International Food Research Journal 20(2): 601-605 (2013)

Journal homepage: http://www.ifrj.upm.edu.my

Consequence of fatty acids profile including trans fat in chocolate and pastry
samples

*1
Aftab, A. K., 2Sherazi, S.T.H, 1Rubina, S., 1Razia, S., 1Ambrat and 1Arfa, Y.
1
Pakistan Council of Scientific and Industrial Research Laboratories Complex (Applied
Chemistry Research Center) Karachi-75280, Pakistan
2
National Center of Excellence in Analytical Chemistry, University of Sindh, Jamshoro-
76080, Pakistan

Article history Abstract

Received: 5 June 2012 Due to reported detrimental health effects of diets which are high in trans fatty acids (TFA)
Received in revised form: particularly on blood lipids, two main convenience products (chocolate and pastry) were
6 November 2012
Accepted: 8 November 2012
investigated. In this regard, to determine fatty acids composition, with special emphasis on
trans fat content from extracted neat fat of the products, gas chromatography coupled with mass
Keywords spectroscopic technique was used. Total saturated and unsaturated fatty acid found were in the
range of 63.89- 84.35 and 15.24--36.11 for chocolate samples, and similarly 40.22-53.46% and
Chocolate
46.56-59.77% for pastry samples. The high amount of TFA was observed in all chocolate and
pastry
fatty acid profile pastry samples varied from 4.56-8.49 and 3.92-10.17% respectively.
trans fat
GC-MS © All Rights Reserved

Introduction to have a hypercholesterolemic effect. Pastry is the

name given to various kinds of baked goods e.g. small
Fats and oils of animal origin such as butter and cakes, tarts and other sweet baked goods, made from
lard are composed of saturated fatty acids (SFAs) is ingredients such as flour, milk, butter, shortening,
responsible for a number of cardiovascular diseases baking powder or eggs.
(Neaton and Wentworth, 1992). In order to reduce Usually vegetable oils that contain naturally
the saturated fat content of processed foods, the food occurring double bond are in cis configuration, prior
industry in developed countries moves progressively to hydrogenation; however, some double bonds are
from animal fat to vegetable fat sources (Tavella et in the trans configuration, so-called trans fatty acids
al., 2000). (TFAs) (Fritsche and Steinhart, 1998). Little amount
Unsaturated fats are heart-healthy, but they have of TFAs are also formed during deodorization process
some undesirable properties, specifically in air, they under high temperature (Sherazi et al., 2009). Elaidic
can gradually become rancid. Fat oxidation is the acid (C18:1t9) is the major trans fatty acid found in
main reason of deterioration of foods and can directly the processed fats. The amount of TFA in the diet is
affect many quality characteristics (Maache-Rezzoug of interest due to its adverse effects on human health,
et al., 1998). The shelf life of many type of food which include cardiovascular disease, abnormalities
depends on several parameters including: storage in infant development, coronary heart disease,
temperature and humidity, availability of oxygen in diabetes and inflammation (Ascherio et al., 1999).
the immediate environment (Nattress et al., 2004). These concerns pushed the Food and Agriculture
Main parameter’s to control and monitor the quality Organization (FAO) and the World Health
of chocolate and pastry include the amount of fat Organization to recommend that fats for human
and type of fatty acids especially trans fat contents consumption should contain less than 4% of the total
because the quality of fats plays a very important role fat as trans.
in food processing technology. In previous studies which are conducted for
Chocolate contains a variety of different the determination of trans fat in various types of
compounds such as saturated fat, polyphenols, foods, produced/sold in Pakistan, the trans fat
sterols, diterpenes, triterpenes, aliphatic alcohols and ranged in some foods were as follows: cooking and
methylxanthines (Knight, 2000). Because of its high partially hydrogenated oils 0.5–1.6 and 9.1–26.5%,
saturated fat content, chocolate is often postulated respectively (Sherazi et al., 2009), margarines (soft

*Corresponding author.
Email: chemist_afi@yahoo.com
602 Aftab et al./IFRJ 20(2): 601-605

and hard) and shortening 1.5–23.0 and 7.3–31.7%, was 1:50; helium was used as a carrier gas with a
respectively (Bhanger and Anwar, 2004), biscuits flow rate of 0.8 ml/min. The injector and detector
9.4–34.9%, (Kandhro et al., 2008) potato chips 0.4– temperatures were 240 and 260°C, respectively.
22.2% , doughnut cakes 5.3–17.1%, meat-containing The mass spectrometer was operated in the electron
foods (chicken thighs) 1.9–21.3%, and fried fish impact (EI) mode at 70 eV in the scan range of 50-
2.7–6.6% (Anwar et al., 2006). In Potato chips TFA 550 m/z.
ranged 4.91-14.13% (Kandhro et al., 2010). Cereal
based food 2.5% to 16.3% (Mahesar et al., 2010). No Calculations and statistical analyses
data has been found so far to know about fatty acid The identification of methyl esters carried out by
composition specially trans fat in Pakistani products NIST and Willy libraries. Two samples of each brand
such as chocolate and pastry. Having known about were collected and each sample was analyzed thrice
wide consumption of chocolates and pastry all over the and reported as mean ± Standard Deviation.
country as well as fats and oil are their raw material,
it was felt necessary to determine the saturated and Results and Discussion
trans fat in these products. It may lead to know about
serving of fats and trans fat by consumers and a data Applied chromatographic conditions allowed
base of fat and oil composition of products. analyzing fatty acid compositions, especially trans
fat composition of frequently consumed chocolate
Materials and Methods and pastry. The results of fatty acid composition of
analyzed chocolate and pastry samples are shown in
Samples Tables 1 and 2 respectively.
Six different chocolate and five puff pastry All analyzed chocolate and pastry samples were
samples were purchased from local supermarket found to contain the significant amount of saturated
in Jamshoro, Pakistan, and samples were coded as fatty acids (SFAs) such as, C8:0, C10:0, C12:0,
Chocolate (CHOC-1 to CHOC-6), and Pastry (PAS-1 C14:0, C15:0, C16:0, C17:0, C18:0 and C20:0.
to PAS-5). The selection of the brands was based Among SFAs C14:0, C16:0 and C18:0 were found
on highest vending amongst those available in the in large quantities in all the analyzed chocolate and
market. pastry samples. SFA contents ranged from 63.89
to 84.35% and from 40.22 to 53.46% in chocolate
Sample preparation and pastry samples, respectively. Figure 1 show
Lipid extraction from the samples was carried the representative chromatogram of different fatty
out with hexane under the operating conditions acids of analysed oil extracted from pastry sample,
specified in ICC Standard No. 136, and expressed chromatogram was in a good peak shape under the
as a percentage by mass of the product (ICC, 1982). optimized chromatographic conditions.
Fat obtained from samples was transferred into 5 ml Aro et al. (1997) has been reported the effect
glass vials. The decanted samples were all frozen at saturated fatty acids (SFAs) on human health,
-18°C until analysis. according the him SFAs with the chain length of
(C12:0–C16:0) carbon atoms cause atherogenic
Determination of fatty acid composition effects, while stearic acid (C18:0) has no effect
FAMEs were prepared using IUPAC standard and suppose as neutral, whereas oleic (C18:1) and
method 2.301 for the determination of FAC of the polyunsaturated fatty acids (PUFAs) produce a blood
chocolate and pastry samples (IUPAC, 1979). lipid lowering effect.
Table 1 and 2 represent the dominant fatty acid
GC-MS conditions among the saturated group is palmitic acid (C16:0)
The GC-MS analysis for FAMEs was performed and its range varied from 34.51 to 47.70 and 26.72 to
on Agilent 6890 N gas chromatography instrument 44.95% in chocolate and pastry samples respectively.
coupled with an Agilent MS-5975 inert XL mass The highest amount of palmitic acid e.g. 47.70% in
selective detector and an Agilent autosampler 7683-B CHOC-3 and 44.95% in PAS-5 was found. The amount
injector (Agilent Technologies, Little Fall, NY, USA). of palmitic acid indicates the greater contribution of
Highly polar, Rt-2560 biscyanopropyl siloxane palm oil in the manufacturing of chocolate and pastry
capillary column (100m x 0.25mm i.d x 0.2µm film products, similarly stearic acid (C18:0) was present
thickness) was used for the separation of fatty acid ranged from 17.87 to 30.41% and 6.12 to 11.52%.
methyl esters. The initial temperature of 150°C was Whereas, caprylic acid (C8:0) was present ranging
maintained for 2 min, raised to 230°C at the rate of from 0.11 to 0.75% and 0.03 to 1.67%; capric acid
4°C/min, and kept at 230°C for 5 min. The split ratio (C10:0) was present at 0.08 to 1.89%, 0.02 to 1.14%;
 Aftab et al./IFRJ 20(2): 601-605 603

Table 1. Saturated and unsaturated fatty acids composition (mean

percentage FAMEs) of chocolate samples
Samples- (n=3)
Fatty Acids Mean ± STD
CHOC-1 CHOC-2 CHOC-3 CHOC-4 CHOC-5 CHOC-6
C8:0 0.19 ±0.01 0.33 ±0.01 0.21 ±0.01 0.75 ±0.01 0.41 ±0.01 0.11 ±0.005
C10:0 1.81 ±0.05 1.67 ±0.05 0.08 ±0.001 1.08 ±0.05 1.59 ±0.05 1.89 ±0.06
C12:0 0.77 ±0.03 0.00 ±0.0 0.64 ±0.02 0.34 ±0.01 0.27 ±0.01 0.00 ±0.00
C14:0 7.03 ±0.25 8.69 ±0.25 3.94 ±0.15 7.73 ±0.25 11.05 ±0.45 4.18 ±0.15
C15:0 0.29 ±0.01 0.00 ±0.00 0.16 ±0.01 0.00 ±0.00 0.34 ±0.01 0.00 ±0.00
C16:0 34.51 ±0.55 44.60 ±1.25 47.70 ±1.25 43.14 ±1.25 40.71 ±1.25 44.86 ±1.25
C16:1n9c 0.00 ±0.00 0.15 ±0.01 0.00 ±0.00 0.00 ±0.00 4.17 ±0.15 4.43 ±0.18
C17:0 0.62 ±0.02 0.31 ±0.01 0.30 ±0.01 0.18 ±0.01 1.60 ±0.06 0.33 ±0.01
C18:0 17.87 ±0.55 20.82 ±0.25 25.63 ±0.58 28.98 ±1.25 26.60 ±1.25 30.41 ±1.24
C18:1n9trans 5.07 ±0.25 4.24 ±0.18 8.14 ±0.25 3.98 ±0.15 4.36 ±0.15 4.72 ±0.12
C18:1n9c 19.17 ±0.36 10.69 ±0.15 6.86 ±0.25 5.93 ±0.25 5.89 ±0.12 2.26 ±0.05
C18:2 n9,12t-t 0.33 ±0.01 0.78 ±0.03 0.35 ±0.01 2.62 ±0.08 0.20 ±0.01 0.68 ±0.02
C18:2 n9,12c-c 1.72 ±0.04 3.11 ±0.15 1.91 ±0.08 3.41 ±0.07 0.92 ±0.02 0.00 ±0.00
C18:2 n8,11c-c 6.62 ±0.15 3.33 ±0.15 4.01 ±0.15 0.00 ±0.00 0.00 ±0.00 0.64 ±0.03
C18:3 n9,12,15c-c-c 2.90 ±0.08 0.00 ±0.00 0.00 ±0.00 0.72 ±0.02 0.48 ±0.02 0.23 ±0.01 Figure 1. Representative chromatograph of fatty acid of pastry sample
C20:0 0.80 ±0.02 0.39 ±0.01 0.00 ±0.00 0.38 ±0.01 0.00 ±0.00 2.57 ±0.12
C20:1 n11c 0.30 ±0.01 0.20 ±0.01 0.00 ±0.00 0.17 ±0.01 0.00 ±0.00 2.28 ±0.11
analysed by GC-MS
Groups and ratio between the types of fatty acids from the composition of Chocolate samples
SFA 63.89 76.80 78.65 82.59 82.57 84.35
Total UFA 36.11 22.50 21.28 16.84 16.02 15.24
Total MUFA
ci MUFA
24.55
19.48
15.28
11.04
15.00
6.86
10.08
6.10
14.42
10.06
13.69
8.97
not present in the pastry samples.
Total PUFA 11.56 7.22 6.28 6.76 1.60 1.55 Oleic acid (C18:1 cis-9) was the major fatty acid
cisPUFA 11.23 6.44 5.93 4.14 1.40 0.87
Total TFA 5.40 5.02 8.49 6.60 4.56 5.40 present amongst the unsaturated fatty acids. Oleic
SFA/ UFA 1.77 3.41 3.70 4.90 5.15 5.54
cisMUFA+cisPUFA 30.71 17.48 12.79 10.24 11.46 9.84 acid is considered to be responsible for lowering the
cisPUFA/SFA
trans-FAs/ cis-FAs
0.18
0.18
0.08
0.29
0.08
0.66
0.05
0.64
0.02
0.40
0.01
0.55
LDL (bad) cholesterol levels (Kandhro et al., 2008).
SFA+TFA 69.29 81.83 87.14 89.19 87.14 89.75 Unsaturated fatty acids (UFA) ranged from 2.26
cisPUFA/(SFA+TFA) 0.16 0.08 0.07 0.05 0.02 0.01
cisMUFA+PUFA/SFA+TFA 0.44 0.21 0.15 0.11 0.13 0.11 to 19.17% and 31.72 to 43.90% in chocolate and
pastry samples respectively. The other members of
Table 2. Saturated and unsaturated fatty acids composition (mean monounsaturated fatty acids (MUFA), C16:1 cis-9
percentage FAMEs) of pastry samples and C20:1 cis-11 was also determined in chocolate
Fatty Acids
Samples- (n=3) Mean ± STD samples ranged from 0.15-4.43% and 0.17- 2.28%,
PAS-1 PAS-2 PAS-3 PAS-4 PAS-5
C8:0 1.67±0.05 0.13±0.01 0.06±0.003 0.03±0.001 0.00±0.00 respectively. While the pastry samples contained
C10:0
C12:0
1.14±0.05 0.36±0.01 0.09±0.003 0.14±0.01
1.21±0.04 0.25±0.01 0.30±0.01 0.00±0.00
0.02±0.001
0.70±0.02 only one member of MUFA, C20:1 n11 ranged from
C14:0
C16:0
2.63±0.12 1.37±0.05 3.38±0.12 3.56±0.15
26.72±0.74 42.43±1.27 31.55±1.25 31.41±1.25
0.37±0.01
44.95±1.24
0.08 to 1.57%.
C18:0 6.12±0.25 6.89±0.15 11.52±0.54 10.36±0.23 7.15±0.25 Polyunsaturated fatty acids (PUFAs) has major
C18:1n9trans 9.11±0.26 8.50±0.21 6.98±0.15 4.92±0.15 3.78±0.15
C18:1n9c 43.90±1.23 31.72±1.23 40.23±1.25 42.62±1.46 38.72±1.46 importance for biological and nutritional value;
C18:2 n9,12t-t 1.06±0.05 0.12±0.01 2.22±0.05 3.70±0.16 2.31±0.11
C18:2 n9,12c-c 0.00±0.00 5.08±0.18 0.35±0.01 0.16±0.01 0.14±0.005 C18:2 9,12 t–t; C18:2 cis-9,12; C18:2 cis-8,11 and
C18:2 n8,11c-c
C18:3 n9,12,15c-c-c
4.09±0.12 0.00±0.00 1.65±0.04 0.00±0.00
1.29±0.04 0.26±0.01 1.02±0.03 2.79±0.13
0.00±0.00
0.58±0.02 C18:3 cis-9,12,15, are the members of PUFA, ranged
C20:0
C20:1 n11c
0.73±0.02 1.32±0.03 0.16±0.01 0.39±0.01
0.33±0.01 1.57±0.05 0.64±0.02 0.08±0.004
0.26±0.01
1.03±0.05
from 0.20-2.62; 0.92-3.41; 0.64-6.62; 0.23-2.90%
Groups and ratio between the types of fatty acids from the composition of pastry samples determined in the chocolate samples respectively,
SFA 40.22 52.76 47.07 45.90 53.46
Total UFA 59.77 47.24 53.08 54.27 46.56 similarly, 0.12-1.06; 2.22-5.08; 1.65-4.09; 0.26-
Total MUFA 53.33 41.79 47.85 47.62 43.53
cisMUFA 44.22 33.29 40.87 42.70 39.75 2.79%, in pastry samples. PUFAs have beneficial
PUFA
cisPUFA
6.44
5.38
5.46
5.34
5.23
4.88
6.66
6.49
3.03
2.89 effects on both normal health and chronic diseases,
Total TFA
SFA/ UFA
10.17
3.96
8.61
6.12
7.33
6.42
5.08
9.03
3.92
13.62
such as regulation of lipid levels (Mori et al., 2000;
cisMUFA+cisPUFA
cisPUFA/SFA
49.61
0.13
38.63
0.10
45.75
0.10
49.19
0.14
42.64
0.05
Kris-Etherton et al., 2002).
Trans-FAs/ cis-FAs 0.20 0.22 0.16 0.10 0.09 The major trans fat observed in all analyzed
SFA+TFA 50.39 61.37 54.40 50.98 57.38
cis PUFA/(SFA+TFA) 0.11 0.09 0.09 0.13 0.05 samples was elaidic acid (C18:1 trans-9) ranged from
cisMUFA+PUFA/SFA+TFA 0.98 0.63 0.84 0.96 0.74
3.98 to 8.14% and 3.78 to 9.11% with mean values
5.08% and 6.66% in chocolate and pastry samples
lauric acid (C12:0) was present at 0.27 to 0.77% respectively. Sample (CHOC-3) contained highest
and 0.25 to 1.21%; myristic acid (C14:0) at 3.94 to amount (8.14%) of elaidic acid, while among the pastry
11.05% and 0.37 to 3.56%; arachidic acid (C20:0) samples PAS-1 contained highest amount (9.11%).
was present in range from 0.38 to 2.57% and 0.16 to In comparison with samples of Turkish chocolates
1.32% in chocolate and pastry samples respectively. ranged at 1.85-3.68% (Karabulut, 2007), and New
Some odd number fatty acids, expected from animal Zealand chocolate ranged at 0-3.1% (Saunders et al.,
fat source, like pentadecanoic acid (C15:0) (0.16- 2008). The analyzed chocolate samples contains high
0.34%) and margaric acid (C17:0) (0.18- 0.62%) amount of trans fat than those of reported values,
were also found in considerable amounts in chocolate however in comparison with Austrian pastry which
samples, however these odd number fatty acids were were ranged at flaky pastry 9.37–9.91%, Puff pastries
604 Aftab et al./IFRJ 20(2): 601-605

Table 3. Total fat content (approx. 5 g of sample), total gram/serving of used to express the nutritive value of edible oils and
fat and trans fat of chocolate and pastry samples
Total fat
fats (Alonso et al., 2000). The values of cis-PUFA/
Chocolate total fat Total trans
Samples
g/100g
Mean±STD1
g/serving 2 (%)
trans fat g/serving 3 total trans g/
100g 4
(SFA+TFA) were 0.01 and 0.16 in CHOC-6 and
CHOC-1 15.48±0.45 0.80 5.4 0.04 0.84 CHOC-1, while in pastry were at 0.05 and 0.13 in
CHOC-2 19.45±0.51 1.01 5.02 0.05
CHOC-3 25.14±1.25 1.31 8.49 0.11
0.98
2.13
PAS-5 and PAS-4, respectively.
CHOC-4 23.54±0.79 1.21 6.6 0.08 1.55 The level of total fat content, total fat g/serving
CHOC-5 29.52±1.25 1.51 4.56 0.07
CHOC-6 21.42±0.85 1.10 5.4 0.06
1.35
1.16
and trans fat g/serving present in oil samples obtained
Mean Values 5.91 0.07 5.16 1.16 1.33
from the chocolate and pastry samples were given in
Pastry Samples
PAS-1 36.52±1.25 1.88 10.17 0.19 3.714
Table 3. Total fat contents of the chocolate samples
PAS-2 30.15±1.24 1.54 8.61 0.13 2.596 ranged from 15.48-29.52% with a mean of 22.43%.
PAS-3 28.45±1.05 1.51 7.33 0.11 2.085
PAS-4 38.75±1.35 1.99 5.08 0.10 1.969 While in pastry samples ranged from 28.45-38.75%
PAS-5
Mean Values
29.48±0.95
7.02
1.54
0.12
3.92
5.19
0.06
1.69
1.156 with a mean of 32.67%. According to reported value
2.304
1
extracted fat ×100/weight of sample of fat contents (Wagner et al., 2008), of different
% of fat/100×weight of sample
types of pastries, like puff pastry or flaky pastry
2

3
% of trans/100× fat g/serving
% of trans/100× fat g/100g
contained fat from 26.8 to 38.87% (puff pastry) and
4

from 27.89 to 30.37% (flaky pastry). While total fat

0.10 to 6.57% (Wagner et al., 2008).
g/serving and total trans fats g/serving was ranged
Many countries have implemented rules to
between 0.80-1.51 and 0.04-0.11 with a mean value
keep controlled trans fat. Danish Government has
of 1.16 and 0.07g, in chocolate and pastry samples
prohibited the sale of food products containing >2%
respectively. Whereas among the pastry samples total
industrially produced TFA (Stender and Dyerberg,
fat g/serving and total trans fats g/serving ranged
2003). The American Governments have made rule
from 1.51 to 1.99 and 0.06 to 0.19, with mean value
that TFA levels should be indicated on food labels
of 1.69 and 0.12 g respectively. The results indicate
since 2003 (FDA, 2003). According to the nutritional
that 5.22 g of chocolate (CHOC-3) contain 1.31g of
recommendation the daily TFA intake should not
total fat and 0.11g/serving of trans fat (Table 3).
exceed 2.5–3 g/day (<1% of total energy) (Wagner
et al., 2008). Conclusion
Groups and ratio between the types of fatty acids
from the composition of chocolate and pastry samples In each of the observed categories, products with
were shown in Table 1 and 2, respectively. The SFA/ high levels of saturated and trans fat were found. The
UFA, show the relation between two major fatty acid higher amount of saturated and TFA demonstrated
groups of the extracted fat. Its value varies from 1.77 the quality of samples. Further, the amount of trans
to 5.54 (Chocolate) and 3.96 to 13.62 (Pastry). No fat was not mentioned on the label of any local
one sample had a ratio less than one, which clearly manufactured chocolate and pastry as there are no
indicates a high proportion of SFAs. The prevalence regulations of saturated and TFA in the Pakistan. In
of unsaturated over SFAs (smaller ratio) is considered order to reduce the intake of saturated and trans fat in
to be positive from the nutritional point of view. foods, the general healthy eating advice provided by
Mean value indices of cis-MUFA+cis-PUFA nutritionists should be followed.
ranged at (9.84 to 30.71 and 38.63 to 49.61) and cis-
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