Influence of β-Cyclodextrin on Reduction of Cholesterol Content in
Dairy, Egg and Meat Products
L.
1
Alonso
and J.
2
Fontecha
1Instituto
de Productos Lácteos de Asturias (CSIC)
2Instituto de Investigación en Ciencias de la Alimentación (CSIC-UAM)
INTRODUCTION
For the past 20 years, consumers have been reducing their fat consumption due to health concerns surrounding animal high fat diets. This trend has affected several
food products. Dairy, egg and meat products have been considered to increase the risk for cardiovascular diseases in humans because, in comparison to other lipid
sources, they contain a high proportion of cholesterol. Studies describing the association between dairy, egg and meat food consumption and the risk of
cardiovascular disease (CVD) have been inconsistent. In large epidemiological studies, consumption of high cholesterol fat products has been associated with an
increased CVD risk, while intake of low cholesterol fat foods has not. Methods for reducing cholesterol in foods have been developed, including between other,
adsorption with saponin and digitonin and removal by supercritical carbon dioxide extraction. Nowadays, a number of studies have indicated that cholesterol removal
from food products was most effectively achieved by beta-cyclodextrin (β-CD) (1).
OBJECTIVE
The aim of this work was the use of β-CD with pasteurized cow’s and ewe´s whole milk for making butter and cheese and its influence in cristal, natural eggs and
pates with the purpose of manufacturing low cholesterol food products.
MATERIAL AND METHODS
Analysis of cholesterol and fatty acids by gas chromatography
 Sampling
The technique chosen for cholesterol determination was as described by
Alonso et al. (2) using methylated fat by capillary gas chromatography (GC)
using a HP-5 fused silica capillary column 30 m x 0.32 mm i.d. x 0.25 m film
thickness. Approximately 30 mg anhydrous milk fat and 0.1 ml 5-α-cholestane
as internal standard (3.5 mg/mL in hexane) was dissolved in 1 mL of hexane;
0.5 μL of the resulting solution was injected for GC analysis. The GC analysis
was on an Agilent Technology 6890 chromatograph (Palo Alto, CA) equipped
with flame ionization detector (FID). Fatty acids were determined. as fatty
acid methyl esters on a Agilent Technology 6890 chromatograph (Palo Alto,
CA) with FID detector. Fatty acids were separated using CP-Sil 88 fused-silica
capillary column (3).
Raw cow´s and ewe´s pasteurized milk obtained from Central Lechera
Asturiana (Oviedo) and Montes de Toledo (Toledo) respectively, was treated
with β-CD (purity 99.5 %, Shandong Xinda Fine Chemical Co., Ltd. Qingdao,
China) as proposed by (1) for obtaining butter and cheese. Cream was churned
to butter using a continuous butter churn machine of 1500 Kg/h of butter
(Contimab, Westfalia, Denmark) and cheese was made with the protocol as
Manchego cheese. Crystal and natural egg were supplied by the company
Huevos Maryper (Murcia) and patés by Malvasia, S.A. (Soria).
 Lipid extraction
Statistical analysis
Lipids were extracted from samples following the International Standard
Method.
RESULTS AND DISCUSION
The cholesterol content (in %, g/100g fat) from control butter and β-CD
butter varied between 0.286 ± 0.02% and 0.015 ± 0.008% respectively with a
reduction of cholesterol after churning the cream for production of
commercial butter of 95.53 ± 2.8% (Figure 1). In cheese the cholesterol
reduction in three months Manchego cheese was 91.31 ± 2.4% (Figure 2)
comparing with the control cheese.
Figure 1
Figure 2
pA
pA
Cholesterol
35
35
5 a Cholestane
30
Cholesterol
5 a Cholestane
30
25
25
Control
With b -CD
20
Control
With b -CD
20
15
15
10
6
7
8
9
10
min
5.5
6
6.5
7
7.5
8
8.5
9
9.5
10
min
Cholesterol reduction in natural and crystal eggs ranged from 80 to 82 ± 2.6%
(Figure 3) and in paté (mouse) about 82.5 ± 2.3% (Figure 4) comparing with
control samples.
Figure 3
Figure 4
Experimental data were treated by analysis of variance (ANOVA) using the
statistical software SAS (version 8.02, SAS Institute Inc, Cary, NC, USA).
Differences among treatments were determined by statistical analysis using a
Student t-test where (P > 0.05) was considered statistically significant.
Table 1. Polyunsaturated fatty acids composition (g/100g fat) of
control milk and milk with β-cyclodextrin added.
Fatty acid
Control milk
ß-CD milk
Linoleic(n-6)
C18:2-cis-9, trans-13
C18:2-trans-8, cis-13
C18:2-cis-9, trans-12
C18:2-trans-11, cis-15
C18:2-cis-9,cis-15
C18:2-cis-9,cis-12
0.182 ± 0.05
0.053 ± 0.01
0.081 ± 0.01
0.433 ± 0.08
0.024 ± 0.01
1.836 ± 0.07
0.174 ±
0.050 ±
0.083 ±
0.412 ±
0.021 ±
1.819 ±
Linoleic conjugated (CLA)
C18:2 cis-9, trans-11
C18:2 cis-11, trans-13
C18:2 trans-10, cis-12
0.672 ± 0.08
0.009 ± 0.00
0.021 ± 0.01
0.663 ± 0.07
0.007 ± 0.01
0.019 ± 0.01
Linolenic
C18:3 (n-6)
C18:3 (n-3)
0.152 ± 0.01
0.315 ± 0.01
0.167 ± 0.01
0.332 ± 0.09
Total Conjugated linoleic
Total n-3 Polyunsaturated
Total n-6 Polyunsaturated
0.702 ± 0.08
0.441 ± 0.01
2.156 ± 0.10
0.689 ± 0.07
0.403 ± 0.02
2.142 ± 0.08
0.05
0.01
0.01
0.07
0.01
0.06
CONCLUSION
Control
With b-CD
Control
With b-CD
No difference (P < 0.05) for the linoleic acid C18:2 (n-6) for control milk
(1.836 ± 0.07) and treated β-CD milk (1.819 ± 0.06) was observed. In this
study, the three conjugated linoleic acid isomers analysed were C18:2-cis-9
trans-11, C18:2-cis-11 trans-13 and C18:2-trans 10 cis-12 (Table 1). The main
CLA isomer biologically active C18:2-cis-9 trans-11 (rumenic acid) did not show
differences (P < 0.05) between control milk (0.672 ± 0.08) and β-CD milk
(0.663 ± 0.07). The same patterns were observed for the total CLA in control
milk (0.702 ± 0.08) and treated milk (0.689 ± 0.07).
Results from the present study suggest that the treatment with β-cyclodextrin
can be applied to dairy, egg and meat products for making low cholesterol
food products without altering the nutritional fatty acid properties.
References
1. Alonso L, Cuesta P, Fontecha J, Juárez M and Gilliland SE. 2009. Use of β-cyclodextrin to
decrease the level of cholesterol in milk fat. J. Dairy Sci. 92: 863-860
2. Alonso, L., L. Lozada, J. Fontecha, and M. Juarez. 1995. Determination of cholesterol in
milk fat by gas chromatography with direct injection and sample saponification.
Chromatographia 41: 23-25
3. Alonso, L., J. Fontecha, L. Lozada, M.J.Fraga, and M. Juárez. 1999. Fatty acid
composition of caprine milk: major, branched-chain and trans fatty acids. J. Dairy Sci. 82:
878-884