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Effect of vegetable oil in the solubility of capsaicinoids extracted from capsicum Chinense Bhut Jolokia

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Abstract

Capsicum chinense Bhut Jolokia is in fact the world's hottest known chilli pepper owing to the presence of alkaloids collectively known as capsaicinoids. It has become important to establish efficient technique for extraction of capsaicin and its analogues. The present research was conducted to analyze the effect of vegetable oils in the solubility of capsaicinoids. The capsaicinoids extracted in the oils were quantified by UV spectrophotometer and Phosphomolybdic acid reduction methods. Total phenol content in the extracts was estimated by Folin-Ciocalteu method. Among the tested oils gingelly oil, Olive oil and Sunflower oil showed high capsaicin content. The result of this investigation clearly concludes that the different oils used for the extraction showed diverse solubility of capsaicinoid. Sunflower oil extract can be formulated as topical applications. Neem and castor oil extract can be used as pesticide. Oils would be better solvent for the efficient extraction of capsaicinoids for its biological applications.impact 0.7
Research Article
EFFECT OF VEGETABLE OIL IN THE SOLUBILITY OF CAPSAICINOIDS EXTRACTED FROM
CAPSICUM CHINENSE BHUT JOLOKIA
NAGOTH JOSEPH AMRUTHRAJ, J.P. PREETAM RAJ, L. ANTOINE LEBEL*
Department of Plant Biology and Biotechnology, Loyola College, Chennai 600 034, Tamilnadu, India. Email: antonest12@gmail.com
Received: 5 December 2013, Revised and Accepted: 28 January 2013
ABSTRACT
Capsicum chinense Bhut Jolokia is in fact the world’s hottest known chilli pepper owing to the presence of alkaloids collectively known as
capsaicinoids. It has become important to establish efficient technique for extraction of capsaicin and its analogues. The present research was
conducted to analyze the effect of vegetable oils in the solubility of capsaicinoids. The capsaicinoids extracted in the oils were quantified by UV
spectrophotometer and Phosphomolybdic acid reduction methods. Total phenol content in the extracts was estimated by Folin- Ciocalteu method.
Among the tested oils gingelly oil, Olive oil and Sunflower oil showed high capsaicin content. The result of this investigation clearly concludes that
the different oils used for the extraction showed diverse solubility of capsaicinoid. Sunflower oil extract can be formulated as topical applications.
Neem and castor oil extract can be used as pesticide. Oils would be better solvent for the efficient extraction of capsaicinoids for its biological
applications.
Keywords: Vegetable oils, Capsicum, Bhut Jolokia, capsaicin, capsaicinoids, phosphomolybdic acid,
INTRODUCTION
The genus Capsicum belongs to the Solanaceae family which
includes more than 30 species [1]. Capsicum chinense Naga Jolokia
(also called Bhut Jolokia, Naga Morich, Bih Jolokia etc.) is in fact the
world’s hottest known chili pepper with a pungent level of 879,953
to 1,001,304 SHU [2]. The Naga chilli has massive scope in domestic
as well as international market due to its major high capsaicin
content. Capsicum fruits are known for its flavor, color and to add
tang and taste to the insipid food. It is cooked as whole or ground
and alone or in combination with other flavoring agents, primarily in
the curry powder and pickles. Despites of its indispensable spice
capability, this genus has recognizable pharmacological properties
due to the presence of alkaloids collectively known as capsaicinoids
[3]. The economic, nutritious and pharmacological significance of it
is responsible for its high demand. In addition to food additive in our
daily diet, the medical applications of capsaicin make this compound
very popular. It is currently available as various topical
pharmaceutical forms (ointments, high-dose dermal patches,
creams, large bandages) for a number of diverse clinical conditions,
such as the pain relief of peripheral neuropathy, symptomatic
treatment of arthritis, muscle and joint pains and other types of
neurogenic pains such as Herpes zoster. It is worn in the treatment
of obesity and ulcer in the forms of oral herbal supplements. Its
chemical agent used in riot control and personal defense pepper
spray. Furthermore, it was demonstrated that capsaicin induces
apoptosis in various cancer cells [3,4]. Because of the increasing
demand by consumers for hot and spicy foods and also the
increasing use in medicine and pharmacy, it has become important
to establish a sensitive, accurate and simple technique for extraction
of capsaicin and its analogues. The use of essential oil in herbal
medicine would be base for the development of novel potent drugs.
[5]. Hence the present study was conducted to find the solubility of
capsaicinoids in different vegetable oils extracted from Capsicum
chinense Bhut Jolokia fruits.
MATERIALS AND METHODS
Sample Collection
Capsicum chinense fruits were obtained from Manipur, North India.
The morphology of the fruit shape, color, seed color and size of the C.
chinense were examined according to the description of [6]. The
fruits were dried by traditional method i.e. sun dried for a day,
ground, sieved through 2030 mesh and kept in air tight containers
until further process. The standard Capsaicin (8-methyl-N-vanillyl-
trans-6-nonenamide) was purchased from Sigma Chemical Co, St.
Louis, MO, USA. Chemicals used for the analysis were from Merck.
Extraction of capsaicinoids by cooking oils and medicinal oils
500mg of sieved chilli powder was boiled at 65oC for an hour with
5ml of following oils: Brassica juncea (mustard oil), Helianthus
annuus (sunflower oil), Olea europaea (olive oil), Cocos nucifera
(coconut oil), Elaeis guineensis (palm oil), Ricinus communis (castor
oil), Azadirachta indica (neem oil), Arachis hypogaea (ground nut
oil) and Sesamum indicum (gingelly oil). The extract was centrifuged
for 10,000 rpm for 10 min. The supernatant was subjected to
quantification of capsaicinoids.
Quantification by UV spectrophotometer
The simple linear regression curve was plotted using standard
capsaicin purchased from Sigma Chemical. A stock solution of one
milligram capsaicin per milliliter of ethanol was dissolved and
different concentrations were prepared from the stock solution 10µg
to 100 µg. The optical density was recorded at 280nm. The linear
regression equation was generated using the online Statistics and
forecasting software (www. wessa.net). The capsaicinoid extracted
from the different oils were estimated by UV visible
spectrophotometer (Hitachi- U1800). The optical density was
recorded at 280nm. The capsaicin concentrations in samples were
calculated using capsaicin linear regression equation and it was
expressed as mg/g of the chili powder and finally converted to
Scoville Heat Unit.
Quantification of Total capsaicin
Capsaicin is a protoalkaloid which is responsible for the pungency of
chilli, the quality of the chilli fruit; extract of oleoresin is determined
by the capsaicin content. The phenolic group in capsaicin reduces
the phosphomolybdic acid to lower acids of molybdenum; the
resulting compound is blue in color and was read at 650 nm the
color intensity directly proportional to the concentration of
capsaicin [7]. Standard capsaicin solution was diluted to range of
100µg, 80 µg, 60 µg, 40 µg and 20 µg and liner regression curve was
generated using the online Statistics and forecasting software
(www.wessa.net). To 0.1 ml of extracted oil into a glass stoppered
test tube and added 10 ml of dry acetone (added about 25gm
Vol 7, Suppl 1, 2014 ISSN - 0974-2441
Lebel et al.
Asian J Pharm Clin Res, Vol 7, Suppl 1, 2014, 48-51
49
anhydrous sodium sulphate into 500ml acetone of analytical grade
at least 1 day before use) and kept it in shaker for an hour. The
content was centrifuged at 10000 rpm for 10 min. 1ml of the clear
supernatant was pipette out and evaporated to dryness in a hot
water bath. The residue was dissolved in 5ml of 0.4% sodium
hydroxide solution and 3ml of Phosphomolybdic acid were added
and were kept for an hour. The solution was centrifuged to remove
the floating debris. The colored solution was directly read at 650 nm.
Capsaicin content calculated from the standard curve was expressed
as mg/g of the chilli powder and finally converted to Scoville Heat
Unit.
Quantification of Total Phenol
Total phenol contents of hot peppers were analyzed using the
modified Folin- Ciocalteu reagent method [8]. To 0.1ml of extract
1ml of 1/10 dilution Folin-Ciocalteu reagent (Fisher) and 2ml of
7.5% (w/v) Na2CO3 was added. After vortexing for 10 sec, the
mixture was incubated at 45ºC in a water bath for 15 min. Samples
were allowed to cool at room temperature before reading the
absorbance at 765 nm using Hitachi UV-Vis spectrophotometer. A
blank was prepared from without the extract. The standard curve
was was generated using the online Statistics and forecasting
software (www. wessa.net).
Scoville Heat Unit (SHU) Conversion
According to the commonly accepted Scoville organoleptic test, the
spicy strength of the investigated samples was calculated by
converting the capsaicin content expressed in grams of capsaicin per
gram of pepper. This conversion to Scoville heat units was done by
multiplying the capsaicin content in pepper dry weight by the
coefficient corresponding to the heat value for pure capsaicin, which
is 1.6 × 107 [9].
RESULTS
The simple linear regression curve was plotted for standard
capsaicin purchased from Sigma Chemical. The linear regression
equation was generated using the online Statistics and forecasting
software (www. wessa.net). The capsaicin contents obtained in
μg/ml were converted to Scoville heat units in order to classify them
according to their various pungency levels. The total capsaicin
content in gingelly oil, olive oil and sunflower oil showed high
pungency level with 1,379,357 SHU, 1,148,175SHU and
879,596.3SHU respectively(Table 1 and Fig 1). The UV estimation
showed range of 150 000 to 300 000 SHU in tested oils, but castor
oil showed 62,884 SHU which is said to be less pungent (Table1 and
Fig 2). The total phenol estimated in the extracted oils showed
contemporise the amount of total capsaicin estimated (Fig 3). Hence
the current study summaries that the different oils used for the
extraction showed diverse solubility of capsaicinoid.
DISCUSSION
The present study was focused on the solubility of capsaicinoids in
vegetable oils categorized into cooking and medicinal oils. The
pungency or hotness of chilli peppers is determined by the
concentration of capsaicinoids produced as a secondary metabolite
in the fruit and it is often presented as “Scoville Heat Units SHU [10].
The capsaicinoids content in the extracts were estimated
spectrophotometrically (Anan et al., 1996). The UV-VIS
spectrophotometric method is one of the most inexpensive and
accessible for capsaicin quantification. However analysis is
restricted to capsaicin solutions with microgram-level
concentrations [11]. The choice of suitable solvent for the extraction
of capsaicioind is crucial, to consider the solubility of pigments
which can impact subsequent purification steps. Capsaicinoid yields
were significantly affected by time and duration. The rate of
capsaicin solubility was relatively high for the solvents, temperature
and duration. The optimum extraction conditions reported for air
dried C. frutescens was 120 C for 15 min with acetone (Barbero et
al., 2008). Temperature lower than the 700C recommended in the
AOAC [13] official method of analysis for fruits and vegetables was
followed to minimize loss of capsaicinoids through volatilization. In
the present study, for the better yield of capsaicinoids from Bhut
Jolakia the extraction was carried out at 65C for one hour. The
presence of water during extraction impacts the hydrophilic
properties of solvents and interactions with the capsaicinoid
compounds, resulting in varying solubility of capsaicin [12, 14].
Previous research has proved that cosmetic-grade plant products
can be extracted using hexane, ethanol, or vegetable oil. Capsaicin is
fairly resistant to acids and alkali solutions at room temperatures
[15]. It is only slightly soluble in water [16], but more soluble in
ethanol and vegetable oils [17]. The vegetable oils used in this
experiment include cooking and medicinal oils. Vegetable oils
penetrate deeply into the skin than paraffin oil [18]. The chemical
compounds of olive oil contribute to its overall therapeutic
characteristics [19]. Rheumatoid arthritis patients receiving olive oil
benefits pain reduction and reduced morning stiffness were
reported [20]. Topical application of olive oil caused a significant
reduction in stratum corneum integrity and induced mild
erythemain. Sunflower seed oil preserved stratum corneum
integrity, with improved hydration and without erythema, [21].
Capsaicin extracted in the oil would enhanced the penetration of an
anti-inflammatory agent through human skin as well as it will
reduced reduce skin irritation. Plant oils protects the skin by
dissolving the sebum secreted from oil glands, thus support
evaporation from the skin [22]. So we suggest that capsaicin
extracted in sunflower oil can be used in the topical application and
olive oil can be used in cooking food for better tang and hotness.
Capsaicin as agrochemicals has been proven as pesticides against
cotton crop pest [23]. Extracts of capsicum fruits has been used as
an repellent to some species of stored product beetles such as
Sitophilus zeamais Motschulsky and Tribolium castaneum [24].
Moreover plant oils, are safe, low in cost and eco friendly, beside
their effectiveness against several pests [25]. Hence the neem oil and
castor oil used in the extraction of capsaicinoids can b e formulated
and tested against the pests.
CONCLUSION
Capsicum species are used worldwide not only as a food, but also as
medicine for its astounding therapeutic applications such as
anticarcinogenis, antioxidants, suppression of fat accumulation, anti-
inflammatory, antimicrobial and used in topical creams. Hence this
study concludes that the vegetable oils would be better solvent for
the efficient extraction of capsaicinoids for its biological
applications. In order to get more tang and hot olive oil and giglley
oil can be used for cooking in the food. For the ointments and cream
preparation Sunflower oil can be used. The capsaicinoids in the oils
can be emulsified and it can be used as nano carriers. Future studies
are still needed to learn more about oils when used in combination
with capsaicin as part of topical cream and pesticides.
ACKNOWLEDGEMENT
My grateful thanks to Rev.Sr. Mercy Margret .B. ICM and Mrs.
Sargunam amma for their support and encouragement.
Tabel.1 SHU units of Capsaicnoids content in oils extracts analysed by UV-Visible spectrophotometer and reduction of Phosphomolybdic
acid
Oil used
Scoville Heat Unit (SHU)
UV Method
Pungency level
Phosphomolybdic
Pungency level
acid
Cooking oil
280,464
***
633,116
***
234,047
***
879,596
***
244,491
***
1,148,175
***
298,451
***
865,997
***
295,549
***
483,527
***
Lebel et al.
Asian J Pharm Clin Res, Vol 7, Suppl 1, 2014, 48-51
50
275,242
***
623,766
***
169,949
***
1,379,357
***
Medicinal oil
62,884
**
590,619
***
69,503
**
135,054
***
*** Very highly pungent (> 80,000SHU), ** Highly pungent (25,000 70, 000SHU)
M usta rd O il
S u n fl o w e r O i l
Oliv e O il
C o co n ut O il
P a lm Oil
G roundnut O il
G in g el ly O il
C a sto r O il
N e em O il
0
5
10
15
20
V eg eta bl e O ils
capsaicin in µ g/m l
U V M et ho d
Fig.1: Shows the amount of capsaicin in different vegetable oils
used in extraction- quantified by UV spectrophotometer
M usta rd O il
S u n fl o w e r O il
O li ve O il
C o co n ut O il
P a lm Oil
G ro undnut O il
G in ge ll y O il
Casto r O il
Ne em O il
0
20
40
60
80
100
Ph osp ho m oly b di c a c id r ed uc tio n m e th od
V eg eta ble O ils
capsaic in in µg/m l
Fig.2: Shows the amount of capsaicin in different vegetable oils
used in extraction- quantified by Phosphomolybdic acid
reduction method
M ustard O il
S u n fl o w e r O i l
Oliv e O il
C o co n ut O il
P a lm O il
G roundnut O il
G in g e lly Oil
C a sto r O il
N e em O il
0
50
100
150
F oli n- C ioc al teu m e tho d
V eg eta bl e O ils
capsaic in i n µ g/ ml
Fig.3: Shows the Total phenol content in different vegetable oils
used in extraction- quantified by Folin Ciocalteu method
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... The maceration of dehydrated Capsicum spp. in vegetable oils has been recently carried out using some comestibles (olive, corn, sunflower, safflower, coconut, and palm) and medicinal oils (mustard, neem, ricinums, ground nut, and ginger) at different temperatures and extraction times and then filtered and centrifuged to separate the solids to obtain the non-aqueous extracts of chili (Guadarrama-Lezama et al. 2012;Amruthraj 2014). The extraction of components from C. chinense (Bhut Jolokia) in different vegetable oils depends on the solubility of capsaicin and other compounds present in the oil, and it was also an efficient solvent to extract capsaicinoids for biological applications (Amruthraj 2014). ...
... The maceration of dehydrated Capsicum spp. in vegetable oils has been recently carried out using some comestibles (olive, corn, sunflower, safflower, coconut, and palm) and medicinal oils (mustard, neem, ricinums, ground nut, and ginger) at different temperatures and extraction times and then filtered and centrifuged to separate the solids to obtain the non-aqueous extracts of chili (Guadarrama-Lezama et al. 2012;Amruthraj 2014). The extraction of components from C. chinense (Bhut Jolokia) in different vegetable oils depends on the solubility of capsaicin and other compounds present in the oil, and it was also an efficient solvent to extract capsaicinoids for biological applications (Amruthraj 2014). ...
... The extracted profile obtained depends on the solvent polarity and other physico-chemical properties of the particular solvent (see Table 5) and from the extraction conditions (time and temperature) (see Table 6). Therefore, less polar solvents as hexane, acetone, and methanol can extract carotenoids easily (Arimboor et al. 2015) and are less recommended to extract capasaicinoids than polar aprotic solvents such as acetone and acetonitrile and even for polar protic solvents such as methanol and ethanol (Amruthraj 2014). ...
... Only a few works have been performed using comestible oils and medicinal oils at different temperatures and extraction times. For example, Amruthaj et al. [74] extracted capsaicin from habanero chili using mustard, sunflower, olive, coconut, palm, groundnut, gingelly, castor and neem oils as extractants, finding that olive and gingelly oils were the best extractants for capsaicin due to the presence of other phytochemicals that could enhance capsaicinoids properties. ...
... [107] Capsaicinoids are easy soluble in fat, so vegetable oils could be employed as extractants with higher yields. [6,74] Solute and solvent ratio This is a critical factor that influences the mass transfer in the capsaicinoids extraction process, because a larger volume of solvents helps to accelerate the diffusion process. For conventional techniques, the tendency is to reduce the ratio of mass and volume of solvent, and in many instances this increases the extraction volume obtained. ...
... Water is generally added to the solvent such as methanol or ethanol to enhance the extraction of capsaicinoids, because the presence of water during extraction impacts the hydrophilic properties of solvents and interactions with the capsaicinoids compounds, resulting in varying solubility of capsaicin. [74] When analytes are extracted at low concentrations, the rate of extraction is not affected by the analyte concentration, but rather by the rate of mass transfer; therefore, the chemical properties of solvent should be chosen to ensure solvation and the release of the analyte. [108] Extraction temperature This is a fundamental parameter important for capsaicinoids extraction, which depends on the matrix, extraction technique and process conditions. ...
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Different techniques have been employed to extract capsaicinoids from habanero chili pepper (HCP). Conventional extraction techniques use long extraction times and environmentally unfriendly solvents, while other techniques are limited by the polarity of the capsaicinoids, which cause their underestimation. Recent advanced extraction techniques of capsaicinoids have been developed to reduce time, increase yields and preserve their pungency degree and bioactive properties. This review aims to highlight the trends in extraction techniques used on capsaicinoids recovery from habanero chili peppers, providing a critical and detailed discussion of the main parameters influencing the extraction yield and analyzing data to establish the most viable and efficient extraction technique.
... The maceration of dehydrated Capsicum spp. in vegetable oils has been recently carried out using some comestibles (olive, corn, sunflower, safflower, coconut, and palm) and medicinal oils (mustard, neem, ricinums, ground nut, and ginger) at different temperatures and extraction times and then filtered and centrifuged to separate the solids to obtain the non-aqueous extracts of chili (Guadarrama-Lezama et al. 2012;Amruthraj 2014). The extraction of components from C. chinense (Bhut Jolokia) in different vegetable oils depends on the solubility of capsaicin and other compounds present in the oil, and it was also an efficient solvent to extract capsaicinoids for biological applications (Amruthraj 2014). ...
... The maceration of dehydrated Capsicum spp. in vegetable oils has been recently carried out using some comestibles (olive, corn, sunflower, safflower, coconut, and palm) and medicinal oils (mustard, neem, ricinums, ground nut, and ginger) at different temperatures and extraction times and then filtered and centrifuged to separate the solids to obtain the non-aqueous extracts of chili (Guadarrama-Lezama et al. 2012;Amruthraj 2014). The extraction of components from C. chinense (Bhut Jolokia) in different vegetable oils depends on the solubility of capsaicin and other compounds present in the oil, and it was also an efficient solvent to extract capsaicinoids for biological applications (Amruthraj 2014). ...
... The extracted profile obtained depends on the solvent polarity and other physico-chemical properties of the particular solvent (see Table 5) and from the extraction conditions (time and temperature) (see Table 6). Therefore, less polar solvents as hexane, acetone, and methanol can extract carotenoids easily (Arimboor et al. 2015) and are less recommended to extract capasaicinoids than polar aprotic solvents such as acetone and acetonitrile and even for polar protic solvents such as methanol and ethanol (Amruthraj 2014). ...
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BhutJolokia, chilli pepper has health-benefiting effects for which it is used in foods and pharmaceutical products. It is highly perishable at higher moisture content, and sun-drying of it requires more time in comparison to other modern methods. This article presents the oven drying of BhutJolokia at 40°C and 70°C, for the determination of its drying characteristics. The drying conditions, physicochemical properties and the models were studied to describe the drying behaviour. The moisture content reduced significantly (p<0.05) after drying. The sun-drying (ambient, 30ºC) and oven drying at 40ºC and 70ºC, took nearly about 27 h, 20.5 h and 13.5 h, respectively, to reduce the final moisture content in each of the samples to 9% (approximately). The half-time (t 1/2) for drying of chilli under the sun, at 40ºC
... Tere is signifcant research demonstrating that capsaicinoid compounds, derived naturally from chilli, have antitumor properties, both in vitro and in vivo (Šaponjac, et al. 2014;Amruthraj, et al. 2014). ...
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Oleoresin is a mixture of volatile and nonvolatile components available in whole extract of natural herb or spice. It principally comprises essential oils and resin. Lemongrass oleoresins come from the Cymbopogon species, which grow in the tropical and subtropical regions of the world. Oleoresin of lemongrass is a dark green-colored viscous liquid having a characteristic lemon aroma and flavor and is mostly used as a flavoring ingredient. The lemon prefix in the lemongrass specifies the characteristic lemon-like odor, which is due to the availability of citral content (mixture of two isomeric aldehydes, geranial and neral). It has been utilized in synthesizing flavors, perfumes, cosmetics, detergents, and in the food and pharmaceutical industries. Different methods are used to extract the lemongrass essential oil, but steam distillation is the most suitable method as it doesn’t alter the quality of the obtained oil. The chemical composition of lemongrass oil varies depending on its extraction methods, genetic differences, harvest period, photoperiod, plant age, farming practices, and geographical origin. Lemongrass essential oil has shown several biological activities, including antimicrobial, antifungal, antiprotozoan, antioxidant, antidiarrheal, antimutagenic, antiinflammatory, antimalarial, antinociceptive, antihepatotoxic activities, etc. Lemongrass oil is a potent food preservative because of its extraordinary antifungal and antibacterial activities.
... Chili pepper is also suitable for the diets of the obese and is useful in the control of cancer of the stomach and colon (TayebRezvani et al., 2013;Dang et al., 2014). Chili pepper fruits are low in sodium and free cholesterol (Chigoziri and Ekefan, 2013) and are used in sauces, soups, stews and generally as a flavouring agent (Reyes-Escogido et al., 2011;Amruthraj et al., 2014). The total antioxidants are completed by phenolic compounds, which occur in peppers in connection with sugars (Materska et al., 2003a, b). ...
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... Chili pepper is also suitable for the diets of the obese and is useful in the control of cancer of the stomach and colon (TayebRezvani et al., 2013;Dang et al., 2014). Chili pepper fruits are low in sodium andfree cholesterol (Chigoziri and Ekefan, 2013) and are used in sauces, soups, stews and generally as a flavoring agent (Reyes-Escogido et al., 2011;Amruthraj et al., 2014). ...
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