butter Improver (an enhancement of CBA). It is concluded that these attractive properties of wild mango could be prompted by a coalition of policy makers, foresters, food industries and horticulturists to promote more widespread cultivation of this wild fruit species to realise the market opportunity.

Full text

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Mangifera sylvatica (Wild Mango):
A new cocoa butter alternative
Sayma Akhter1, Morag A. McDonald1 & Ray Marriott2
Cocoa butter is the pure butter extracted from cocoa beans and is a major ingredient in the chocolate
industry. Global production of cocoa is in decline due to crop failure, diseases and ageing plantations,
leading to price uctuations and the necessity for the industry to nd high quality cocoa butter
alternatives. This study explored the potential of a wild mango (Mangifera sylvatica), an underutilised
fruit in south-east Asia, as a new Cocoa Butter Alternative (CBA). Analyses showed that wild mango
butter has a light coloured fat with a similar fatty acid prole (palmitic, stearic and oleic acid) and
triglyceride prole (POP, SOS and POS) to cocoa butter. Thermal and physical properties are also similar
to cocoa butter. Additionally, wild mango butter comprises 65% SOS (1, 3-distearoyl-2-oleoyl-glycerol)
which indicates potential to become a Cocoa Butter Improver (an enhancement of CBA). It is concluded
that these attractive properties of wild mango could be prompted by a coalition of policy makers,
foresters, food industries and horticulturists to promote more widespread cultivation of this wild fruit
species to realise the market opportunity.
Cocoa butter (CB) is a light yellow fat obtained from beans of the cocoa plant (eobroma cacao L.). It is one of
the unique natural fats highly demanded by food, pharmaceuticals and cosmetic industries1,2. Cocoa butter is
the major ingredient of the chocolate industry3. Palmitic acid (C16:0), stearic acid (C18:0), oleic acid (C18:1)
and linoleic acid (C18:2) account for more than 98% of the total fatty acids4 in cocoa butter. is is the only
commercially available natural fat which is rich in saturated and monounsaturated fatty acids,13.6–15.5% of
1,3-dipalmitoyl-2-oleoyl-glycerol (POP), 33.7–40.5% of 1-palmitoyl-3- stearoyl-2-oleoyl-glycerol (POS) and
23.8–31.2% of 1,3-distearoyl-2-oleoyl-glycerol (SOS)5,6. ese relatively simple triglycerides in cocoa butter con-
fer its desirable melting proles prized by the confectionery industry, being solid at 20 °C and melting between
27–35 °C which is appreciated by consumers as well as desirable in confectionery applications7. Moreover, the
price of cocoa butter is one of the highest among all tropical fats and oils5,7,8. According to ICCO (2015), the price
of cocoa butter more than doubled between 2005 and 20159, from $1433/tonne to$3360/tonne (Supplementary
Fig. 1). Cocoa is cultivated on a land area of over 70,000 km2 worldwide10 while Africa (68%), Asia (17%) and
America (15%) contribute the major proportion of global production of CB10. According to ICCO9, annual global
cocoa production was reported to be more than 4 million tonnes per season. However, global demand for cocoa is
growing annually by 2 to 3% due to low productivity, price uctuations and uncertainty in supply (Supplementary
Fig. 1), which has forced the confectionery industry to seek CBAs2,5 from other natural sources11. Cocoa butter
equivalents (CBEs) are commercially available fats containing a similar mixture of triacylglycerol to cocoa butter
that can be mixed with cocoa butter up to 5%12. Very few tropical fats are considered to beCBAs but include those
sourced from illipe butter, kokum butter, shea butter and mango (Mangifera indica L.) butter13. e mango kernel
contains about 7–15% fat that is rich in palmitic, stearic and oleic acids11,13. Cocoa butter from the domesticated
mango species, M. indica, is a natural fat containing high saturated and monounsaturated fatty acids containing
symmetrical triglycerides such as POS (10 to 16%), SOS (25 to 59%) and POP (1 to 8.9%)13. ese are relatively
simple triglyceride combinations which are desirable for confectionery applications, especially in chocolate pro-
cessing. M. indica kernels have therefore been heavily researched for their potential as a cocoa butter alternative14.
Wild mango (Mangifera sylvatica Roxb.) belongs to the Anacardiaceae family. It is found in Bangladesh, India,
China, Cambodia, Myanmar, Nepal and ailand (Fig.1)15. It is one of the genetically closest species to M. indica
in the world16 but it underutilized and unmarketed in its native provenance Bangladesh as well as in other tropical
countries due to a lack of information and awareness of it’s potential value as a source of food, nutrition or medi-
cine. Perhaps counterintuitively given its underutilisation, the species is already threatened in Bangladesh17 due to
habitat loss and deforestation, but is not aorded any conservation protection due to its lack of documented value.
1School of Environment, Natural Resources and Geography, Bangor University, Gwynedd, LL57 2UW, UK.
2Biocomposites Centre, Bangor University, Gwynedd, LL57 2UW, UK. Correspondence and requests for materials
should be addressed to S.A. (email: sayma_sust@yahoo.com)
received: 27 April 2016
Accepted: 25 July 2016
Published: 24 August 2016
OPEN

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e seed germination rate and early growth of seedlings indicates that this species could be easily domesticated
and incorporated into small-scale forestry programs15. ere is growing evidence of benecial medicinal proper-
ties, such as a recent study showing that M. sylvatica leaves possess thrombolytic properties that could lyse blood
clots18. e leaves can also be used as antidiarrheal drugs19. Until now, no research has been conducted on its
market potential which will ultimately promote domestication and commercialisation of the species. erefore,
the present study constitutes an assessment of the potential of M. sylvatica as a CBA for the food, pharmaceutical
and cosmetic industries. In this study, the fatty acid and triglyceride composition, and the physiochemical and
thermal properties of M. sylvatica were determined and compared to those of the domesticated mango, and cocoa
butter to assess the potential for M. sylvatica as a new source of cocoa butter.
Results
Fatty acid Prole and Triglyceride compositions. Wild mango butter (WMB) is a light yellow fat that
is not greasy to touch and has a characteristic nutty avour. WMB is a rich source of saturated fatty acids (Fig.2a).
WMB consist of three major fatty acids, namely palmitic acid (C16:0), stearic acid (C18:0) and oleic acid (C18:1)
(Fig.2b). e saturated fatty acid content of M. sylvatica (56%) approximates that of DMB (57%) but is lower
compared to CB (65%). Stearic acid, oleic acid and palmitic acid account for 95% total fatty acid in WMB from
M. Sylvatica followed by CB (96–97%) and DMB (94%). Apart from that, M. sylvatica butter contains small
amounts of arachidic acid and linoleic (also known as Omega- 3 Fatty Acid), which is similar to CB and DMB
(Fig.2b). Triglycerides are complex mixtures of a variety of fatty acids, which are the major constituents in fats
and oils. e major triglycerides found in WMB are 1, 3-distearoyl-2-oleoyl-glycerol (SOS), 1-palmitoyl-2-oleoyl-
3-stearoyl-glycerol (POS) and 1, 3-dipalmitoyl-2-oleoyl-glycerol (POP) which is also the main features of cocoa
butter (Table1). POP, POS and SOS account for 79% for WMB and 82–85% for CB (Fig.3h). is similarity in
fatty acid and triglyceride proles indicates considerable potential for WMB to be used as a source of cocoa butter
alternative.
Physical and thermal properties of WMB. e saponication value, glycerol percentage, iodine value,
free fatty acid percentage, moisture content, specic gravity and refractive index were determined for the wild
mango butters as important parameters of butter quality. In WMB the saponication value is slightly lower than
CB (Fig.3a) which means WMB consists of long chain carbon molecules but is close to DMB. WMB contains
bigger carbon chain molecules so it has fewer glycerol molecules as indicated by glycerol percentage (Fig.3b). e
iodine value of WMB is slightly higher than CB (Fig.3d) but close to DMB. An elevated iodine value and acid
value indicates high susceptibility of fat to oxidative rancidity due to the high degree of unsaturation. Moisture
content and free fatty acid content in WMB was high compared to other butter samples (Fig.3c,g) which also
indicate the possibility of WMB oxidation. ese results suggest that proper and controlled processing can pro-
duce high quality butter with decreased degradation. e refractive index and Specic gravity of WMB butter was
Figure 1. Global and Local Distribution of Wild Mango (Mangifera sylvatica). Spatial position of the site
locations were plotted in global geo-political boundary available from Esri (http://www.arcgis.com/) and species
presence locations were plotted in administrative map of Bangladesh using ArcGIS (version 10.3) soware.

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very similar to CB and DMB (Fig.3f,g). is indicates the double bond present in WMB is similar like CB and the
weight of WMB is very similar to CB. WMB has a melting point close to CB though DMB has a higher melting
point (Fig.4). WMB is characterised by one leading peak around 16.18 °C with a “shoulder” around −4.71 °C.
M. sylvatica is similar to CB (Table2) where the main melting peak appeared around 20 °C. A complete melting
of WMB was observed around 26 °C and 27 °C for CB. e results from M. sylvatica are very dierent from
M. indica where the main melting peak was observed around 16.88 °C but with multiple shoulders and with a very
high melting point observed around 53 °C (Fig.4).
Discussion
e majority of studies report palmitic acid, stearic acid, oleic acid and linoleic acid to be the major fatty acid
components of CB20. Minor components of lauric acid (C 12:0), myristic acid (C 14:0), linolenic (C 18:3) and ara-
chidic acid (C 20:0) have also been reported20. e main dierence between CB and WMB observed in this study
was in the palmitic acid content, 27% and 6% respectively (Supplementary Table 2). Other studies of triglyceride
content of DMB have reported highly variable results; POP (6–16%), SOS (2–59%) and POS (1–74%) and POO,
SOO, SOA, OOO7,13 compared to CB which has more consistent concentrations of POS (37–47%), SOS (26–33%)
and POP (16–23%) (13, 20). TGAs in WMB are similar to CB, where POP, POS and SOS are the major TGAs but
with a higher percentage of SOS (65%). WMB contains a slightly lower SFA content compared to CB but the fatty
acid prole is comparable (Fig.2a,b). erefore, it is evident that the fatty acid and triglyceride composition of
WMB is close to that of CB derived from eobroma cacao, indicating good prospects for WMB to be a source of
cocoa butter alternative.
Key parameters in conferring high fat quality, distinctive avour and aroma in butters are the saponication
and acid values. WMB has a lower saponication value than CB (2, 21) which means the fatty acids in WMB are
signicantly longer carbon chain compounds (Fig.5a). Such long chain fatty acids (saturated and unsaturated) are
prone to oxidation and breakdown which provides characteristic avours and aromas High acid values indicate
breakdown of triglycerides into free fatty acids (FFA) relating to inadequate processing and storage conditions.
Cocoa butter is reported to have acid values in the range of 0.42 to 3.11%21. e acid value of fat extracted from
DMB varies from 1.22 to 7.487,22. Our analyses showed that WMB has a signicantly higher acid value compared
to CB which suggests there might be processing or storage problems. With respect to iodine values, the higher the
value the more reactive, less stable, soer the fat and hence more susceptible to oxidation and rancidication7. In
general, the iodine value for CB was found to be 34–38 g I2/100 g8,23,24 and for DMB 40–75 g I2/100 g7,22. e iodine
Figure 2. (a) Total saturated and unsaturated fatty acid content; Fig.1. (b) Fatty acid prole of in WMB, DMB
and CB (CBD and CBND).
Triglycerides Triglycerides WMB DMB CBD CBND
1,3-dipalmitoyl-2-oleoylglycerol POP √ √ √ √
1-palmitoyl-2-oleoyl-3-stearoyl-glycerol POS √ √ √ √
1,3-distearoyl-2-oleoyl-glycerol SOS √ √ √ √
Trioleoyl-glycerol OOO √ √
1-Arachidoyl-2-Oleoyl-3 Linoleyolglycerol AOLo √ √
1,2 -Palmitoyl-3-Linoleoylglycerol PPLo √ √
1-stearoyl-2,3-dioleoyl-glycerol SOO √
1-stearoyl-2-oleoyl-3-Arachidoyl-glycerol SOA √
Table 1. Triglyceride Prole of M. sylvatica butter, M. indica butter and Cocoa Butter.

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value of WMB fat is higher than that of CB. is study, again suggesting that adequate storage will be essential if
it is to be used more widely. e moisture content of WMB is higher than CB which may render more susceptible
to microbial attack and oxidation. However, the moisture content is easily managed during the extraction process.
On the positive side, there is a growing body of evidence that higher moisture content butters produce more low
fat chocolate which may help to prevent obesity, heart diseases, diabetics, stroke and arthritis23. Indeed, there is
on-going research to produce low fat chocolate by adding water into the CB24. Manipulation of the extraction
Figure 3. Physical properties of butter samples (a) saponication value (b) glycerol (c) free fatty acid (d) iodine
value (e) moisture content; (f) refractive index; (g) specic gravity (h) major triglyceride percentage.
Figure 4. Melting prole of dierent butter using DSC.

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process to best manage moisture levels will eliminate the need to add water to the nal product. e refractive
index and specic gravity of WMB was very similar to CB. It has been suggested that the physicochemical char-
acteristics of WMB can be manipulated through controlled processing, chemical or physical rening and natural
blending processes to adjust the properties of WMB to CB14,25. e melting point is important to determine the
storage temperature. e melting temperature of CB is slightly higher than WMB which could be due to of the
higher saturated fatty acid content (Fig.2a) as previously noted23. Similar results have been reported for CB by
many researchers2,14,26–28. So, there were some signicant similarities in the physical and thermal properties of
WMB compared to CB which again shows the potential of WMB to be used as a cocoa butter alternative.
Chocolate commands an enviable position among food products due to its premium cost, taste and unique
physicochemical properties20. e consumption of chocolate products has signicantly increased worldwide29
whilst 30% of the world’s cocoa crops have been destroyed by pests and disease and are deteriorating due to
climate change and ageing plantations. Demand is increasing and supply is inadequate as cocoa is cultivated
in only a few tropical countries, making its availability unstable, expensive and subject to price uctuations20,
(Supplementary Fig. 1). Moreover, poor quality harvests and some technological problems such as fat blooms and
high tempering times during chocolate production make it necessary for the food industries to look for alterna-
tives to CB and intensive eorts are ongoing to nd suitable cocoa butter alternatives29. Cocoa butter alternatives
are divided into three subgroups (Supplementary Fig. 2). Cocoa butter replacers (CBRs) are non-lauric fats with a
fatty acid prole similar to cocoa butter, but a completely dierent triglyceride composition (e.g. PEE, SEE) Cocoa
butter substitutes (CBSs) are lauric plant fats (containing lauric acid), chemically totally dierent to cocoa butter
(e.g. major TGAs LLL, LLM, LMM), with some physical similarities; suitable only to substitute cocoa butter to
100% and oen incompatible with CB30). Cocoa butter equivalents (CBEs) are non-lauric plant fats, which are rel-
atively similar in their physical and chemical properties (e.g. major TGAs are POP, POS and SOS) to cocoa butter
and can be proportionately mixed without aecting the properties of the cocoa butter. CBEs can be either cocoa
butter extenders (CBEXs) which is a subgroup of CBEs not mixable in every ratio with cocoa butter or Cocoa
butter improvers (CBIs) which have a higher solid triglyceride (SOS) content; used for improving so cocoa
butters20,31. Chocolate and confectionery industries give priority to fats which are rich in palmitic acid or stearic
acid and are based on symmetric (POP-rich and SOS-rich) fats. Some research has shown that SOS rich fat con-
fers a higher solid fat content which inhibits fat blooms and decreases the tempering time26. erefore, SOS-rich
fat could be used as a suitable raw material for the production of temperature-resistant hard butters in tropical
countries2 and could also be used to improve the quality of so cocoa butter2. Generally, lauric acid and hydro-
genated fats are used to replace CB; these increase the levels of cholesterol whereas CBEs contain high oleic and
stearic acids, which do not alter the levels of cholesterol in blood. us, CBEs represent a healthier and promising
alternative to CB. CBEs used up to now are tropical SOS rich fat butters from species such as Shea (Vitellaria par-
adoxa), Kokum (Garcinia indica), Illipe (Shorea stenoptera), Mango (Mangifera indica) and Sal (Shorea robusta)
butter and usually blended with palm (Elaeis guineensis) kernel oil stearin rich in POP31. Palm kernel oil consists
of higher amounts of lauric acid, and relatively lower stearic and oleic acid than cocoa butter. e producing of
CBA from palm oil needs intensive processing8. Palm kernel oil is used in preparing CBA as it is a very rich source
of POP (51%). Sal butter is green in colour which limits its use in chocolate and confectionery products13. It is
feasible to lighten butters but it is a very energy intensive procedure and costly, so industries prefer light coloured
butters11. Kokum butter is grey coloured and mainly used as a CBE by blending with Mahua (Madhuca longifolia)
and Phulwara (Madhuca butyracea) butter32. However, the extraction is only practiced at cottage scale and has no
industrial application as yet32,33. Shea butter is known to have the highest unsaponiable fat content (up to 10%)
of any natural fat and the highest iodine value 52–56 (g Iodine/100 g fat) and is used as a cocoa butter substitute34
in the European chocolate and confectionery industry35. Illipe and mango butters can be used directly as CBE and
Mango Butter (Mangifera indica) is comparatively good quality CBEs although the melting point (34–43 °C) is
quite high20,31. ere is therefore not enough reliable source of CBE available from natural fat sources20. Our study
suggests that WMB is a potential high quality new CBE or improver as the fatty acid and triglyceride composition
are very similar to CB as are the physical parameters.
However, going beyond an industrial utility, wild fruit is an important source of food, medicine and income
for forest dwellers, tribal and marginalized rural people36. ere are many wild fruits available in the forests
that are underexploited. Moreover, information on their nutritional value and economic potential are unknown.
Adding value to underutilized products through processing for products that have market value could generate
a way to conserve those species and help to generate alternative income sources and reduce household pov-
erty37. Additionally, collection and processing of these products can reduce household vulnerability to shocks
and seasonal variations in other income sources38. For example, shea kernels from Vitellaria paradoxa are widely
exported for use in the international cosmetic and chocolate industries. e annual value of total exports of
shea kernels from Africa was estimated at USD 30 million in 200439 and they represent one of Burkina Faso’s
main export commodities40. Moreover, income from shea kernels has been shown to contribute as much as 12%
of total household income for poor households and 7% of total household income for better-o households40.
Sample Tonset(°C) Tos et(°C) Tpeak(°C) Enthalpy change (J/g)
Mangifera sylvatica butter − 7.20 25.88 16.18 16.3383
Mangifera indica butter − 5.36 58.00 16.66 10.4353
Cocoa Butter Deodorized 14.64 26.99 20.27 42.1099
Cocoa Butter Non-Deodorized 14.58 26.99 20.34 45.7108
Table 2. Melting Characteristics of dierent butter samples.

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In Bangladesh, there are 47 edible wild forest fruits available41, an important one of which is the wild mango
species (M. sylvatica). Wild mango is a multipurpose tree species used for multiple purposes, including edible
fruits, pickles, fodder, fuelwood, vegetable, plywood, tea chest and match boxes41,42. A close genetic relationship
between M. indica and M. sylvatica has been reported43,44 which indicates that M. sylvatica may have the potential
to full nutritional and livelihood needs. It is underutilized in Bangladesh as well as in other tropical countries
due to a lack of awareness of it’s potential as a source of food and no established market demand45. However, this
research conrms that this underutilized wild mango has the potential to be used as a unique source of cocoa
butter alternative.
Bangladesh is one of the most densely populated countries in the world, with 2.14 million hectares for-
est area46. Millions of the poor and forest dwellers earn their livelihood from the forest47. erefore, there is a
socio-economic imperative to allow access for these forest dependent people to the natural resource. However,
nding alternative income generating activities can secure income, improve livelihoods and conserve forest
resources sustainably48. ere is enormous potential for the development of a wild mango kernel based enterprise
in Bangladesh as well as in other tropical countries for the production of wild mango butter. is will not only
provide raw materials for the chocolate and confectionery industries but also oer opportunities to empower
forest dependent people and small-scale farmers. ere is therefore an urgent need to promote the domesti-
cation and commercial plantation of wild mango species to satisfy global and local demand for Cocoa Butter
Alternatives. A recent study shows that it can be domesticated and introduced in small-scale forestry programs15.
However, larger scale plantings will require eld trials and an improved knowledge of the species silviculture. e
current study may lead to the beginning of a domestication and commercialization of this wild under-utilized
fruit species. However, more research on chocolate production using this butter and silviculture of this species is
necessary to fully capture the value of this wild mango species. Additionally, collaboration between foresters, hor-
ticulturists, the food industry and policy makers is required to promote the domestication and commercialization
of M. sylvatica fruits of Bangladesh and other tropical countries.
Figure 5. Collection, processing and preparation of wild mango butter from M. sylvatica (a) Mangifera sylvatica
tree (b) fruit of Mangifera sylvatica with big kernel in le and fruit of Mangifera indica with big pulp in right (c) seed
of Mangifera sylvatica (d) kernel of Mangifera sylvatica (e) Mangifera sylvatica seeds aer retting (f) chopping the
seeds (g) sun drying the kernels (h) cocoa butter (i) butter obtained from Mangifera sylvatica (Photo Credit goes to
Sayma Akhter).

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Material and Methods
Sample Collection, preparation, extraction and analysis. Mature fruits of M. sylvatica were col-
lected from Cox’s Bazar, Bangladesh (Fig.1), during April to June, 2014. Aer collection, fresh fruits were retted,
de-pulped and sun-dried. e nuts were then separated from the kernel by using a hand betel nut cutter (Fig.5).
Phosphine fumigation was carried out before nuts were sent to Bangor University, UK for further processing.
Mango butter was extracted using the SC-CO2 (Supercritical Carbon Dioxide Fluid Extraction) method. We
obtained two cocoa butter samples from Callebaut chocolate industry (UK) and purchased 99% pure Mangifera
indica butter (Domesticated Mango Butter, DMB) from the Soapery. Finally, analysed the physical (saponication
value, iodine value, moisture content, specic gravity, refractive index, acid value, and glycerol percent), chemical
(fatty acid prole and triglyceride composition) and thermal (melting prole) parameters of Mangifera sylvatica
butter (Wild Mango Butter, WMB) and compared the results of WMB with the three other butter samples.
Wild mango Butter extraction by SC-CO2. e wild mango kernels were extracted using Supercritical
Fluid Extraction method2. A total 18.85 kg of dry ground mango kernel samples were loaded into the extraction
vessel. e continuous methods of SC-CO2 extraction were carried out at pressures of 50 MPa, temperatures
of 40 °C and at constant CO2 ow rate of 30 kg/hour. When pressurization initiated, the CO2 from the cylinder
passed through the chiller at 0 °C and was pumped into the extraction vessel by a high-pressure pump. e fat
was extracted from fat-rich CO2 by separators at one end of the instrument. Two separators were used though the
entire process with the rst separator being at xed pressure and temperature of 80 MPa and 40° respectively. e
second separator was maintained at room temperature and 55 MPa and desiccated the samples. CO2 was recir-
culated throughout the run time. Yield was calculated on a dry weight basis at the end of the process as g fat/kg
mango kernel.
Fatty acid and triglycerides proling. e fatty acid composition of mango (M. indica and M. sylvatica)
and cocoa butter (Deodorized and non-Deodorised) samples were done by GC (PerkinElmer Clarus 680)-MS
(PerkinElmer Clarus 600 C). Five to seven mg of frozen butter dissolved in 1 ml of heptane and 0.05 ml of 1 N
Methanolic NaOH were shaken at room temperature for two minutes. Aer 2 minutes when the two layers were
separated the lower layer was discarded and the supernatant) used for GC-MS analysis. e analysis was done in
triplicate. On the other hand, a direct infusion mass spectrometry method (API 150 EX MS System) used for the
determination of Triglycerides. e nebulizer gas was N2. Scanning done for mass 100 to 1000 in an ESI Positive
mode with a ow rate of 90 μ l/min. Samples were run once for triglyceride proling2,7 and percentage of triglyc-
erides were calculated based on the peak intensity.
Physio-chemical and thermal properties. Determination of saponication value, glycerol percentage,
acid value, iodine value, moisture content, refractive index and specic gravity were carried out according to
methods describes by2,7,14,49. Quantitative analyses were performed in triplicate and the results expressed as aver-
age ± standard deviation. One-way ANOVA was conducted to see any signicance dierence among four types
of butter.Dierential scanning calorimetry (DSC) used to monitor the melting proles of the samples. A modied
method of Yamoneka29 was used for this analysis. e method followed was heating- cooling - heating cycle. 1st
cycle (− 20 °C to 60 °C) and 2nd cycle (60 °C to −20 °C) was done to erase thermal memory and also to get rid of
any unwanted materials. e nal cycle (− 20 °C to 60 °C) was recorded to get the melting prole of the samples.
e heating rate was 10 °C/min and cooling rate was 2 °C/in.
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Acknowledgements
European Union funded this research through the FONASO (Forest and Nature for Society) Erasmus Mundus
Joint Doctoral Programme. Dr. Gee-Sian Leung and Dr. Luis Martin (BioComposite Centre, Bangor University,
UK) are thanked for their cooperation during the butter preparation and sample analysis. Special thanks to Helen
Simpson (Senior Laboratory Technician, SENRGy) and Sarah Chesworth (Laboratory Technician, ECW) from
Bangor University, UK for their assistance during the laboratory work. Special thanks to Md. Basir Al Mamun
(Assistant Conservator of Forests in Bangladesh Forest Department) and Professor Dr. Mohammed Jashimuddin

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Scientific RepoRts | 6:32050 | DOI: 10.1038/srep32050
from Institute of Forestry and Environmental Science, Chittagong University, Bangladesh. We also would like to
thank the forest department ocials, eld assistants and local people for their invaluable help in collecting and
processing fruits.
Author Contributions
S.A., M.M. and R.M. planned the research. S.A. collected sample, laboratory experiments, analysed data and
prepared manuscript dras. R.M. assisted with laboratory experiments. M.M. and R.M. reviewed and edited the
manuscripts.
Additional Information
Supplementary information accompanies this paper at http://www.nature.com/srep
Competing nancial interests: e authors declare no competing nancial interests.
How to cite this article: Akhter, S. et al. Mangifera sylvatica (Wild Mango): A new Cocoa butter alternative.
Sci. Rep. 6, 32050; doi: 10.1038/srep32050 (2016).
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