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Shamsun Naher et al / Int. J. Pharm. Phytopharmacol. Res. 2015; 5 (1): 35-40
35
Research Article Article ID: 402
Studies on physico-chemical properties, GC-Mass and ED-XRF analysis of fatty oil of Capsicum annum
Linn (dry chili) in Bangladesh
Shamsun Naher*
1
, Tanvir ahmed
1
, Md. Abu Salam
1
, S. M. Mahmudul Hassan
2
,
Shahin Aziz
2
and Mala Khan
2
1
Department of Chemistry, Jagannath University, Dhaka 1100, Bangladesh
2
BCSIR Laboratories Dhaka, Bangladesh Council of Scientific and Industrial Research,
Dr. Qudrat-E-Khuda Road, Dhaka-1205.
*Corresponding Author: Dr. Shamsun Naher, Associate Professor, Department of Chemistry, Jagannath University, Dhaka1100, Bangladesh.
Email: shamsunnaher2002@yahoo.com Tel: 088-01720250388
Abstract
Capsicum Annum linn. is locally known as dry chili in Bangladesh. It is used as spices all over the world. In this
study fatty oil of the two varieties of dry chilly from two different places of Bangladesh were analyzed by GC-
MS. Dry chili (Capsicum Annum linn.) Fatty oil has been widely used1 industrially as an ingredient of many
products, e.g. hot sauces, canned fish, ginger beer, as well as for some pharmaceutical products. Fatty oil of
two varieties of dry chili was analysis by GC-MS and their composition were established. The GC-MS analysis
showed that, Dinajpur’s dry chili contained total nine fatty oils. These were Tetradecanoic acid,methyl ester
(2.72%); 9-Hexadecanoic acid ,methyl ester (1.55%); Hexadecanoic acid ,methyl ester (17.40%); 9,12-
Octadecadienoic acid(Z,Z)- methyl ester (44.64%); 10-Octadecanoic acid ,methyl ester (24.33%); 9-
octadecenoic acid(z)-methyl ester (2.39%); Octadecanoic acid ,methyl ester (3.76%); Octadecanoic
acid,9,10,12-trimethoxy-,methyl ester (1.43%); 4-tert-butyl-2-methoxyquinoline- 3-phenyl-2-piperidino-2-propen-
1-a 4-butyl-2,2-dimethyl-2H-1-benzopyran (1.80%). On the other hand, Chittagong’s dry chili contained total
ten fatty oils. These were Dodecanoic acid,methyl ester (1.99%); Tetradecanoic acid,methyl ester(5.69%); 9-
Hexadecanoic acid ,methyl ester (1.14%); Hexadecanoic acid ,methyl ester (30.15%); 9,12-Octadecadienoic
acid(Z,Z)- methyl ester (29.30%); 9-octadecenoic acid(z)-methyl ester(17.82%); 8-octadecenoic acid,methyl
ester(3.08%); Octadecanoic acid ,methyl ester (5.48%); Octadecanoic acid,9,10,12-trimethoxy-,methyl ester
(2.02%); Methoxylate ester of methyl oleate (3.34%). Elemental analysis was also performed by ED-XRF
Analyzer and was observed the mineral contents in fatty oil of the samples. In the minerals, % of K was major in
both the samples (85.16% for Dinajpur and 82.54% for Chittagong).
ISSN (Online) 2249-6084 (Print) 2250-1029
International Journal of Pharmaceutical and
Phytopharmacological Research (eIJPPR)
[Impact Factor – 0.852]
Journal Homepage: www.eijppr.com
Article info
Article History:
Received 11 August 2015
Accepted 24 August 2015
Keywords:
Capsicum Annum Linn., Gas
chromatography and mass
spectrometry (GC-MS), ED-XRF
Analyzer, physicochemical properties,
fatty oil.
Shamsun Naher et al / Int. J. Pharm. Phytopharmacol. Res. 2015; 5 (1): 35-40
36
1. INTRODUCTION
Capsicum annuum Linn. is an important part of human diet since 7500 BC
2
. Chili peppers originated in America
3
. Chilies holds all the best
properties for which it is consider as a food. It has been used as a food for many of years. Chilies are high in vitamin C (about twice that of
citrus fruits) Even after cooking it only loses 30 percent of its vitamin C. Dried chilies are very high in vitamin A Red chilies are a great source of
β-carotene
4-8
. Chilies have antibacterial qualities, and contain bioflavonoid, anti-oxidants most common in apple juice. It is effective in
protecting against cancer. Capsaicin cream is used to lower the sensation of pain in such conditions as arthritis, and other painful chronic
condition
9-11
. Pungency in chili is due to the alkaloid “capsaicin” contained in the pericarp and placenta of fruits, it produces mild to intense spice
when eaten. Numerous reviews have summarized the conditions under which capsaicin-containing pharmaceutical agents can best be used
12-
14
. Chili is considered as one of the most important commercial spice crops and is widely used universal spice, named as wonder spice.
Different varieties are cultivated for varied uses like vegetable, pickles, spice and condiments. Chili (botanically known as Capsicum annuum L.;
Capsicum frutescene L.), also called red pepper belongs to the genus capsicum, under the solanaceae family. Chilies are referred to as chillies,
chile, hot peppers, bell peppers, red peppers, pod peppers, cayenne peppers, paprika, pimento, and capsicum in different parts of the world.
Capsicum Annum (chili) plant oils & extracts have been used for a wide variety of purposes of many thousand of years. Chili grows best at 20–
30°C. Growth and yields suffer when temperatures exceed 30°C or drops below 15°C for extended periods. The crop can be grown over a wide
range of altitudes from sea level up to nearly 2100 meter
15-16
. However many researches have been carried out on dry chili (Capsicum annuum
Linn.), but no systematic research on comparative studies has been reported on the fatty oil of dry chili in Bangladesh. Some disagreement
about the presence of its constituents was observed. Therefore, present work was undertaken to carry out a complete investigation of the fatty
oil of nutmeg of two varieties including its physico-chemical properties along with GC analysis.
2. MATERIALS AND METHODS
The fresh Bangladeshi dry chili is available in the local markets. The dry chili was collected from two different parts of Bangladesh (Dinajpur and
Chittagong). The collected samples were washed clearly by water to remove dust materials. Then they were dried. Finally the dried chili was
ground by Fritsch mortar grinder, Germany for one hour. Then the powder was sieved prior to the extraction process. The mean particle
diameters obtained were 0.25 and 0.50mm.
2.1 Physico-chemical studies
17
Physico-chemical characteristics of seed such iodine value (pet-ether extract), moisture, ash, color, Solubility, crude fiber, protein,
carbohydrate, food energy, specific gravity, refractive index were determined by following the standard procedures
18-19
, and the results were
shown in Table-1.
2.1.1 Isolation of fatty acids and preparation of methyl ester:
The neutral lipids-were extracted from the air-dry powdered fruit successively with light petroleum ether (40-60
0
c), n-hexane and chloroform in a
Soxhlet apparatus each for 22h, respectively. Pet-ether, hexane and chloroform extracts were mixed to equal weight (2mg) to analyze the fatty
acid in the fruit. The sample was kept in a nitrogen atmosphere in a refrigerator. Fruit sample was analyzed according to the method reported
by griffin
20
for esterification and fatty acid methyl ester (FAME) extraction.
2.1.2 Preparation of Standard fatty acid methyl ester (FAME)
Eleven standard free fatty acids (Caprilic acid ; Lauric acid; myristic acid ; palmitic acid; palmitoleic acid; linoleic acid , oleic acid; stearic;
arachiaonic acid; behenic acid; and lignoceric acid) were individually weighed . About 5mg of each was taken in a reaction tube and
BF
3
CH
3
OH reagent (5ml) was added to it. The mixture was boiled for 5 min. Hexane (5 ml) was added to it and boiled for further 1 min. After
cooling the tube a solution of saturated salt was added and vortexes. Then the upper layer containing methyl esters was transferred to a vial
with anhydrous sodium sulfate at the bottom. Then the ester was filtered through syringe filter and transferred to a small vial (2ml). The solvent
was concentrated by blowing nitrogen gas and stored in a refrigerator before analysis by GC.
2.2 GC-MS Analysis
The essential oil of Capsicum annuum Linn. (Dry chili) of two varieties were analyzed by Electron Impact Ionization (EI) method on GC-17A gas
chromatograph, coupled to a GC-MS 2010 plus mass spectrometer; fused silica capillary column temperature of 40˚ C ( was held 2 min) was
maintained with carrier gas helium at a constant pressure of 90kPa. Samples were injected by splitting with the split ratio 10. Essential oil
sample was dissolved in chloroform. The operating condition were as follows: name of column- RTS-5MS, diameter 30 cm, length 0.25mm,
temperature of the column- initial temperature 40˚C ( was held 2 min) , injector temperature- 220 ˚C, holding time 5 min, column packing-
column packing was done with 10% diethylene glycol succinate on 100-120 mesh diatomic CAW, splitting- samples were injected by splitting
Shamsun Naher et al / Int. J. Pharm. Phytopharmacol. Res. 2015; 5 (1): 35-40
37
with the spilt ratio 10, carrier gas- helium gas at constant pressure 90 kPa, sample dissolved- in chloroform, range of linear temperature
increase- 10˚C per min.
2.3 Preparation of fatty oil samples for GC-MS analysis
Fatty oil was diluted to 7% by chloroform. An inert gas (i.e. nitrogen) was introduced, from a large gas cylinder through the injection part, the
column and the detector. The flow rate of the carrier gas was adjusted to ensure reproducible retention time and to minimize detector dirt. The
sample was then injected by a micro syringe through a heated injection part when it was vaporized and carried into the column. The long tube
of the column was tightly packed with solid particles. The solid support was uniformly covered with a thin film of a high boiling liquid (the
stationary phase). The mobile and stationary phases were then partitioned by the samples and it was separated into the individual components.
The carrier gas and sample component was then emerging from the column and passed through a detector. The amount of each component as
concentration by the device and generates a signal which was registered electrically. The signal passed to a detector.
2.4 Identification of the components
The physicochemical properties of the fatty oil of Capsicum annuum Linn. (Dry chili) of two varieties from two different places of Bangladesh are
presented in Table-1. Interpretation of mass spectroscopy (GC-MS) was conducted using data base of National Institute Standard and
Technology (NIST) having more than 62000 patterns. The spectrum of the unknown component was compared with the spectrum of the known
component stored in the NIST library. The retention time, molecular weight, molecular formula and composition percentage of the sample
material was recorded and presented in Table -2 and Table-3 (the fatty oil of Capsicum annuum Linn. (Dry chili) of two varieties from two
different places of Bangladesh.)
Elemental analysis:
The mineral content was determined by EDXRF ANALYZER UNIQUANT (TM) STANDARDLESS METHOD in g/l00g dry weight basis of the
sample. The element analysis of Capsicum Annuum linn (dry chili) can be compared in different origins of Bangladesh.
Capsicum Annuum linn (dry chili) Mineral contents:
Mineral content such as K, Ca, Cl, S, Cu, Mn, Fe, Zn, Pb etc. were determined in its fresh Capsicum Annuum linn (dry chilli). Two varities of
Capsicum Annuum linn (dry chili) of different origin of Bangladesh like Dinajpur & chittagong were collected from local area. The result of
element analysis of its seed was determined by EDXRF ANALYZER UNIQUANT (TM) STANDARDLESS METHOD in g/l00g dry weight basis
of the sample. The result of minerals content of Capsicum Annuum linn (dry chili) from different origin of Bangladesh (Dinajpur and Chittagong)
appeared as element basis in Table-4.
3. RESULTS AND DISCUSSION
The result of the Physico-chemical properties of Capsicum annuum Linn. (Dry chili) fatty oil of Dinajpur and Chittagong appeared in Table -1.
The slight variation of this oil content and the composition of the fatty oil depend on several factors such genotype, stage of maturity, cultivation
peculiarities, soil composition and climate differences in various geographical locations. Fluctuation of the oil composition can import change in
the organoleptic properties of the plant belonging to the botanical spices and variety. So far we aware till now no systemic investigation on the
Capsicum annuum Linn. (Dry chili) have not been investigated in Bangladesh by using modern analytical techniques.
GC analyzed results which include the active principles with their retention time; molecular formula; molecular weight and composition of the
fatty oil of Capsicum annuum Linn. (Dry chili) of two varieties were presented in Table-2 and Table-3.
Total nine fatty acids were identified from the fatty oil of Dinajpur dry chili. The major constituents were 9, 12-Octadecadienoic acid (Z, Z)-
methyl ester, (44.64%), 10-Octadecanoic acid ,methyl ester (24.33%), Hexadecanoic acid ,methyl ester (17.40%).
On the other hand total ten fatty acids were identified from the fatty oil of Chittagong dry chili. The major constituents were Hexadecanoic acid
,methyl ester (30.15%), 9,12-Octadecadienoic acid(Z,Z)- methyl ester, (29.30%), 9-octadecenoic acid(z)-methyl ester(17.82%).
Results show that fatty oil from both of two areas oils are a complex mixture of numerous, many of which are found in trace amounts. It is worth
monitoring that there is a great variation in the chemical composition of these two region oil of Capsicum annuum Linn. (Dry chili). This confirms
that the reported variation in oil is due to geographic divergence and ecological conditions.
Shamsun Naher et al / Int. J. Pharm. Phytopharmacol. Res. 2015; 5 (1): 35-40
38
Table 1: Comparative studies on Physico-chemical properties of fatty oil of dry chili of Bangladesh (Dinajpur and Chittagong).
Physical properties
Dinajpur dry chili Chittagong dry chili
Oil yield (%) 9.36% 7.06%
Organoleptic
properties
Taste Spicy pungent taste Spicy pungent taste
Odor Spicy Spicy
Color Deep red Deep red
Appearance at room
temperature (30
0
c)
Homogeneous,
opaque liquid,
lighter than water
Homogeneous,
opaque liquid,
lighter than water
Ash 9.74% 6.27%
Moisture 16.33% 16.67%
Crude fiber 25.44% 24.99%
Protein
Carbohydrate
Food energy
Specific gravity at 30
0
c
Refractive index [ή
t c
]
3.37%
8.35%
377.85cal/g
0.92
1.47
3.11%
7.75%
419.40cal/g
0.91
1.49
Solubility in
Alcohol Soluble Soluble
Distilled water Insoluble Insoluble
Chloroform Soluble Soluble
CCl
4
Soluble Soluble
Pet-ether Soluble Soluble
Diethyle ether Soluble Soluble
n-Hexane Insoluble Insoluble
Chemical properties
Acid value 10.17 8.19
Saponification value 188.46 197.09
Iodine value 133.83 139.59
Table-2: Chemical constituents of the fatty oil of Dry chilly (from Dinajpur)
Peak No:
R
etention
Time
Area (
%)
Name of the Compound
Molecular
weight
Molecular
formula
1 17.634 2.72 Tetradecanoic acid,methyl ester 420.62 C
26
H
44
O
4
2 20.943 1.55 9-Hexadecanoic acid ,methyl ester 268.4348 C
17
H
32
O
2
3 21.440 17.40 Hexadecanoic acid ,methyl ester 270.4507 C
17
H
34
O
2
4 26.739 44.64 9,12-Octadecadienoic acid(Z,Z)- methyl ester, 294.47206 C
19
H
34
O
2
5 26.953 24.33 10-Octadecanoic acid ,methyl ester 296.4879 C
19
H
36
O
2
6 27.084 2.39 9-octadecenoic acid(z)-methyl ester 296.4879 C
19
H
36
O
2
7 27.874 3.76 Octadecanoic acid ,methyl ester 294.4721 C
19
H
34
O
2
8 33.346 1.43 Octadecanoic acid,9,10,12-trimethoxy-,methyl ester C
19
H
32
O
5
9 34.125 1.80 4-tert-butyl-2-methoxyquinoline 3-phenyl-2-piperidino-
2-propen-1-a 4-butyl-2,2-dimethyl-2H-1-benzopyran
Shamsun Naher et al / Int. J. Pharm. Phytopharmacol. Res. 2015; 5 (1): 35-40
39
Table-3: Chemical constituents of the fatty oil of Dry chilly (from Chittagong)
Peak No
.
Retention
Time
Area (
%)
Name of the
Compound
Molecular
weight
Molecular
formula
1 13.960 1.99 Dodecanoic
acid,methyl ester
C
12
H
24
O
2
2 17.638 5.69 Tetradecanoic
acid,methyl ester
420.62 C
26
H
44
O
4
3 20.943 1.14 9-Hexadecanoic acid
,methyl ester
268.4348 C
17
H
32
O2
4 21.464 30.15 Hexadecanoic acid
,methyl ester
270.4507 C
17
H
34
O
2
5 26.673 29.30 9,12-Octadecadienoic
acid(Z,Z)- methyl
ester,
294.47206 C
19
H
34
O
2
6 26.898 17.82 9-octadecenoic
acid(z)-methyl ester
296.4879 C
19
H
36
O
2
7 27.056 3.08 8-octadecenoic
acid,methyl ester
8 27.867 5.48 Octadecanoic acid
,methyl ester
294.4721 C
19
H
34
O
2
9 33.335 2.02 Octadecanoic
acid,9,10,12-
trimethoxy-,methyl
ester
10 34.139 3.34 Methoxylate ester of
methyl oleate
Table-4: Elemental analysis in percent comparatively (on dry matter basis) of dry chili from two different parts of Bangladesh (Dinajpur and
Chittagong).
S.
No
.
Component as
Element
Dry chili of Dinajpur
Dry chili of
Chittagong
1 K 85.16 82.54
2 Ca 6.44 5.26
3 Cl 5.31 8.89
4 S 1.62 1.79
5 Fe 0.713 0.500
6 Pb 0.322 0.646
7 Zn 0.258 0.137
8 Ti 0.0261 0.0609
9 Cu 0.0547 0.0476
10 Mn 0.0368 0.0709
11 Sr 0.0191 0.0257
12 Br 0.0081 0.0569
13 Ni - 0.0099
14 Se - 0.0040
Results of elemental analysis showed that percentage of minerals contained in fatty oils was varied in two different areas (Dinajpur and
Chittagong). K 85.16%, Ca 6.44%, S 1.62%, Fe 0.71% in Dinajpur fatty oil. Whereas, K 82.54%, Ca 5.26%, S 1.79%, Fe 0.500% in Chittagong
fatty oil.
Shamsun Naher et al / Int. J. Pharm. Phytopharmacol. Res. 2015; 5 (1): 35-40
40
4. ACKNOWLEDGEMENT
Authors are thankful to Abu Anis Jahangir, Director, Dhaka laboratories, Bangladesh Council of Scientific and Industrial Research (BCSIR).
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