Lampiran 2.

Hasil Uji Kesesuian Sistem KCKT Shimadzu LC 20AD

Tabel V. Hasil Uji Kesesuaian Sistem KCKT Shimadzu LC 20AD

Gambar 5 . Kromatogram Hasil Uji Kesesuaian Sistem KCKT Shimadzu LC

20AD
Lampiran 2. (Lanjutan)

Tabel 1. Hasil Uji Kesesuaian Sistem Asam Retinoat

No. Title Ret. Time Area Plat Teori Faktor Ikutan

1 UKS 001 2.708 23135 3100.780 1.407
2 UKS 002 2.692 23402 2917.698 1.545
3 UKS 003 2.690 23479 2918.025 1.546
4 UKS 004 2.694 23641 2922.975 1.533
5 UKS 005 2.699 23650 2888.663 1.542
6 UKS 006 2.700 23752 2881.564 1.543
Rata-rata 2.697 23509.8 2938.284 1.519
SD 0.007 222.969 81.42841 0.055
%RSD 0.244 0.94841 2.771291 3.635

Tabel 2. Hasil Uji Kesesuaian Sistem Asam Retinoat

No. Title Ret. Time Area Plat Teori Faktor Ikutan

1 UKS 001 4.468 694411 3655.612 1.372
2 UKS 002 4.445 702524 3613.998 1.375
3 UKS 003 4.447 702979 3589.284 1.376
4 UKS 004 4.448 701707 3593.454 1.377
5 UKS 005 4.461 703416 3493.082 1.374
6 UKS 006 4.448 704093 3587.220 1.376
Rata-rata 4.453 701522 3588.775 1.375
SD 0.009 3575.6 53.4511079 0.002
%RSD 0.210 0.50969 1.48939702 0.130
Lampiran 3. Hasil Kurva Kalibrasi Asam Retinoat dan Hidrokuinon

Gambar 6. Gambar Kromatogram Kalibrasi Asam Retinoat dan Hidrokuinon

Tabel 3. Penimbangan Baku Asam Retinoat

Penimbangan Kadar Baku Susut Pengeringan Tertimbang
10.00 mg 100.24% 0.16% 10.0080 mg
Perhitungan :

Bobot Baku (Bb) = Baku Tertimbang x Kadar Baku x Susut Pengeringan

= 10,0 mg x 100,24% x (1-0,16%) = 10,008 mg

Tabel 4. Penimbangan Baku Hidrokuinon

Penimbangan Kadar Baku Susut Pengeringan Tertimbang
100.27 mg 100.35% 0.14% 100.4801 mg

Perhitungan :

Bobot Baku (Bb) = Baku Tertimbang x Kadar Baku x Susut Pengeringan

= 100,27 mg x 100,35% x (1-0,14%) = 100,48 mg

Lampiran 3. (Lanjutan)

Tabel 5. Hasil Kurva Kalibrasi Asam Retinoat

N
o Kons Baku Ret. Faktor Plat
Area
  Time ikutan teori
  (µg/mL)
1 11567 2.693 1.408 3069.49
2 1.001 11356 2.691 1.403 3054.56
3 11651 2.693 1.532 3079.19
11524.6 2.69233 1.4476 3067.74
Rata-rata
7 3 7 7
151.987 0.00115 0.0730
SD 12.4072
9 5 8
RSD 1.3188 0.0429 5.0480 0.4044
4 23135 2.708 1.407 3100.78
5 2.002 23402 2.692 1.545 2917.70
6 23749 2.690 1.546 2918.03
23428.6 2.69666 1.4993 2978.83
Rata-rata
7 7 3 4
307.867 0.00986 0.0799 105.608
SD
4 6 6 2
RSD 1.3141 0.3659 5.3333 3.5453
3056.59
7
34702 2.572 1.407 0
3088.11
8 3.002
34335 2.568 1.405 0
3098.42
9
34912 2.567 1.404 0
34649.6 1.4053
Rata-rata
7 2.569 3 3081.04
292.038 0.00264 0.0015 21.7927
SD
2 6 3 9
RSD 0.8428 0.1030 0.1087 0.7073
10 46270 2.762 1.485 3042.52
11 4.003 46235 2.765 1.484 3053.95
12 45992 2.771 1.487 3027.39
46165.6 1.4853 3041.28
Rata-rata 2.766
7 3 7
151.414 0.00458 0.0015 13.3228
SD
4 3 3 8
0.32798 0.16567 0.1028 0.43806
RSD
1 5 4 7
13 57837 2.776 1.484 3046.60
14 5.004 57645 2.777 1.418 3028.70
15 57991 2.781 1.482 3063.46
 57824.3 1.4613 3046.25
Rata-rata 2.778
3 3 3
173.347 0.00264 0.0375 17.3825
SD
4 6 4 9
0.29978 0.09523 2.5689 0.57062
RSD
3 9 6 2

Lampiran 3. (Lanjutan)

N
o Kons Faktor
Area Ret. Time Plat teori
  ikutan
  (µg/mL)
1 349986 4.493 1.308 3669.49
2 100.480 348555 4.491 1.303 3654.56
3 339972 4.493 1.332 3679.19
Rata-rata 346171 4.492 1.314 3667.747
SD 5415.962 0.001 0.016 12.407
RSD 1.565 0.026 1.180 0.338
4 694411 4.468 1.372 3655.61
5 200.960 702524 4.445 1.375 3589.28
6 702979 4.447 1.376 3589.28
699971.33
Rata-rata 4.4533 1.374 3611.393
3
SD 4820.761 0.0127 0.002 38.2945
RSD 0.689 0.2861 0.151 1.060
7 1049956 4.572 1.307 3656.590
8 301.440 1048867 4.568 1.050 3688.110
9 1047767 4.567 1.304 3698.420
1048863.3
Rata-rata
3 4.569 1.220 3681.040
SD 1094.505 0.003 0.148 21.793
RSD 0.104 0.058 12.089 0.592
10 1387772 4.462 1.385 3642.52
11 401.920 1387765 4.465 1.384 3653.95
12 1399222 4.471 1.387 3627.39
1391586.3
Rata-rata 4.466 1.385 3641.287
3
SD 6612.682 0.005 0.002 13.323
RSD 0.4752 0.103 0.110 0.3659
 13 1725342 4.476 1.384 3646.60
14 502.400 1736923 4.477 1.318 3628.70
15 1738888 4.481 1.382 3663.46
1733717.6
Rata-rata 4.478 1.361 3646.253
7
SD 7319.778 0.003 0.038 17.383
RSD 0.422 0.059 2.758 0.477
Tabel 6. Hasil Kurva Kalibrasi Hidrokuinon

Lampiran 3. (Lanjutan)

Kurva Kalibrasi Asam Retinoat

70000.00

60000.00 57824.33
f(x) = 11524.46 x + 117.7
50000.00 R² = 1
46165.67
40000.00
34649.67
30000.00 Linear ()
23428.67
20000.00

10000.00 11524.67

0.00
0.0000 1.0000 2.0000 3.0000 4.0000 5.0000 6.0000

Gambar 7. Gambar Kurva Kalibrasi Asam Retinoat

 2000000.00
Kurva Kalibrasi Hidrokuinon
1800000.00 1733717.67
1600000.00 f(x) = 3450.15 x + 4049.43
1400000.00 R² = 1 1391586.33
1200000.00
1000000.00 1048863.33
Linear ()
800000.00
699971.33
600000.00
400000.00 346171.00
200000.00
0.00
0.0000 100.0000 200.0000300.0000400.0000500.0000600.0000

Gambar 8. Gambar Kurva Kalibrasi Hidrokuinon

Lampiran 3 (Lanjutan)

Tabel 7. Hasil Persamaan Regresi Baku Asam Retinoat

Konsentras      
i Luas Area Rata-
No
Tiamfeniko Rata
.
l      
(mg/mL)        
 X y x.y x² y²
1 1.0008 11524.67 11533.84 1.001593 132817941.8
2 2.0016 23428.67 46894.64 4.006372 548902421.8
3 3.0024 34649.67 104031.76 9.014337 1200599400.1
16.02548
4
4.0032 46165.67 184809.69 7 2131268778.8
25.03982
5
5.0040 57824.33 289351.85 4 3343653525.4
Ʃ 15.01194 173593.00 636621.78 55.08761 7357242067.9
Ʃ² 225.35841        

Perhitungan :

n  xi. yi     xi    yi 
b =
 
n xi 2    xi 
2 = 11524,458

a =
 yi  b xi = 117,70
n

n  xi. yi     xi    yi 
n  xi     xi  . n  yi     yi  
= 0,9999
2 2 2 2

r =

Y = 11524,458 + 117,7x

Konsentrasi Luas Area Rata-

Tiamfenikol Rata
No.
(mg/mL)
Lampiran 3 (Lanjutan)
x y x.y x² y²
1
Tabel 100.480
8. Hasil 346171.00
Persamaan 34783288.28
Regresi Baku 10096.245608
Hidrokuinon 119834361241.0
2 200.960 699971.33 140666345.09 40384.982432 489959867488.4
3 301.440 1048863.33 316169601.32 90866.210473 1100114292011.1
4 401.920 1391586.33 559306800.34 161539.929729 1936512523120.1
5 502.400 1733717.67 871020411.75 252406.140201 3005776947712.1
Ʃ 1507.201 5220309.67 1921946446.78 555293.50844 6652197991572.8
Ʃ² 2271655.262
Perhitungan :

n  xi. yi     xi    yi 
b =
 
n xi 2    xi 
2 = 4049,433

a =
 yi  b xi = 3450,145
n

n  xi. yi     xi    yi 
n  xi     xi  . n  yi     yi  
= 0,9999
2 2 2 2

r =

Y = 3450,145 + 4049,433

Lampiran 4. Hasil Uji Penetapan Kadar

Gambar 9. Gambar Kromatogram Penetapan Kadar Sampel A

Lampiran 4. (Lanjutan)
Lampiran 4. (Lanjutan)
Gambar 10. Gambar Kromatogram Penetapan Kadar Sampel B
Lampiran 4. (Lanjutan)
Lampiran 4. (Lanjutan)
Gambar 10. Gambar Kromatogram Penetapan Kadar Sampel C
Lampiran 4. (Lanjutan)
Lampiran 4. (Lanjutan)
Gambar 11. Gambar Kromatogram Penetapan Kadar Sampel D
Lampiran 4. (Lanjutan)
Gambar 11. Gambar Kromatogram Penetapan Kadar Sampel D
Lampiran 4. (Lanjutan)
Gambar 12. Gambar Kromatogram Penetapan Kadar Sampel E
Lampiran 4. (Lanjutan)
Gambar 12. Gambar Kromatogram Penetapan Kadar Sampel E
Lampiran 4. (Lanjutan)
Tabel 9. Hasil Uji Penetapan Kadar Asam Retinoat

No
Sampel Area Konsentrasi / Kadar
 
          (µg/mL) (µg) (mg) (%)
1 Sampel A 1 22858   1.973 197.322 0.197 0.197
    2 23165   2.000 199.986 0.200 0.198
    3 24550   2.120 212.004 0.212 0.210
Rata-rata 2.031 203.104 0.203 0.202
SD 0.0782 7.8218 0.0078 0.0074
RSD 3.8511 3.8511 3.8511 3.6530
2 Sampel B 1 0   - - - -
    2 0   - - - -
    3 0   - - - -
Rata-rata - - - -
SD - - - -
RSD - - - -
3 Sampel C 1 57368   4.968 496.772 0.497 0.494
    2 59414   5.145 514.526 0.515 0.514
    3 59616   5.163 516.279 0.516 0.513
Rata-rata 5.092 509.192 0.509 0.507
SD 0.1079 10.7916 0.0108 0.0114
RSD 2.1194 2.1194 2.1194 2.2570
4 Sampel D 1 0   - - - -
    2 0   - - - -
    3 0   - - - -
Rata-rata - - - -
SD - - - -
RSD - - - -
5 Sampel E 1 0   - - - -
    2 0   - - - -
    3 0   - - - -
Rata-rata - - - -
SD - - - -
RSD - - - -

Contoh Perhitungan Kadar sampel Asam Retinoat :

Hasil Persamaan Regresi Kurva Klibrasi Asam Retinoat

Y = a + bx
Y = 117,7 + 11524,458x
Kadar Sampel A (1) :
X (µg/mL) = y-b
a
= 22858 -11524
117.700
= 1.973
Csp (x) x Volume
Kadar (µg) = Pengenceran
= 1.973 x 100 mL
= 197.322 µg/100 mL
= 0.19732 mg/100 mL
= 0.19732 %
Kadar Sampel A (2) :
X (µg/mL) = y-b
a
= 23165 -11524
117.700
= 2.000
Csp (x) x Volume
Kadar (µg) = Pengenceran
= 1.9999 x 100 mL
= 199.986 µg/100 mL
= 0.19999 mg/100 mL
= 0.19999 %
Kadar Sampel A (3) :
X (µg/mL) = y-b
a
= 24550 -11524
117.700
= 2.120
Csp (x) x Volume
Kadar (µg) = Pengenceran
= 2.120 x 100 mL
= 212.004 µg/100 mL
= 0.212 mg/100 mL
= 0.212 %
Rata-rata Kadar (1+2+3)
= 0.197 + 0.19999 0.212
= 0.202 %
Lampiran 4. (Lanjutan)

Tabel 9. Hasil Uji Penetapan Kadar Hidrokuinon

N
o Sampel Area Konsentrasi / Kadar
 
          (µg/mL) (µg) (mg/100 mL) (%)
Sampel 19.19
1 A 1 666218   191.925 19192.485 19.192 2
19.17
    2 665604   191.747 19174.689 19.175 5
19.61
    3 680719   196.128 19612.786 19.613 3
19.32
Rata-rata 193.267 19326.653 19.327
7
SD 2.480 247.958 0.248 0.248
RSD 1.283 1.283 1.283 1.283
Sampel 31.35
2 B 1 1085678   313.502 31350.235 31.350 0
31.27
    2 1082938   312.708 31270.818 31.271 1
30.91
    3 1070770   309.181 30918.137 30.918 8
31.18
Rata-rata 311.797 31179.730 31.180
0
SD 2.300 230.000 0.230 0.230
RSD 0.738 0.738 0.738 0.738
Sampel 45.31
3 C 1 1567449   453.140 45314.025 45.314 4
43.80
    2 1515206   437.998 43799.798 43.800 0
43.84
    3 1516682   438.426 43842.579 43.843 3
44.31
Rata-rata 443.188 44318.801 44.319
9
SD 8.622 862.155 0.862 0.862
RSD 1.945 1.945 1.945 1.945
Sampel 41.57
4 D 1 1438520   415.771 41577.109 41.577 7
41.09
    2 1421840   410.937 41093.651 41.094 4
40.76
    3 1410544   407.662 40766.245 40.766 6
41.14
Rata-rata 411.457 41145.668 41.146
6
SD 4.079 407.927 0.408 0.408
RSD 0.991 0.991 0.991 0.991
 Sampel 39.93
5 E 1 1381993   399.387 39938.714 39.939 9
41.22
    2 1426217   412.205 41220.516 41.221 1
40.81
    3 1412254   408.158 40815.808 40.816 6
40.65
Rata-rata 406.583 40658.346 40.658
8
SD 6.552 655.247 0.655 0.655
RSD 1.612 1.612 1.612 1.612

Contoh Perhitungan Kadar sampel Hidrokuinon :

Hasil Persamaan Regresi Kurva Klibrasi Hidrokuinon

Y = a + bx
Y = 3450,145 + 4049,433x
Kadar Sampel A (1) :
X (µg/mL) = y-b
a
= 666218 -3450.145
4049.433
= 191.925
Kadar
(µg) = Csp (x) x Volume Pengenceran
= 191.925 x 100 mL
= 19192.485 µg/100 mL
= 19.192485 mg/100 mL
= 19.192485 %
Kadar Sampel A (2) :
X (µg/mL) = y-b
a
= 665604 -3450.145
4049.433
= 191.747
Kadar
(µg) = Csp (x) x Volume Pengenceran
= 191.747 x 100 mL
= 19174.689 µg/100 mL
= 19.174689 mg/100 mL
= 19.174689 %
Kadar Sampel A (3) :
X (µg/mL) = y-b
a
= 680719 -3450.145
4049.433
= 196.128
Kadar
(µg) = Csp (x) x Volume Pengenceran
= 196.128 x 100 mL
= 19612.786 µg/100 mL
= 19.612786 mg/100 mL
= 19.612786 %
Rata-rata Kadar
(1+2+3)
19.19 19.17
= 2 + 5 + 19.613
19.32
= 7 %

Lampiran 5. Alat dan Instrumen yang digunakan

Gambar 19. Gambar KCKT Shimadzu LC-20AD

 Gambar 20. Gambar Kolom X-Terra RP18 5µm ukuran 4,6 x 250 mm
by Waters (18600496)

Gambar 20. Gambar Timbangan analitik digital Radwag XA 82/220/2X

Lampiran 12. (Lanjutan

Gambar 21. Gambar Timbangan Micro Balance RADWAG MYA2

 Gambar 22. Gambar Sonikator Branson

Gambar 23. Gambar Pompa Vakum Sartorius