Analisis Komponen Senyawa Kimia Minyak Atsiri Kulit Buah Asam Bunga (Citrus cf. nobilis Lour) Serta Uji Aktivitas Antibakteri Dan Antioksidan
71
LAMPIRAN
Universitas Sumatera Utara
72
Lampiran 1.Tanaman dan BuahAsam Bunga (Citrus cf. nobilis Lour)
1.1 Tanaman Asam Bunga (Citrus cf. nobilis Lour)
1.2 Buah Asam Bunga (Citrus cf. nobilis Lour)
Universitas Sumatera Utara
73
Lampiran 2. Hasil Uji Aktivitas Antioksidan Minyak Atsiri Kulit Buah Asam
Bunga
% Peredaman =
1.
1,094−1,002
1,094
x 100% = 8,40 %
1,094−0,935
1,094
x 100% = 14,53 %
Konsentrasi 30 ppm
% Peredaman =
4.
x 100%
Konsentrasi 20 ppm
% Peredaman =
3.
blanko
Konsentrasi 10 ppm
% Peredaman =
2.
blanko − sampel
1,094−0,883
1,094
x 100% = 19,28 %
Konsentrasi 40 ppm
% Peredaman =
1,094−0,832
1,094
x 100% = 23,94 %
Peredaman radikal bebas oleh minyak atsiri kulit buah asam bunga
Sampel
Absorbansi
% Peredaman
Blanko
1,094
-
10 ppm
1,002
8,40
20 ppm
0,935
14,53
30 ppm
0,883
19,28
40 ppm
0,832
23,94
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74
Lampiran 3. Perhitungan Nilai IC 50 Minyak Atsiri Kulit Buah Asam Bunga
X
0
10
20
30
40
Y
0,00
8,40
14,53
19,28
23,94
XY
0
84,0
290,6
578,4
957,6
X2
0
100
400
900
1600
∑X = 100
X = Konsentrasi
Y = % Perendaman
∑Y = 66,15
∑XY = 1910,6
∑X2 = 3000
a =
�(∑��)−(∑�)(∑�)
�(∑� 2 )–(∑�)2
=
=
=
5 (1910,6)− (100)(66,15)
5 (3000)−(100)2
9553−6615
15000 −10000
2938
5000
= 0,5876
b=
�∑� 2 �(∑�)−(∑�)(∑��)
�(∑� 2 )–(∑�)2
(3000)(66,15)−(100)(1910,6)
=
5 (3000)−(100)2
198450 −191060
=
=
15000 −10000
7390
5000
= 1,478
Jadi persamaan garis regresi Y = 0,5049X + 0,838
Nilai IC 50 :
50
= 0,5876 X + 1,478
0,5876 X
= 50 – 1,478
0,5876 X
= 48,522
X
IC 50
= 82,57
= 82,57=bp- ppm
Universitas Sumatera Utara
75
Lampiran 4. Hasil Data Kromatogram Hasil Analisa GC-MS Minyak Atsiri Kulit
Buah Asam Bunga
7
Chromatogram minyak as bunga 1 C:\GCMSsolution\Data\Project1\analisa as bunga\minyak as bunga1.qgd
R.Time
6.706
8.166
8.285
8.838
9.303
10.161
10.371
10.440
11.127
11.551
12.048
12.738
13.197
13.386
15.323
15.727
15.829
16.267
16.793
17.350
17.874
18.216
18.697
19.198
19.796
21.458
22.262
22.412
Area
1127495
1697541
14096001
1711258
2147807
6693231
79876032
6980228
267467
13454664
1104647
819417
2602799
551567
2654401
319763
427165
4379087
9822624
851112
252622
4016044
2551487
963006
3225504
305214
624796
3861812
Area%
0.64
0.96
7.95
0.97
1.21
3.77
45.05
3.94
0.15
7.59
0.62
0.46
1.47
0.31
1.50
0.18
0.24
2.47
5.54
0.48
0.14
2.26
1.44
0.54
1.82
0.17
0.35
2.18
30.0
TIC*1.00
36
35
32
34
29
30
31
33
28
26
20.0
27
23
24
20
21
10.0
Peak#
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
25
22
15
167
11
12
14 13
9
2
4
5
1
18
6 8
19
10
3
18,000,000
40.0
min
Peak Report TIC
Height Name
407639 .ALPHA.-PINENE, (-)- $$ Bicyclo[3.1.1]hept-2-ene, 2,6,6-trim
596839 Sabinene $$ Bicyclo[3.1.0]hexane, 4-methylene-1-(1-methylethy
4632104 l-.beta.-Pinene $$ Bicyclo[3.1.1]heptane, 6,6-dimethyl-2-methyl
578553 .beta.-Myrcene $$ 1,6-Octadiene, 7-methyl-3-methylene- (CAS)
684545 Octanal (CAS) n-Octanal $$ n-Octylal $$ Antifoam-LF $$ Octa
1953476 Benzene, methyl(1-methylethyl)- (CAS) Cymol $$ Cymene $$
17901825 l-Limonene $$ Cyclohexene, 1-methyl-4-(1-methylethenyl)-, (S)
1894115 1,8-Cineole $$ 2-Oxabicyclo[2.2.2]octane, 1,3,3-trimethyl- (CA
91637 Trans-Ocimene $$ 1,3,7-Octatriene, 3,7-dimethyl- (CAS) 2,6-Di
4315105 .gamma.-Terpinene $$ 1,4-Cyclohexadiene, 1-methyl-4-(1-meth
343388 1-Nonanol
262385 .ALPHA.-TERPINOLENE $$ Cyclohexene, 1-methyl-4-(1-met
829401 LINALOOL L $$
168566 Nonanal (CAS) n-Nonanal $$ n-Nonylaldehyde $$ Nonaldehyde
843111 CITRONELLA $$ 6-Octenal, 3,7-dimethyl- (CAS) Citronellal $
80673 MENTHOFURAN $$
93260 Isoborneol
1331261 l-4-Terpineol $$ 3-Cyclohexen-1-ol, 4-methyl-1-(1-methylethyl)
2978368 .ALPHA. TERPINEOL $$
241241 Decanal (CAS) n-Decanal $$ Decyl aldehyde $$ Decaldehyde $
74120 trans-Carveol $$ 2-Cyclohexen-1-ol, 2-methyl-5-(1-methylethen
1227934 .beta.-Citronellol $$ 6-Octen-1-ol, 3,7-dimethyl- (CAS) Citronel
801529 Z-Citral $$ 2,6-Octadienal, 3,7-dimethyl-, (Z)- (CAS) Neral $$ .
295810 GERANIOL $$
882547 E-Citral $$ 2,6-Octadienal, 3,7-dimethyl-, (E)- (CAS) Geranial $
102271 Farnesol $$ 2,6,10-Dodecatrien-1-ol, 3,7,11-trimethyl- (CAS) F
195853 .gamma.-Dodecalactone $$ 4-Hydroxydodecanoic acid lactone $
1005849 Evodone $$
Universitas Sumatera Utara
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Lampiran 4. Kondisi Alat GC-MS
Method [Comment]
===== Analytical Line 1 =====
[AOC-20i]
# of Rinses with Presolvent
# of Rinses with Solvent(post)
# of Rinses with Sample
Plunger Speed(Suction)
Viscosity Comp. Time
Plunger Speed(Injection)
Syringe Insertion Speed
Injection Mode
Pumping Times
Inj. Port Dwell Time
Terminal Air Gap
Plunger Washing Speed
Washing Volume
Syringe Suction Position
Syringe Injection Position
Use 3 Solvent Vial
[GC-2010]
Column Oven Temp.
Injection Temp.
Injection Mode
Flow Control Mode
Pressure
Total Flow
Column Flow
Linear Velocity
Purge Flow
Split Ratio
High Pressure Injection
Carrier Gas Saver
Splitter Hold
Oven Temp. Program
Rate
4.00
2.00
2
3
2
:High
:0.2 sec
:Middle
:High
:Normal
5
:0.3 sec
:No
:High
:8uL
:0.0 mm
:0.0 mm
:1 vial
:60.0 °C
:270.00 °C
:Split
:Pressure
:70.0 kPa
:62.4 mL/min
:1.16 mL/min
:39.4 cm/sec
:3.0 mL/min
:50.0
:OFF
:OFF
:OFF
Temperature(°C)
60.0
260.0
280.0
Hold Time(min)
5.00
10.00
5.00
< Ready Check Heat Unit >
Column Oven
: Yes
SPL1
: Yes
MS
: Yes
< Ready Check Detector(FTD) >
< Ready Check Baseline Drift >
< Ready Check Injection Flow >
SPL1 Carrier
: Yes
SPL1 Purge
: Yes
< Ready Check APC Flow >
< Ready Check Detector APC Flow >
External Wait
:No
Equilibrium Time
:3.0 min
[GC Program] [GCMSQP2010 Plus]
IonSourceTemp
:280.00 °C
Interface Temp.
:270.00 °C
Solvent Cut Time :0.00 min
Detector Gain Mode :Relative
Detector Gain
:0.00 kV
Threshold
1000
[MS Table]
--Group 1 - Event 1-Start Time
End Time
ACQ Mode
Event Time
:1.00min
:80.00min
:Scan
:0.50sec
Scan Speed
Start m/z
End m/z
769
:40.00
:400.00
Sample Inlet Unit
:GC
[MS Program]
Use MS Program
:OFF
Universitas Sumatera Utara
77
Lampiran 6 Determinasi Tumbuhan Asam Bunga
Universitas Sumatera Utara
78
Lampiran 7 Data Hasil Uji Antioksidan
Universitas Sumatera Utara
LAMPIRAN
Universitas Sumatera Utara
72
Lampiran 1.Tanaman dan BuahAsam Bunga (Citrus cf. nobilis Lour)
1.1 Tanaman Asam Bunga (Citrus cf. nobilis Lour)
1.2 Buah Asam Bunga (Citrus cf. nobilis Lour)
Universitas Sumatera Utara
73
Lampiran 2. Hasil Uji Aktivitas Antioksidan Minyak Atsiri Kulit Buah Asam
Bunga
% Peredaman =
1.
1,094−1,002
1,094
x 100% = 8,40 %
1,094−0,935
1,094
x 100% = 14,53 %
Konsentrasi 30 ppm
% Peredaman =
4.
x 100%
Konsentrasi 20 ppm
% Peredaman =
3.
blanko
Konsentrasi 10 ppm
% Peredaman =
2.
blanko − sampel
1,094−0,883
1,094
x 100% = 19,28 %
Konsentrasi 40 ppm
% Peredaman =
1,094−0,832
1,094
x 100% = 23,94 %
Peredaman radikal bebas oleh minyak atsiri kulit buah asam bunga
Sampel
Absorbansi
% Peredaman
Blanko
1,094
-
10 ppm
1,002
8,40
20 ppm
0,935
14,53
30 ppm
0,883
19,28
40 ppm
0,832
23,94
Universitas Sumatera Utara
74
Lampiran 3. Perhitungan Nilai IC 50 Minyak Atsiri Kulit Buah Asam Bunga
X
0
10
20
30
40
Y
0,00
8,40
14,53
19,28
23,94
XY
0
84,0
290,6
578,4
957,6
X2
0
100
400
900
1600
∑X = 100
X = Konsentrasi
Y = % Perendaman
∑Y = 66,15
∑XY = 1910,6
∑X2 = 3000
a =
�(∑��)−(∑�)(∑�)
�(∑� 2 )–(∑�)2
=
=
=
5 (1910,6)− (100)(66,15)
5 (3000)−(100)2
9553−6615
15000 −10000
2938
5000
= 0,5876
b=
�∑� 2 �(∑�)−(∑�)(∑��)
�(∑� 2 )–(∑�)2
(3000)(66,15)−(100)(1910,6)
=
5 (3000)−(100)2
198450 −191060
=
=
15000 −10000
7390
5000
= 1,478
Jadi persamaan garis regresi Y = 0,5049X + 0,838
Nilai IC 50 :
50
= 0,5876 X + 1,478
0,5876 X
= 50 – 1,478
0,5876 X
= 48,522
X
IC 50
= 82,57
= 82,57=bp- ppm
Universitas Sumatera Utara
75
Lampiran 4. Hasil Data Kromatogram Hasil Analisa GC-MS Minyak Atsiri Kulit
Buah Asam Bunga
7
Chromatogram minyak as bunga 1 C:\GCMSsolution\Data\Project1\analisa as bunga\minyak as bunga1.qgd
R.Time
6.706
8.166
8.285
8.838
9.303
10.161
10.371
10.440
11.127
11.551
12.048
12.738
13.197
13.386
15.323
15.727
15.829
16.267
16.793
17.350
17.874
18.216
18.697
19.198
19.796
21.458
22.262
22.412
Area
1127495
1697541
14096001
1711258
2147807
6693231
79876032
6980228
267467
13454664
1104647
819417
2602799
551567
2654401
319763
427165
4379087
9822624
851112
252622
4016044
2551487
963006
3225504
305214
624796
3861812
Area%
0.64
0.96
7.95
0.97
1.21
3.77
45.05
3.94
0.15
7.59
0.62
0.46
1.47
0.31
1.50
0.18
0.24
2.47
5.54
0.48
0.14
2.26
1.44
0.54
1.82
0.17
0.35
2.18
30.0
TIC*1.00
36
35
32
34
29
30
31
33
28
26
20.0
27
23
24
20
21
10.0
Peak#
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
25
22
15
167
11
12
14 13
9
2
4
5
1
18
6 8
19
10
3
18,000,000
40.0
min
Peak Report TIC
Height Name
407639 .ALPHA.-PINENE, (-)- $$ Bicyclo[3.1.1]hept-2-ene, 2,6,6-trim
596839 Sabinene $$ Bicyclo[3.1.0]hexane, 4-methylene-1-(1-methylethy
4632104 l-.beta.-Pinene $$ Bicyclo[3.1.1]heptane, 6,6-dimethyl-2-methyl
578553 .beta.-Myrcene $$ 1,6-Octadiene, 7-methyl-3-methylene- (CAS)
684545 Octanal (CAS) n-Octanal $$ n-Octylal $$ Antifoam-LF $$ Octa
1953476 Benzene, methyl(1-methylethyl)- (CAS) Cymol $$ Cymene $$
17901825 l-Limonene $$ Cyclohexene, 1-methyl-4-(1-methylethenyl)-, (S)
1894115 1,8-Cineole $$ 2-Oxabicyclo[2.2.2]octane, 1,3,3-trimethyl- (CA
91637 Trans-Ocimene $$ 1,3,7-Octatriene, 3,7-dimethyl- (CAS) 2,6-Di
4315105 .gamma.-Terpinene $$ 1,4-Cyclohexadiene, 1-methyl-4-(1-meth
343388 1-Nonanol
262385 .ALPHA.-TERPINOLENE $$ Cyclohexene, 1-methyl-4-(1-met
829401 LINALOOL L $$
168566 Nonanal (CAS) n-Nonanal $$ n-Nonylaldehyde $$ Nonaldehyde
843111 CITRONELLA $$ 6-Octenal, 3,7-dimethyl- (CAS) Citronellal $
80673 MENTHOFURAN $$
93260 Isoborneol
1331261 l-4-Terpineol $$ 3-Cyclohexen-1-ol, 4-methyl-1-(1-methylethyl)
2978368 .ALPHA. TERPINEOL $$
241241 Decanal (CAS) n-Decanal $$ Decyl aldehyde $$ Decaldehyde $
74120 trans-Carveol $$ 2-Cyclohexen-1-ol, 2-methyl-5-(1-methylethen
1227934 .beta.-Citronellol $$ 6-Octen-1-ol, 3,7-dimethyl- (CAS) Citronel
801529 Z-Citral $$ 2,6-Octadienal, 3,7-dimethyl-, (Z)- (CAS) Neral $$ .
295810 GERANIOL $$
882547 E-Citral $$ 2,6-Octadienal, 3,7-dimethyl-, (E)- (CAS) Geranial $
102271 Farnesol $$ 2,6,10-Dodecatrien-1-ol, 3,7,11-trimethyl- (CAS) F
195853 .gamma.-Dodecalactone $$ 4-Hydroxydodecanoic acid lactone $
1005849 Evodone $$
Universitas Sumatera Utara
76
Lampiran 4. Kondisi Alat GC-MS
Method [Comment]
===== Analytical Line 1 =====
[AOC-20i]
# of Rinses with Presolvent
# of Rinses with Solvent(post)
# of Rinses with Sample
Plunger Speed(Suction)
Viscosity Comp. Time
Plunger Speed(Injection)
Syringe Insertion Speed
Injection Mode
Pumping Times
Inj. Port Dwell Time
Terminal Air Gap
Plunger Washing Speed
Washing Volume
Syringe Suction Position
Syringe Injection Position
Use 3 Solvent Vial
[GC-2010]
Column Oven Temp.
Injection Temp.
Injection Mode
Flow Control Mode
Pressure
Total Flow
Column Flow
Linear Velocity
Purge Flow
Split Ratio
High Pressure Injection
Carrier Gas Saver
Splitter Hold
Oven Temp. Program
Rate
4.00
2.00
2
3
2
:High
:0.2 sec
:Middle
:High
:Normal
5
:0.3 sec
:No
:High
:8uL
:0.0 mm
:0.0 mm
:1 vial
:60.0 °C
:270.00 °C
:Split
:Pressure
:70.0 kPa
:62.4 mL/min
:1.16 mL/min
:39.4 cm/sec
:3.0 mL/min
:50.0
:OFF
:OFF
:OFF
Temperature(°C)
60.0
260.0
280.0
Hold Time(min)
5.00
10.00
5.00
< Ready Check Heat Unit >
Column Oven
: Yes
SPL1
: Yes
MS
: Yes
< Ready Check Detector(FTD) >
< Ready Check Baseline Drift >
< Ready Check Injection Flow >
SPL1 Carrier
: Yes
SPL1 Purge
: Yes
< Ready Check APC Flow >
< Ready Check Detector APC Flow >
External Wait
:No
Equilibrium Time
:3.0 min
[GC Program] [GCMSQP2010 Plus]
IonSourceTemp
:280.00 °C
Interface Temp.
:270.00 °C
Solvent Cut Time :0.00 min
Detector Gain Mode :Relative
Detector Gain
:0.00 kV
Threshold
1000
[MS Table]
--Group 1 - Event 1-Start Time
End Time
ACQ Mode
Event Time
:1.00min
:80.00min
:Scan
:0.50sec
Scan Speed
Start m/z
End m/z
769
:40.00
:400.00
Sample Inlet Unit
:GC
[MS Program]
Use MS Program
:OFF
Universitas Sumatera Utara
77
Lampiran 6 Determinasi Tumbuhan Asam Bunga
Universitas Sumatera Utara
78
Lampiran 7 Data Hasil Uji Antioksidan
Universitas Sumatera Utara