68

Lampiran A. Perhitungan Kandungan Protein Biji Melinjo

A.1 Standart BSA untuk Penentuan Total Protein Terlarut
Konsentrasi BSA
(g/mL)
0

Hasil Absorbansi

0.043

0.169

10

0.349

15

0.484

20

0.553

Kurva Standart BSA

25

konsentrasi (g/L)

20

y = 33.34x - 0.213
R = 0.977

15
10
5
0
-5 0

0.1

0.2
0.3
0.4
Hasil absorbansi

0.5

0.6

A.2 Hasil Pengukuran Konsentrasi Protein Terlarut

SAMPEL

ABSORBANSI

KONSENTRASI
RERATA
VOLUME
(g/L)
KONSENTRASI
(mL)
U1
U2
U3
4.63
4.71
4.63
4.66 0.05
42,5

JUMLAH
PROTEIN
(mg)

Gg-PK

U1
0.700

U2
0.713

U3
0.701

Gg-PI

0.852

0.882

0.914

5.64

5.84

6.05

5.84 0.21

12,5

73

Gg-PH

0.437

0.439

0.439

2.87

2.88

2.88

2.88 0,01

6,4

18,4

197,9

69

Lampiran B. Penghambatan Aktivitas Enzim Alfa Amilase dan Alfa Glukosidase

B.1 Perhitungan aktivitas penghambatan Alfa Amilase
SAMPEL

U1
Gg-PK 0.530
Gg-PI
0.595
Gg-PH
0.524
Akarbose 0.067
0.449
K+
0.277
K0.172

S+
U2
0.535
0.597
0.519
0.065

U3
0.538
0.598
0.526
0.062

U1
0.432
0.458
0.477
0.062

SU2
0.438
0.456
0.471
0.059

U3
0.446
0.455
0.477
0.059

S(+) - S (-)
U1
U2 U3
0.098 0.097 0.092
0.137 0.141 0.143
0.047 0.048 0.049
0.005 0.006 0.003

rerata
STDEV
(%)

Penghambatan (%)
43.023
20.349
72.674
97.093

43.605
18.023
72.093
96.512

46.512
16.860
71.512
98.256

44.38
18.41
72.09
97.29

1.87
1.78
0.58
0.89

B.2 Perhitungan aktivitas penghambatan Alfa Glukosidase

S+
SAMPEL

U1

U2

SU3

U1

U2

S(+) - S (-)
U3

U1

U2

U3

Penghambatan (%)

rerata
STDEV
(%)

Gg-PK

0.754

0.763

0.753

0.375

0.39

0.396

0.379

0.373

0.357

27.8

29.0

32.0

29.6

2.2

Gg-PI

0.779

0.797

0.78

0.33

0.335

0.314

0.449

0.46

0.466

14.5

12.0

11.2

12.6

1.7

Gg-PH

0.695

0.7

0.712

0.626

0.627

0.628

0.069

0.07

0.084

86.9

86.1

84.0

85.7

1.5

Akarbose

0.051

0.051

0.051

0.037

0.036

0.034

0.014

0.02

0.017

97.3

97.1

96.8

97.1

0.3

0.746

0.746

0.746

0.213

0.22

0.229

0.533

0.526

0.517

70

Lampiran C. Perhitungan Derajat Hidrolisis

C.1 Standart derajat hidrolisis dengan L-Leucin
konsentrasi LLeucin (L)
0

absorbansi

Kurva Standart Derajat Hidrolisis L-Leucin

0.45
0.4

0.117

0.35

0.156

0.205

0.267

0.331

0.401

Nilai Absorbansi

y = 0.057x + 0.045
R = 0.990

0.3
0.25
0.2

0.15
0.1
0.05
0
0

Konsentrasi L-Leucin

71

C.2 Hidrolisis asam Gg-PI

Tujuan : memotong seluruh ikatan peptida
Prinsip : sampel + HCl konsentrasinya 6 N HCl
Langkah : 12,08 N HCl diencerkan menjadi 6 N HCl
M1 x V1 = M2 x V2
12,08 N x V1 = 6 N x 500 L
V1 = 248,3 L
Jadi 12,08 N HCl diambil sebanyak 248,3 L dan ditambahkan sampel Gg-PI
sebanyak 251,7 L, kemudian diinkubasi 1100C selama 12 jam. Hasil hidrolisis GgPI secara asam disebut sebagai Gg-PI(asam).
D.3 Perhitungan derajat hidrolisis

Konsentrasi Gg-PI = 5,84 g/ L

Total asam -amino Gg-PI yang dihidrolisis secara asam (Gg-PI(asam))

248,3 L Gg-PI + 251,7 L 12,08 N HCl = 500 L Gg-PI(asam) dengan

jumlah protein yang dimasukkan 1,45 mg.

Gg-PI(asam)

yang

diencerkan

7x

untuk

pengukuran

-amino

menggunakan metode TNBS (10 L Gg-PI(asam) dalam 60 L

aquadest) = Gg-PIasam 7x

Absorbansi 25 L Gg-PIasam 7x pada 420 (TNBS) = 0.512 sehingga

jumlah asam -amino dalam 25 L Gg-PIasam 7x = 8,44 g

Total asam -amino terlarut sebesar 59.08 g/146 mg protein atau

404,6 mg/g protein.

Total asam -amino terlarut Gg-PH

Gg-PH diencerkan 2x untuk pengukuran -amino menggunakan

metode TNBS (10 L Gg-PH2x pada 420 (TNBS)= Gg-PH2x

72

Absorbansi 25 L Gg-PHasam2x pada 420 (TNBS) = 0.524 sehingga

jumlah asam -amino dalam 25 L Gg-PHasam2x = 8,65 g

Total asam -amino terlarut sebesar 17.3g/72 mg protein atau 240,2

mg/g protein.

Perhitungan derajat hidrolisis (DH)

DH =

404,6240,2
404,6

100% = 40,6 %

73

Lampiran D. Pengujian Statistik Data Penelitian

D.1 Uji Normalitas Data
One-Sample Kolmogorov-Smirnov Test

N
Normal Parameters a,b
Most Extreme
Dif f erences

Aktiv it as
enzim
12
58,0426
30,8960
,169
,146
-,169
,584
,885

Mean
St d. Dev iation
Absolute
Positiv e
Negativ e

Kolmogorov -Smirnov Z
Asy mp. Sig. (2-tailed)
a. Test distribution is Normal.
b. Calculated f rom data.

D.2 Uji Homogenitas Levene-Statistik

Enzim Alfa Amilase
Test of Homogeneity of Variances
Aktiv it as enzim
Lev ene
St at ist ic
2,150

df 1

df 2
3

Sig.
,172

Enzim Alfa Glukosidase

Test of Homogeneity of Variances
Alpha glukosidase
Lev ene
St at ist ic
2,585

df 1

df 2
3

Sig.
,126

Alpha
glukosidase
12
56,2222
37,5017
,271
,241
-,271
,937
,343

74

D.3 Uji One Way ANOVA

One Way Anova Parameter Enzim Alfa Amilase
Descriptives
Aktiv itas enzim

N
Gg-PK
Gg-PI
Gg-PH
Akarbose
Total

3
3
3
3
12

Mean
44,380
18,411
72,093
97,287
58,043

Std.
Deviation Std. Error
1,869
1,079
1,776
1,025
,581
,336
,888
,513
30,896
8,919

95% Confidence
Interv al for Mean
Lower
Upper
Bound
Bound
39,737
49,023
13,999
22,823
70,649
73,537
95,081
99,493
38,412
77,673

Minimum
43,02
16,86
71,51
96,51
16,86

Maximum
46,51
20,35
72,67
98,26
98,26

ANOVA
Aktiv it as enzim

Between Groups
Within Groups
Total

Sum of
Squares
10484,609
15,549
10500,158

df
3
8
11

Mean Square
3494,870
1,944

F
1798,126

Sig.
,000

75

Uji LSD Parameter Enzim Alfa Amilase

Multiple Comparisons
Dependent Variable: Aktiv itas enzim
LSD

(I) Perlakuan
Gg-PK

Gg-PI

Gg-PH

Akarbose

(J) Perlakuan
Gg-PI
Gg-PH
Akarbose
Gg-PK
Gg-PH
Akarbose
Gg-PK
Gg-PI
Akarbose
Gg-PK
Gg-PI
Gg-PH

Mean
Diff erence
(I-J)
Std. Error
25,9690*
1,1383
-27,7132*
1,1383
-52,9070*
1,1383
-25,9690*
1,1383
-53,6822*
1,1383
-78,8760*
1,1383
27,7132*
1,1383
53,6822*
1,1383
-25,1938*
1,1383
52,9070*
1,1383
78,8760*
1,1383
25,1938*
1,1383

Sig.
,000
,000
,000
,000
,000
,000
,000
,000
,000
,000
,000
,000

95% Confidence Interv al

Lower Bound Upper Bound
23,3440
28,5939
-30,3381
-25,0882
-55,5319
-50,2820
-28,5939
-23,3440
-56,3071
-51,0572
-81,5009
-76,2510
25,0882
30,3381
51,0572
56,3071
-27,8187
-22,5689
50,2820
55,5319
76,2510
81,5009
22,5689
27,8187

*. The mean diff erence is signif icant at the .05 lev el.

One Way Anova Parameter Alfa Glukosidase

Descriptives
Alpha glukosidase

N
Gg-PK
Gg-PI
Gg-PH
Akarbose
Total

3
3
3
3
12

Mean
29,587
12,571
85,651
97,079
56,222

Std.
Deviation Std. Error
2,166
1,251
1,693
,977
1,480
,854
,291
,168
37,502
10,826

95% Confidence
Interv al for Mean
Lower
Upper
Bound
Bound
24,206
34,968
8,366
16,777
81,975
89,326
96,357
97,802
32,395
80,050

Minimum
27,81
11,24
84,00
96,76
11,24

Maximum
32,00
14,48
86,86
97,33
97,33

76

ANOVA
Alpha glukosidase

Between Groups
Within Groups
Total

Sum of
Squares
15450,473
19,664
15470,138

df
3
8
11

Mean Square
5150,158
2,458

F
2095,221

Sig.
,000

Uji LSD Parameter Alfa Glukosidase

Multiple Comparisons
Dependent Variable: Alpha glukosidase
LSD

(I) Perlakuan
Gg-PK

Gg-PI

Gg-PH

Akarbose

(J) Perlakuan
Gg-PI
Gg-PH
Akarbose
Gg-PK
Gg-PH
Akarbose
Gg-PK
Gg-PI
Akarbose
Gg-PK
Gg-PI
Gg-PH

Mean
Diff erence
(I-J)
Std. Error
17,0159*
1,2801
-56,0635*
1,2801
-67,4921*
1,2801
-17,0159*
1,2801
-73,0794*
1,2801
-84,5079*
1,2801
56,0635*
1,2801
73,0794*
1,2801
-11,4286*
1,2801
67,4921*
1,2801
84,5079*
1,2801
11,4286*
1,2801

*. The mean diff erence is signif icant at the .05 lev el.

Sig.
,000
,000
,000
,000
,000
,000
,000
,000
,000
,000
,000
,000

95% Confidence Interv al

Lower Bound Upper Bound
14,0639
19,9678
-59,0154
-53,1115
-70,4440
-64,5401
-19,9678
-14,0639
-76,0313
-70,1274
-87,4599
-81,5560
53,1115
59,0154
70,1274
76,0313
-14,3805
-8,4766
64,5401
70,4440
81,5560
87,4599
8,4766
14,3805