Pemberian Ekastra Etanol Buah Buncis Untuk Menurunkan Kolesterol Total, LDL,dan HDL.
BEANS (PHASEOLUS VULGARIS L) EXTRACT AS ANTI DYSLIPIDEMIA:
Decrease of total cholesterol, malondialdehyde, low density lipoprotein, and increase of high
density lipoprotein on rat wistar
Sri Wahjuni
Chemistry Departement, Faculty of Math and Science Udayana University
ABSTRACT
Excessive fat consumption leads to increase of blood cholesterol level more than normal
condition which is called as Dyslipidemia. Dyslipidemia is an abnormal lipoprotein metabolism,
usually associated with lipoprotein over production or deficiency. Dyslipidemia is also often
described as an event of hyperlipidemia and also as a risk factor for cardiovascular disease.
This is an experimental study for accessing the ability of beans extract as an anti dislipidemia. The
anti dislipidemia were marked by decrease of total cholesterol, malondialdehid, Low Densyty Lipoprotein
levels, and increase HDL of Wistar rat induced dislipisemia with high cholesterol diet for 16 week. This
study applied randomized posttest only control group design. The samples were 24 Wistar Rat, randomized
into 6 treatment: negative control group diet standard (Treatment group 1), positive control group with diet
high cholesterol (Treatment cholesterol group 2), diet high cholesterol with beans ( Phaseolus vulgaris) 50
kg/bw (Treatment group 3),diet high cholesterol with beans (Phaseolus vulgaris L) Steenis) 100 kg/bw
(Tretment group 4), diet high cholesterol with beans (Phaseolus vulgaris L) 150 kg/bw (Treatment group 5).
diet drug simvastatin 0,18 mg/day /200 gram BB (Treatment group 6), After 16 weeks treatment, blood of
rats were driven for total cholesterol and MDA assays. LDL and HDL . All of data analyzed by Anova to
obtain the treatment different toward control by statistically with significance at α=0.05.
The result shows that extracts of beans (Phaseolus vulgaris L) in a dose of 150 mg/kg bw decrease
total cholesterol of 23.88 %, MDA of 70.60 % LDL 38.09 % also increase the HDL of 59,63%, Extract
beans (Phaseolus vulgaris L) has an ability to prevent cardiovascular disease. This is caused by the present
of phytosterol in the beans extract which has been phyto-chemistry tested and analyzed by GC-MS such as
stigmasterol substance.
Keywords: beans ( Phaseolus vulgaris L), dyslipidemia, Phytosterol, total cholesterol, LDL,HDL,MDA
INTRODUCTION
Nowdays many Indonesian people have the wrong diet, tend to like fast food that contains
nutrients that are not balanced. In general, this fast food containing saturated fat and high salt with
low fiber content. Excessive consumption of saturated fat, low carbohydrate, and less fiber from the
dailydietisafactorcausingincreasedbloodcholesterol.High blood cholesterol (dyslipidemia) is not
only experienced by obese people, but it can also occur in people who are thin and can afflict
people who are young. Various walks of life, have to try to live a healthy lifestyle in order to keep
cholesterol in the blood remained normal. In the body there is a normal cholesterol level is 160-200
mg (LIPI, 2009). Dilipidemia is a situation where an increase in blood cholesterol levels that exceed
normal circumstances, accompanied by increased levels of total cholesterol, triglycerides, and LDL
cholesterol. Dilipidemia occur when total cholesterol concentration reaches ≥ 240 mg / dl, LDL ≥
160 mg / dl, and triglycerides ≥ 150 mg / dl (Montgomery, 1993)
High blood cholesterol (dyslipidemia) is not only experienced by obese people, but it can also
occur in people who are thin and can afflict people who are young. Various walks of life, have to
try to live a healthy lifestyle in order to keep cholesterol in the blood remained normal. In the body
there is a normal cholesterol level is 160-200 mg (LIPI, 2009). Dilipidemia is a situation where an
increase in blood cholesterol levels that exceed normal circumstances, accompanied by increased
levels of total-cholesterol, triglycerides,and LDLcholesterol.Fat derived from food will undergo
pencernaaan processes in the intestine into free fatty acids, triglycerides, phospholipids and
cholesterol, then absorbed into the form of chylomicrons. Breakdown of chylomicrons circulate to
the rest of the liver and separated into cholesterol. Most cholesterol is discharged into the bile as
bile acids and in part again together with triglycerides to be allied with a particular protein
(apoprotein) and form very low density lipoprotein (VLDL). VLDL very low density lipoproteins
are further broken down by the enzyme lipoprotein LDL can not be the last 2-6 hours and
immediately be converted to LDL. This process is called mechanism of dyslipidemia (Soeharto, 2004).
.
Demage Lipids in LDL-cholesterol in the blood due to dyslipidemia produce various
products of decomposition are relatively stable, especially aldehyde reactive α, β-unsaturated, such
as malondialdehyde (MDA), 4-hydroxy-2-nonenal (HNE), heksanal, and 2-propenal (acrolein)
(Uchida, 2003 Carani.et.al, 2004). Malodialdehid a (marker) one of the products of decomposition
of per oksidadasi acids plural unsaturated fats (polyunsaturated fatty acids; PUFA).
Several studies medicinal plants tadisional that have been carried out include research conducted by
Ratni (2013), gedi (Abelmoschus manihot) shows the effect of hypolipidemic that can lower
cholesterol levels in male rats Wistar with the content of flavonoids and steroid in gedi
(Abelmoschus manihot) , Utariningsih (2007) determines the hair decoction of maize (Zea mays) is
effective in lowering cholesterol levels of mice, rats decreased levels of cholesterol using -sitosterol
compounds contained in the hair of corn (Zea mays).
According to experts the researchers herbal, black cumin (Nigella sativa) play a role in
reducing the excess cholesterol in the blood by active substance thymoquinone, omega 9, omega 3,
omega 6, saponins and phytosterols (Edi Junaedi, 2011).Subekti (2006) using leaf katuk (Sauropus
adrogynus L. Merr) to produce low cholesterol Japanese quail. Leaves katuk (Sauropus adrogynus
L. Merr) phytosterol-containing compounds that can lower cholesterol in Japanese quail products.
Wahyu (2013), using n-hexane extract of pomegranate fruit flesh is white (Punica granatum) which
can lower blood cholesterol levels in rats (Rattus norvegicus L.) with a content of phytosterols in
the flesh white pomegranate (Punica granatum). Based on research conducted by Jannah, et. al.
(2013), the fruit extract of beans (Phaseolus vulgaris L.) contain phytosterol compounds.
Phytosterols are natural plant sterols that contains a minimum of 70% -sitosterol. According
Bonsdoff-Nikander (2005), a compound phytosterols can lower cholesterol through competition
between cholesterol and phytosterols in a mixture of micelles, as well as by binding cholesterol in
the digestive tract. There has been no research on the effectiveness of the test pieces of bean
(Phaseolus vulgaris L.) in lowering cholesterol levels, so as to investigate the ability of ethanol
extract of the fruit of beans (Phaseolus vulgaris L.) as an anti-hypercholesterolemia through
reductions in total cholesterol, Low Density Lipoprotein (LDL) as well as an increase High Density
Lipoprotein (HDL) in Wistar rats dyslipidemia.
RESULT AND DISCUSSION
Results of the analysis by GC-MS techniques to extract ethanol kentak fruit beans (Phaseolus
vulgaris L.) obtained 19 peak compounds were detected , shown in Figure 1 with a retention time
(tR), peak area, and different molecular weights in Table 1.
Figure 1 Chromatograms of ethanol extractnof beans (Phaseolus vulgaris L.)
Table1. Substance detected in ethanol extract of beans
Peak
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
Conpound
Propane, 1-(1-ethoxydthoxy)-(CAS) 1-Ethoxy
Ethanol, 2,2-diethoxyEthane, 1,1-diethoxy-(CAS) 1,1-Diethoxyethane
Butane, 1,1-diethoxy-3-methyl1-ethoxy-1-pentoxy-ethane
1,1,3-Triethoxybutane
2-Pentanone, 5,5-diethoxyEthanol, 2,2-diethoxy-(CAS) Glycolaldehyde
1(2H)-Pyridinecarboxaldehyde, 3,4-dihydro-5-(2-piperidinyl)
Propanoic acid, 2-methyl-, 1-(1,1-dimethylethy)-2-methyl-1,3propanediyl ester (CAS)
1-(+)-Ascorbic acid 2,6-dihexadecanoate
Hexadecanoic acid, ethyl ester (CAS) Ethyl palmitate
9,12-Octadecadienoic acid (Z,Z)2H-Pyran, 2-(2-heptadecynyloxy)tetrahydro-(CAS)
Octadecanoic acid
9,12,15-Octadecatrienoic acid, ethyl ester, (Z,Z,Z)9-Octadecenamide, (Z)-(CAS) OLEOAMIDE
Hexadecanoic acid, 2-hydroxy-1-(hydrocymethyl)ethyl ester
(CAS)
Stigmasterol
tR
3,07
3,25
3,40
3,57
3,83
5,67
6,49
6,93
10,83
m/z
%area
45,00 6,63%
47,00 37,24%
45,00 22,92%
103,00 2,02%
73,00 2,68%
73,00 1,21%
103,00 1,43%
103,00 2,45%
110,00 0,78%
12,38
16,41
16,75
18,05
18,11
18,32
18,38
20,06
71,00
73,00
88,00
67,00
55,00
73,00
79,00
59,00
1,96%
5,16%
2,03%
4,88%
3,99%
1,78%
0,56%
0,61%
21,29
28,89
98,00
55,00
0,67%
0,96%
In the ethanol extract of the fruit of beans (Phaseolus vulgaris L.) contain phytosterol
compounds shown in the retention time of 28.89 minutes as stigmasterol. Results of mass spectra
of compounds stigmasterol the ethanol extract of the fruit of beans (Phaseolus vulgaris L.) is shown
in Fig.2, beheading pattern table shown in Table 2 and the results fragmentation stigmasterol
compounds shown in Figure 3.
Gambar 2. Spektra Massa Stigmasterol
Table 2 Pattern beheading
The possibility that the missing fragment
No m/z
1.
2.
3.
412
394
351
M+
M+-18
(M+-18)-43
-H2O
-C3H7
Figure 3. Fragmentation stigmasterol
Stigmasterol compound was detected at a retention time of 28.89 minutes and abundance
Of 0.96% showed the presence of molecular ions (M +) at m / z 412 were shot by high-energy
electrons (70ev) resulting fragment peaks at m / z 394 and releases H2O m / z 351 C3H7 release.
The molecular weight compounds l stigmasterol even showed that the compound does not contain
components of atoms N
DYSLIPIDEMIA
Decrease of total Cholesterol level
Measurement of total cholesterol was conducted to determine the amount of cholesterol in
the body. Results of the determination of total cholesterol shown in Table 3.
Table 3 Table average level of total cholesterol
Group
Average level of total-cholest (mg/dL)
K1
K2
P1
P2
P3
KS
62,50
88,75
69,55
68,50
67,55
69,85
Persentation
decrease
21,63%
22,81%
23,88%
21,29%
Average level of total-cholest
(mg/dL)
100
80
60
40
20
0
K1
K2
P1
P2
P3
KS
Figure 4. Average level of total cholesterol
Description:
K1 = negative control group; K2 = positive control group
P1 = group of ethanol extract of beans fruit 50 mg / kg bw
P2 = group of ethanol extract of fruit bean dose of 100 mg / kg bw
P3 = group of ethanol extract of fruit bean dose 150 mg / kg bw
KS = group of medication simvastatin dose of 0.18 mg / kg bw
Results of this study showed total cholesterol levels in the blood of Wistar rats
hypercholesterolemia decreased affected by the ethanol extract of the fruit of beans (Phaseolus
vulgaris L.) with dose variation is a dose of 50 mg / kg BW, 100 mg / kg bw and 150 mg / Kg BB.
Analysis of the results followed by one-way ANOVA test and Post Hoc Study by Tukey's
test / HSD to know the group that has the same effect or different from the others. ANOVA analysis
results which show the value of P
Decrease of total cholesterol, malondialdehyde, low density lipoprotein, and increase of high
density lipoprotein on rat wistar
Sri Wahjuni
Chemistry Departement, Faculty of Math and Science Udayana University
ABSTRACT
Excessive fat consumption leads to increase of blood cholesterol level more than normal
condition which is called as Dyslipidemia. Dyslipidemia is an abnormal lipoprotein metabolism,
usually associated with lipoprotein over production or deficiency. Dyslipidemia is also often
described as an event of hyperlipidemia and also as a risk factor for cardiovascular disease.
This is an experimental study for accessing the ability of beans extract as an anti dislipidemia. The
anti dislipidemia were marked by decrease of total cholesterol, malondialdehid, Low Densyty Lipoprotein
levels, and increase HDL of Wistar rat induced dislipisemia with high cholesterol diet for 16 week. This
study applied randomized posttest only control group design. The samples were 24 Wistar Rat, randomized
into 6 treatment: negative control group diet standard (Treatment group 1), positive control group with diet
high cholesterol (Treatment cholesterol group 2), diet high cholesterol with beans ( Phaseolus vulgaris) 50
kg/bw (Treatment group 3),diet high cholesterol with beans (Phaseolus vulgaris L) Steenis) 100 kg/bw
(Tretment group 4), diet high cholesterol with beans (Phaseolus vulgaris L) 150 kg/bw (Treatment group 5).
diet drug simvastatin 0,18 mg/day /200 gram BB (Treatment group 6), After 16 weeks treatment, blood of
rats were driven for total cholesterol and MDA assays. LDL and HDL . All of data analyzed by Anova to
obtain the treatment different toward control by statistically with significance at α=0.05.
The result shows that extracts of beans (Phaseolus vulgaris L) in a dose of 150 mg/kg bw decrease
total cholesterol of 23.88 %, MDA of 70.60 % LDL 38.09 % also increase the HDL of 59,63%, Extract
beans (Phaseolus vulgaris L) has an ability to prevent cardiovascular disease. This is caused by the present
of phytosterol in the beans extract which has been phyto-chemistry tested and analyzed by GC-MS such as
stigmasterol substance.
Keywords: beans ( Phaseolus vulgaris L), dyslipidemia, Phytosterol, total cholesterol, LDL,HDL,MDA
INTRODUCTION
Nowdays many Indonesian people have the wrong diet, tend to like fast food that contains
nutrients that are not balanced. In general, this fast food containing saturated fat and high salt with
low fiber content. Excessive consumption of saturated fat, low carbohydrate, and less fiber from the
dailydietisafactorcausingincreasedbloodcholesterol.High blood cholesterol (dyslipidemia) is not
only experienced by obese people, but it can also occur in people who are thin and can afflict
people who are young. Various walks of life, have to try to live a healthy lifestyle in order to keep
cholesterol in the blood remained normal. In the body there is a normal cholesterol level is 160-200
mg (LIPI, 2009). Dilipidemia is a situation where an increase in blood cholesterol levels that exceed
normal circumstances, accompanied by increased levels of total cholesterol, triglycerides, and LDL
cholesterol. Dilipidemia occur when total cholesterol concentration reaches ≥ 240 mg / dl, LDL ≥
160 mg / dl, and triglycerides ≥ 150 mg / dl (Montgomery, 1993)
High blood cholesterol (dyslipidemia) is not only experienced by obese people, but it can also
occur in people who are thin and can afflict people who are young. Various walks of life, have to
try to live a healthy lifestyle in order to keep cholesterol in the blood remained normal. In the body
there is a normal cholesterol level is 160-200 mg (LIPI, 2009). Dilipidemia is a situation where an
increase in blood cholesterol levels that exceed normal circumstances, accompanied by increased
levels of total-cholesterol, triglycerides,and LDLcholesterol.Fat derived from food will undergo
pencernaaan processes in the intestine into free fatty acids, triglycerides, phospholipids and
cholesterol, then absorbed into the form of chylomicrons. Breakdown of chylomicrons circulate to
the rest of the liver and separated into cholesterol. Most cholesterol is discharged into the bile as
bile acids and in part again together with triglycerides to be allied with a particular protein
(apoprotein) and form very low density lipoprotein (VLDL). VLDL very low density lipoproteins
are further broken down by the enzyme lipoprotein LDL can not be the last 2-6 hours and
immediately be converted to LDL. This process is called mechanism of dyslipidemia (Soeharto, 2004).
.
Demage Lipids in LDL-cholesterol in the blood due to dyslipidemia produce various
products of decomposition are relatively stable, especially aldehyde reactive α, β-unsaturated, such
as malondialdehyde (MDA), 4-hydroxy-2-nonenal (HNE), heksanal, and 2-propenal (acrolein)
(Uchida, 2003 Carani.et.al, 2004). Malodialdehid a (marker) one of the products of decomposition
of per oksidadasi acids plural unsaturated fats (polyunsaturated fatty acids; PUFA).
Several studies medicinal plants tadisional that have been carried out include research conducted by
Ratni (2013), gedi (Abelmoschus manihot) shows the effect of hypolipidemic that can lower
cholesterol levels in male rats Wistar with the content of flavonoids and steroid in gedi
(Abelmoschus manihot) , Utariningsih (2007) determines the hair decoction of maize (Zea mays) is
effective in lowering cholesterol levels of mice, rats decreased levels of cholesterol using -sitosterol
compounds contained in the hair of corn (Zea mays).
According to experts the researchers herbal, black cumin (Nigella sativa) play a role in
reducing the excess cholesterol in the blood by active substance thymoquinone, omega 9, omega 3,
omega 6, saponins and phytosterols (Edi Junaedi, 2011).Subekti (2006) using leaf katuk (Sauropus
adrogynus L. Merr) to produce low cholesterol Japanese quail. Leaves katuk (Sauropus adrogynus
L. Merr) phytosterol-containing compounds that can lower cholesterol in Japanese quail products.
Wahyu (2013), using n-hexane extract of pomegranate fruit flesh is white (Punica granatum) which
can lower blood cholesterol levels in rats (Rattus norvegicus L.) with a content of phytosterols in
the flesh white pomegranate (Punica granatum). Based on research conducted by Jannah, et. al.
(2013), the fruit extract of beans (Phaseolus vulgaris L.) contain phytosterol compounds.
Phytosterols are natural plant sterols that contains a minimum of 70% -sitosterol. According
Bonsdoff-Nikander (2005), a compound phytosterols can lower cholesterol through competition
between cholesterol and phytosterols in a mixture of micelles, as well as by binding cholesterol in
the digestive tract. There has been no research on the effectiveness of the test pieces of bean
(Phaseolus vulgaris L.) in lowering cholesterol levels, so as to investigate the ability of ethanol
extract of the fruit of beans (Phaseolus vulgaris L.) as an anti-hypercholesterolemia through
reductions in total cholesterol, Low Density Lipoprotein (LDL) as well as an increase High Density
Lipoprotein (HDL) in Wistar rats dyslipidemia.
RESULT AND DISCUSSION
Results of the analysis by GC-MS techniques to extract ethanol kentak fruit beans (Phaseolus
vulgaris L.) obtained 19 peak compounds were detected , shown in Figure 1 with a retention time
(tR), peak area, and different molecular weights in Table 1.
Figure 1 Chromatograms of ethanol extractnof beans (Phaseolus vulgaris L.)
Table1. Substance detected in ethanol extract of beans
Peak
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
Conpound
Propane, 1-(1-ethoxydthoxy)-(CAS) 1-Ethoxy
Ethanol, 2,2-diethoxyEthane, 1,1-diethoxy-(CAS) 1,1-Diethoxyethane
Butane, 1,1-diethoxy-3-methyl1-ethoxy-1-pentoxy-ethane
1,1,3-Triethoxybutane
2-Pentanone, 5,5-diethoxyEthanol, 2,2-diethoxy-(CAS) Glycolaldehyde
1(2H)-Pyridinecarboxaldehyde, 3,4-dihydro-5-(2-piperidinyl)
Propanoic acid, 2-methyl-, 1-(1,1-dimethylethy)-2-methyl-1,3propanediyl ester (CAS)
1-(+)-Ascorbic acid 2,6-dihexadecanoate
Hexadecanoic acid, ethyl ester (CAS) Ethyl palmitate
9,12-Octadecadienoic acid (Z,Z)2H-Pyran, 2-(2-heptadecynyloxy)tetrahydro-(CAS)
Octadecanoic acid
9,12,15-Octadecatrienoic acid, ethyl ester, (Z,Z,Z)9-Octadecenamide, (Z)-(CAS) OLEOAMIDE
Hexadecanoic acid, 2-hydroxy-1-(hydrocymethyl)ethyl ester
(CAS)
Stigmasterol
tR
3,07
3,25
3,40
3,57
3,83
5,67
6,49
6,93
10,83
m/z
%area
45,00 6,63%
47,00 37,24%
45,00 22,92%
103,00 2,02%
73,00 2,68%
73,00 1,21%
103,00 1,43%
103,00 2,45%
110,00 0,78%
12,38
16,41
16,75
18,05
18,11
18,32
18,38
20,06
71,00
73,00
88,00
67,00
55,00
73,00
79,00
59,00
1,96%
5,16%
2,03%
4,88%
3,99%
1,78%
0,56%
0,61%
21,29
28,89
98,00
55,00
0,67%
0,96%
In the ethanol extract of the fruit of beans (Phaseolus vulgaris L.) contain phytosterol
compounds shown in the retention time of 28.89 minutes as stigmasterol. Results of mass spectra
of compounds stigmasterol the ethanol extract of the fruit of beans (Phaseolus vulgaris L.) is shown
in Fig.2, beheading pattern table shown in Table 2 and the results fragmentation stigmasterol
compounds shown in Figure 3.
Gambar 2. Spektra Massa Stigmasterol
Table 2 Pattern beheading
The possibility that the missing fragment
No m/z
1.
2.
3.
412
394
351
M+
M+-18
(M+-18)-43
-H2O
-C3H7
Figure 3. Fragmentation stigmasterol
Stigmasterol compound was detected at a retention time of 28.89 minutes and abundance
Of 0.96% showed the presence of molecular ions (M +) at m / z 412 were shot by high-energy
electrons (70ev) resulting fragment peaks at m / z 394 and releases H2O m / z 351 C3H7 release.
The molecular weight compounds l stigmasterol even showed that the compound does not contain
components of atoms N
DYSLIPIDEMIA
Decrease of total Cholesterol level
Measurement of total cholesterol was conducted to determine the amount of cholesterol in
the body. Results of the determination of total cholesterol shown in Table 3.
Table 3 Table average level of total cholesterol
Group
Average level of total-cholest (mg/dL)
K1
K2
P1
P2
P3
KS
62,50
88,75
69,55
68,50
67,55
69,85
Persentation
decrease
21,63%
22,81%
23,88%
21,29%
Average level of total-cholest
(mg/dL)
100
80
60
40
20
0
K1
K2
P1
P2
P3
KS
Figure 4. Average level of total cholesterol
Description:
K1 = negative control group; K2 = positive control group
P1 = group of ethanol extract of beans fruit 50 mg / kg bw
P2 = group of ethanol extract of fruit bean dose of 100 mg / kg bw
P3 = group of ethanol extract of fruit bean dose 150 mg / kg bw
KS = group of medication simvastatin dose of 0.18 mg / kg bw
Results of this study showed total cholesterol levels in the blood of Wistar rats
hypercholesterolemia decreased affected by the ethanol extract of the fruit of beans (Phaseolus
vulgaris L.) with dose variation is a dose of 50 mg / kg BW, 100 mg / kg bw and 150 mg / Kg BB.
Analysis of the results followed by one-way ANOVA test and Post Hoc Study by Tukey's
test / HSD to know the group that has the same effect or different from the others. ANOVA analysis
results which show the value of P