Lampiran 1.a Data Kadar Air Kelopak Rosella Kadar air (%) = kehilangan berat (g) x 100 Sampel sebelum kering (g)
Kehilangan berat = berat sampel mula-mula – berat sampel setelah dikeringkan
Kadar air (%) = 2- 0,38 x 100 2
= 81%
Lampiran 1.b Data nilai pH ekstrak pH awal Kondisi Ekstraksi Ulangan pH
70OC 15 menit 1 2,85 2 2,85 70OC 30 menit 1 2,85 2 2,85 85OC 15 menit 1 2,87 2 2,94 85OC 30 menit 1 2,80 2 2,90 100OC 15 menit 1 2,79 2 2,77 100OC 30 menit 1 2,78 2 2,94
63
Lampiran 2. Uji Anova nilai pH
Univariate Analysis of Variance
Tests of Between-Subjects Effects Dependent Variable:pH
Source
Type III Sum of
Squares df Mean Square F Sig.
Corrected Model .016a 5 .003 .941 .516 Intercept 97.413 1 97.413 2.858E4 .000 Waktu .000 1 .000 .061 .813 Suhu .007 2 .003 .971 .431 waktu * suhu .009 2 .005 1.352 .328 Error .020 6 .003 Total 97.449 12 Corrected Total .036 11
Lampiran 3. Data total fenol Kurva standar
Kondisi
Ekstraksi Ulangan Absorbansi
Konsentrasi (ppm)
Konsentrasi ( mg GAE/g
rosella basah ) Rata-rata
70OC 15 menit 1 0,364 105,93 0,49 0,49 2 0,364 105,93 0,49 70OC 30 menit 1 0,404 115,02 0,53 0,53 2 0,405 115,25 0,53 85OC 15 menit 1 0,426 120,02 0,55 0,57 2 0,452 125,93 0,58 85OC 30 menit 1 0,404 112,75 0,52 0,55 2 0,459 127,52 0,59 100OC 15 menit 1 0,491 134,80 0,62 0,59 2 0,436 122,30 0,56 100OC 30 menit 1 0,544 146,84 0,68 0,67 2 0,543 146,61 0,67
Contoh perhitungan total fenol
X : konsentrasi asam galat; y : absorbansi Y = 0,0044x- 0,1021
0,364= 0,0044x – 0,1021 X= 105,93 ppm
Konsentrasi (mg GAE/g) = 105,93 mg asam galat x 230 ml 1000 ml ekstrak 50g = 0,49 mg GAE/g y = 0,0044x - 0,1021 R² = 0,9902 A b sor b ansi Konsentrasi (ppm)
Kurva Standar Larutan
Asam Galat
65
Lampiran 4. Uji Anova total fenol
Univariate Analysis of Variance
Tests of Between-Subjects Effects Dependent Variable:fenol
Source
Type III Sum of
Squares df Mean Square F Sig.
Corrected Model .039a 5 .008 9.922 .007 Intercept 3.865 1 3.865 4.882E3 .000 Waktu .004 1 .004 5.568 .056 Suhu .030 2 .015 19.168 .002 waktu * suhu .005 2 .002 2.853 .135 Error .005 6 .001 Total 3.909 12 Corrected Total .044 11
a. R Squared = ,892 (Adjusted R Squared = ,802)
Post Hoc Tests Suhu Homogeneous Subsets Fenol Duncan suhu N Subset 1 2 3 70 4 .5100 85 4 .5600 100 4 .6325 Sig. 1.000 1.000 1.000
Means for groups in homogeneous subsets are displayed. Based on observed means.
ANOVA Fenol
Sum of Squares df Mean Square F Sig.
Between Groups .039 5 .008 9.922 .007
Within Groups .005 6 .001
Total .044 11
Post Hoc Tests Interaksi Homogeneous Subsets Fenol Duncan interaks i N
Subset for alpha = 0.05
1 2 3 1 2 .4900 2 2 .5300 .5300 4 2 .5550 .5550 3 2 .5650 5 2 .5900 6 2 .6750 Sig. .067 .089 1.000
67
Lampiran 5. Data total asam tertitrasi Kondisi ekstraksi
sampel Ulangan Berat sampel (g) ml 0,1 NaOH TAT (%) Rata-rata pH
70OC 15 menit 1 10,078 0,8 6,569 6,63 2,85 2 10,107 0,8 6,699 2,85 70OC 30 menit 1 10,098 0,7 5,404 5,43 2,85 2 10,109 0,7 5,449 2,85 85OC 15 menit 1 10,1 0,7 5,642 5,74 2,84 2 10,008 0,7 5,839 2,87 85OC 30 menit 1 10,026 0,7 5,600 5,57 2,8 2 10,102 0,7 5,532 2,9 100OC 15 menit 1 10,009 0,8 6,458 6,23 2,77 2 10,099 0,8 6,007 2,79 100OC 30 menit 1 10,034 1,1 9,130 9,06 2,94 2 10,106 1,1 8,986 2,78
Contoh perhitungan total asam tertitrasi:
Total asam tertitrasi = ml 0,1 NaOH x N NaOH x Faktor Pengenceran x 100% gram ekstrak
6,569% = 0,8 x 0,08 x 10 x 100% 10,078
Lampiran 6. Uji Anova total asam tertitrasi
Univariate Analysis of Variance Tests of Between-Subjects Effects Dependent Variable:totalasam
Source
Type III Sum of
Squares Df Mean Square F Sig.
Corrected Model 18.460a 5 3.692 154.697 .000 Intercept 498.199 1 498.199 2.087E4 .000 Waktu .691 1 .691 28.961 .002 Suhu 8.985 2 4.493 188.236 .000 waktu * suhu 8.784 2 4.392 184.025 .000 Error .143 6 .024 Total 516.802 12 Corrected Total 18.604 11
a. R Squared = ,992 (Adjusted R Squared = ,986)
Post Hoc Tests suhu Homogeneous Subsets Duncan Suhu N Subset 1 2 3 85 4 5.6525 70 4 6.0300 100 4 7.6475 Sig. 1.000 1.000 1.000
Means for groups in homogeneous subsets are displayed. Based on observed means.
69
Waktu Dependent Variable:total asam tertitrasi
waktu Mean Std. Error
95% Confidence Interval Lower Bound Upper Bound
15 6.203 .063 6.049 6.358
30 6.683 .063 6.529 6.838
ANOVA Totalasam
Sum of Squares df Mean Square F Sig.
Between Groups 18.460 5 3.692 154.697 .000
Within Groups .143 6 .024
Total 18.604 11
Post Hoc Tests Interaksi Homogeneous Subsets Duncan kombinasi N Subset 1 2 3 4 70*30 2 5.4250 85*30 2 5.5650 85*15 2 5.7400 100*15 2 6.2350 70*15 2 6.6350 100*30 2 9.0600 Sig. .096 1.000 1.000 1.000
Means for groups in homogeneous subsets are displayed. Based on observed means.
Lampiran 7.a. Data inhibisi ekstrak rosella pH awal terhadap enzim α-amilase
Bacillus sp
Kondisi Ekstraksi Ulangan Inhibisi (%) Rata-rata (%)
700C, 15mnt 1 95,67 94,72 93,78 2 93,23 92,90 92,56 700C, 30mnt 1 95,13 94,32 93,51 2 93,91 91,54 89,18 850C, 15mnt 1 98,65 97,63 96,62 2 94,59 96,96 99,32 850C, 30mnt 1 93,23 88,50 83,76 2 99,32 99,66 100,00 1000C, 15mnt 1 92,56 95,60 98,65 2 93,23 91,20 89,18 1000C, 30mnt 1 99,32 99,66 100,00 2 95,26 97,63 100,00
71
Lampiran 7.b. Data inhibisi ekstrak rosella pH saluran pencernaan terhadap enzim α-amilase Bacillus sp
Kondisi ekstraksi Ulangan Inhibisi (%) Rata-rata (%)
700C, 15mnt 1 37,28 36,95 36,63 2 42,29 43,83 45,37 700C, 30mnt 1 8,74 8,29 7,84 2 8,74 8,68 8,61 850C, 15mnt 1 33,42 27,83 22,24 2 12,98 10,15 7,33 850C, 30mnt 1 7,84 8,61 9,38 2 9,25 9,90 10,54 1000C, 15mnt 1 13,88 16,00 18,12 2 11,70 11,83 11,95 1000C, 30mnt 1 12,98 12,08 11,18 2 12,47 11,05 9,64 74
Lampiran 7. c. Data inhibisi Acarbosa terhadap kerja enzim α-amilase Bacillus sp
- pH awal
Sampel Inhibisi (%) Rata-rata (%)
Acarbosa 98,76 98,76
98,76
- pH simulasi sistem pencernaan
Sampel Inhibisi (%) Rata-rata (%)
Acarbosa 65,21 65,21
65,21
73
Lampiran 8. Uji Anova daya inhibisi ekstrak rosella terhadap enzim α-amilase
Bacillus sp
a. pH awal
Univariate Analysis of Variance
Tests of Between-Subjects Effects
Source
Type III Sum of
Squares df Mean Square F Sig.
Corrected Model 70.461a 6 11.744 1.030 .477 Intercept 117401.149 1 117401.149 1.030E4 .000 Waktu .442 1 .442 .039 .850 Suhu 16.711 2 8.356 .733 .514 waktu * suhu 38.158 2 19.079 1.674 .255 Error 79.795 7 11.399 Total 127705.054 14 Corrected Total 150.256 13
b. pH saluran pencernaan
Univariate Analysis of Variance
Tests of Between-Subjects Effects
Source
Type III Sum of
Squares df Mean Square F Sig.
Corrected Model 5363.791a 6 893.965 33.466 .000 Intercept 10799.861 1 10799.861 404.298 .000 Waktu 638.502 1 638.502 23.903 .002 Suhu 320.246 2 160.123 5.994 .030 waktu * suhu 465.677 2 232.838 8.716 .013 Error 186.988 7 26.713 Total 13555.130 14 Corrected Total 5550.780 13
a. R Squared = .966 (Adjusted R Squared = .937)
Post Hoc Tests suhu Homogeneous Subsets Duncan suhu N Subset 1 2 3 100 4 12.7410 85 4 14.1228 70 4 24.3250 control 2 65.0000 Sig. .745 1.000 1.000
Means for groups in homogeneous subsets are displayed. Based on observed means.
75
Waktu Dependent Variable:amilase6.8
waktu Mean Std. Error
95% Confidence Interval Lower Bound Upper Bound
15 24.357 2.279 18.781 29.934
30 9.768 2.279 4.192 15.345
ANOVA amilase6.8
Sum of Squares Df Mean Square F Sig.
Between Groups 5363.791 6 893.965 33.466 .000
Within Groups 186.988 7 26.713
Total 5550.780 13
Post Hoc Tests Interaksi
Homogeneous Subsets
Duncan
Interaksi N
Subset for alpha = 0.05
1 2 3 70*30’ 2 8.4830 85*30’ 2 9.2545 100*30’ 2 11.5680 100*15’ 2 13.9140 85*15’ 2 18.9910 70*15’ 2 40.1670 Kontrol 2 65.0000 Sig. .099 1.000 1.000
Lampiran 9. Data inhibisi ekstrak rosella pH awal terhadap enzim α-amilase pankreas babi
pH awal
Kondisi Ekstraksi Ulangan Inhibisi (%) Rata-rata (%)
700C, 15menit 1 95,43 96,15
96,88
2 82,45 84,14
77
Lampiran 10.a. Data inhibisi ekstrak rosella pH awal terhadap enzim α-glukosidase
Kondisi Ekstraksi Ulangan Inhibisi (%) Rata-rata (%)
700C, 15mnt 1 91,77 90,59 89,41 2 95,29 96,47 97,65 700C, 30mnt 1 97,65 92,94 88,24 2 89,41 91,77 94,12 850C, 15mnt 1 88,24 88,24 88,24 2 88,24 87,65 87,06 850C, 30mnt 1 84,70 85,88 87,06 2 89,41 90,59 91,77 1000C, 15mnt 1 89,41 88,82 88,24 2 78,82 76,47 74,12 1000C, 30mnt 1 88,24 84,12 80,00 2 94,118 94,706 95,294 84
Lampiran 10.b. Data inhibisi ekstrak rosella pH pencernaan terhadap enzim α-glukosidase
Kondisi ekstraksi Ulangan Inhibisi (%) Rata-rata (%)
700C, 15mnt 1 37,28 36,95 36,63 2 42,29 43,83 45,37 700C, 30mnt 1 8,74 8,29 7,84 2 8,74 8,68 8,61 850C, 15mnt 1 33,42 27,83 22,24 2 12,98 10,15 7,33 850C, 30mnt 1 7,84 8,61 9,38 2 9,25 9,90 10,54 1000C, 15mnt 1 13,88 16,00 18,12 2 11,70 11,83 11,95 1000C, 30mnt 1 12,98 12,08 11,18 2 12,47 11,05 9,64
79
Lampiran 10.c. Data inhibisi acarbosa terhadap enzim α-glukosidase
- pH awal
Sampel Inhibisi (%) Rata-rata (%) Acarbosa 100,00 100,00
100,00
- pH saluran pencernaan
Sampel Inhibisi (%) Rata-rata (%) Acarbosa 100,00 100,00
100,00
Lampiran 11. Uji Anova daya inhibisi ekstrak rosella terhadap enzim α-glukosidase
a. pH awal
Univariate Analysis of Variance
Tests of Between-Subjects Effects Dependent Variable:gluksidase
Source
Type III Sum of
Squares df Mean Square F Sig.
Corrected Model 354.662a 6 59.110 2.561 .122 Intercept 107785.848 1 107785.848 4.669E3 .000 Waktu 11.533 1 11.533 .500 .503 Suhu 100.746 2 50.373 2.182 .183 waktu * suhu 35.696 2 17.848 .773 .497 Error 161.594 7 23.085 Total 115403.581 14 Corrected Total 516.256 13
81
Lampiran 12.a. Data inhibisi ekstrak rosella pH awal terhadap kerja enzim lipase
Kondisi Ekstraksi Ulangan Inhibisi (%) Rata-rata (%)
700C, 15mnt 1 89,66 88,95 88,24 2 91,31 90,21 89,11 700C, 30mnt 1 88,96 89,59 90,21 2 94,71 94,76 94,81 850C, 15mnt 1 89,16 89,39 89,61 2 96,33 95,73 95,13 850C, 30mnt 1 97,43 97,03 96,63 2 90,73 91,35 91,96 1000C, 15mnt 1 85,51 86,14 86,76 2 85,44 85,95 86,46 1000C, 30mnt 1 85,32 85,61 85,91 2 85,81 85,95 86,09 89 90
Lampiran 12.b. Data inhibisi ekstrak rosella pH pencernaan terhadap kerja enzim lipase
Kondisi Ekstraksi Ulangan Inhibisi (%) Rata-rata (%)
700C, 15menit 1 6,54 6,35 6,15 2 6,44 6,78 7,12 700C, 30menit 1 5,00 6,06 7,12 2 7,40 6,01 4,62 850C, 15menit 1 0,00 0,00 0,00 2 7,12 5,29 3,46 850C, 30menit 1 4,52 5,43 6,35 2 2,50 4,09 5,67 1000C, 15menit 1 50,58 49,33 48,078 2 32,98 32,98 32,98 1000C, 30menit 1 31,73 36,11 40,48 2 31,92 31,59 31,25 91
83
Lampiran 13. Uji Anova daya inhibisi ekstrak rosella terhadap enzim lipase a. pH awal
Univariate Analysis of Variance Tests of Between-Subjects Effects Dependent Variable:lipase
Source
Type III Sum of
Squares df Mean Square F Sig.
Corrected Model 124.723a 5 24.945 2.964 .109 Intercept 97316.135 1 97316.135 1.156E4 .000 Waktu 5.221 1 5.221 .620 .461 Suhu 115.288 2 57.644 6.850 .028 waktu * suhu 4.214 2 2.107 .250 .786 Error 50.494 6 8.416 Total 97491.351 12 Corrected Total 175.216 11
a. R Squared = .712 (Adjusted R Squared = .472)
Post Hoc Tests suhu Homogeneous Subsets Lipase Duncan suhu N Subset 1 2 100 4 85.9142 70 4 90.8748 90.8748 85 4 93.3722 Sig. .052 .269
Means for groups in homogeneous subsets are displayed.
Based on observed means.
Tests of Between-Subjects Effects Dependent Variable:lipase
Source
Type III Sum of
Squares df Mean Square F Sig.
Corrected Model 124.723a 5 24.945 2.964 .109 Intercept 97316.135 1 97316.135 1.156E4 .000 interaksi 124.723 5 24.945 2.964 .109 Error 50.494 6 8.416 Total 97491.351 12 Corrected Total 175.216 11
a. R Squared = .712 (Adjusted R Squared = .472)
Post Hoc Tests interaksi Homogeneous Subsets Duncan interaksi N Subset 1 2 100C30 2 85.7830 100C15 2 86.0455 70C15 2 89.5795 89.5795 70C30 2 92.1700 92.1700 85C15 2 92.5575 92.5575 85C30 2 94.1870 Sig. .070 .180
Means for groups in homogeneous subsets are displayed.
Based on observed means.
85
b. pH saluran pencernaan
Univariate Analysis of Variance Tests of Between-Subjects Effects Dependent Variable:lipase6.8
Source
Type III Sum of
Squares df Mean Square F Sig.
Corrected Model 3036.151a 5 607.230 25.230 .001 Intercept 2840.502 1 2840.502 118.023 .000 Waktu 3.247 1 3.247 .135 .726 Suhu 2959.155 2 1479.577 61.476 .000 waktu * suhu 73.749 2 36.875 1.532 .290 Error 144.405 6 24.067 Total 6021.057 12 Corrected Total 3180.555 11
a. R Squared = .955 (Adjusted R Squared = .917)
Post Hoc Tests suhu Homogeneous Subsets lipase6.8 Duncan Suhu N Subset 1 2 85 4 2.3800 70 4 6.2982 100 4 37.4778 Sig. .302 1.000
Means for groups in homogeneous subsets are displayed.
Based on observed means.
Source
Type III Sum of
Squares df Mean Square F Sig.
Corrected Model 3036.151a 5 607.230 25.230 .001 Intercept 2840.502 1 2840.502 118.023 .000 interaksi 3036.151 5 607.230 25.230 .001 Error 144.405 6 24.067 Total 6021.057 12 Corrected Total 3180.555 11
a. R Squared = .955 (Adjusted R Squared = .917)
Post Hoc Tests interaksi Homogeneous Subsets Duncan interaksi N Subset 1 2 85C15 2 .0000 85C30 2 4.7600 70C30 2 6.0340 70C15 2 6.5625 100C30 2 33.8015 100C15 2 41.1540 Sig. .248 .185
Means for groups in homogeneous subsets are displayed.
Based on observed means.
87
Lampiran 14. Penentuan konsentrasi ekstrak
[Ekstrak (M)] Ulangan Inhibisi (%) Rata-rata (%)
0,25 1 0,97 2 0,97 0,97 0,50 1 0,97 2 0,98 0,98 0,75 1 0,97 2 0,95 0,96 1 1 0,97 2 0,97 0,97 97
Lampiran 15. Kinetika inhibisi ekstrak rosela 1. Standar Maltosa [maltosa] (%) Absorbansi 0,00 0,235 0,20 0,406 0,40 0,557 0,60 0,699 0,80 0,868 1,00 0,998
Kurva standar maltosa
y = 0,7633x + 0,2455 R² = 0,9988 A b sor b an si Konsentrasi maltosa (%)
89
Lampiran 15 lanjutan 1. Standar (pati+ enzim)
[pati] Absorbansi [pati] bereaksi rataan Kecepatan rataan 1/[pati] 1/kecepatan rataan 1,000 0,8 0,73 0,07 1,000 13,77 1,250 0,98 0,96 0,09 0,800 10,39 1,500 1,145 1,18 0,12 0,667 8,49 1,750 1,269 1,34 0,13 0,571 7,46 2,000 1,456 1,59 0,16 0,500 6,31 2,250 1,646 1,84 0,18 0,444 5,45 2,500 1,853 2,11 0,21 0,400 4,75
2. Sampel (pati+enzim+ekstrak rosela)
[pati] Absorbansi [pati] bereaksi rataan Kecepatan rataan 1/[pati] 1/Kecepatan rataan 1,000 0,369 0,162 0,02 1,000 61,81 1,250 0,436 0,250 0,03 0,800 40,07 1,500 0,459 0,280 0,03 0,667 35,75 1,750 0,557 0,408 0,04 0,571 24,50 2,000 0,593 0,455 0,05 0,500 21,97 2,250 0,674 0,561 0,06 0,444 17,81 2,500 0,719 0,620 0,06 0,400 16,12
Lampiran 15. Lanjutan
1.Plot Lineweaver-Burk
Contoh perhitungan:
Plot Lineweaver-Burk inhibitor: 1/Vmaks = 15,65mM Vmaks = 1/ 15,65mM = 0,06 Km/ Vmaks = 74,78mM Km/ 0,06 = 74,78mM Km = 4,77mM y = 13,358x - 0,1936 R² = 0,9959 y = 56,013x - 2,7449 R² = 0,9541 1/V 1/ [S] non inhibitor inhibitor
