Nama : Lieneilson Firdaus Nilai :

Kelas : 13-1
NIS : 16.62.08392
No. Meja :4
Tanggal : 18 Juli 2019

JUDUL PENETAPAN:
PENETAPAN KADAR THIAMIN PADA TABLET VITAMIN B1 DENGAN HPLC
I. Prinsip Analisis (20) :
Vitamin B1 dapat dipisahkan dan dianalisis menggunakan teknik HPLC. Vitamin B1
merupakan salah satu vitamin yang dapat terlarut di dalam air , sehingga digunakan fase
gerak berupa pelarut buffer fosfat dan methanol yang polar dan digunakan kolom yang
bersifat cenderung nonpolar. Vitamin dapat dideteksi melalui detector VWD dengan panjang
gelombang 254 nm.

II. Persiapan Sampel (20) :

a. Identifikasi sampel
Kode sampel : 013.4-2/LABTUK/HPLC/2017
Tgl. Penerimaan : 22 November 2019
Sifat fisik sampel
- Wujud padat/cair/gas
- Warna : Putih
- Bau : Normal

b. Persiapan sampel
1) Timbang 5 tablet vitamin B1 lalu rata-ratakan bobotnya. Ambil 2 tablet lalu haluskan.

2) Ditimbang + 0,2000 gram contoh vitamin B1 lalu dimasukkan ke dalam labu ukur 100 mL,
lalu ditambahkan buffer fosfat sampai tanda tera.

3) Setelah itu dikocok selama 5 menit, lalu dibiarkan mengendap dan disaring dengan kertas
saring Whatman 41. Pipet 5 mL filtrat ke dalam labu ukur 50 mL, lalu diimpitkan sampai
tanda tera, dikocok lalu disaring dengan kertas saring millipore. Filtratnya siap untuk
diinjeksikan.
 PERSIAPAN STANDARD

1) Pembuatan Larutan Buffer Fosfat 0,04M

Ditimbang 10,8872 ±0,0005 gram KH2PO4, dimasukkan ke dalam labu ukur 2000 mL,
diencerkan dengan akuabides, diimpitkan pada skala tera, dihomogenkan. Ditempatkan
pada botol dan diberi label yang sesuai.

2) Pembuatan larutan induk thiamin 1000 ppm

Timbang standard thiamin-HCl(pa) sebanyak 0,1000 gram lalu encerkan dalam labu ukur
100 mL dengan buffer fosfat. Kocok sampai homogen.

3) Pembuatan larutan kerja thiamin 100 ppm

Dipipet 10 mL larutan induk 1000 ppm lalu diencerkan dengan buffer fosfat sampai volume
100 mL.

4) Larutan Deret Standar Tiamina: 0 - 50 ppm. Diturunkan dari buret standar induk tiamina
100 ppm sejumlah 0 mL; 2,5 mL; 5 mL; 7,5 mL; 10 mL; 15 mL; dan 25 mL ke dalam labu
ukur 50 mL, diencerkan dan diimpitkan hingga 50 mL buffer fosfat, dihomogenkan. Lalu
disaring dengan kertas saring millipore dan filtratnya siap untuk diinjeksikan.

III. Data Hasil Pengamatan (15):

Data Penimbangan :
Bobot Kaca Arloji + 5 tablet : 22.7886 g
Bobot Kaca Arloji Kosong : 21.7791 g
Bobot 5 tablet : 1.0095 g

Bobot Kaca Arloji + Thiamin : 28.2080 g

Bobot Kaca Arloji Kosong : 28.0063 g
Bobot Thiamin-HCl (p.a) : 0.1010 g

Bobot Kaca Arloji + Sampel : 22.6384 g

Bobot Kaca Arloji Kosong : 22.4347 g
Bobot Sampel (duplo) : 0.2010 g

Bobot Kaca Arloji + Sampel : 28.2080 g

Bobot Kaca Arloji Kosong : 28.0066 g
Bobot Sampel (simplo) : 0.2014 g

r = 0.997
r2 = 0.995
Intercept = 5.164
Slope = 23.0588
No Nama Waktu retensi Area Keterangan
1 Standard thiamin
0 ppm 0 0 Tidak Ada
5 ppm 1.747 124.19849 Tidak Ada
10 ppm 1,722 212.93108 Tidak Ada
15 ppm 1.646 403.65601 Tidak Ada
20 ppm 1.701 456.92447 Tidak Ada
30 ppm 1,746 669.12628 Tidak Ada
50 ppm 1.758 1166.96216 Tidak Ada
2 Sampel Vitamin B1
Simplo 1.711 603.53076 Tidak Ada
Duplo 1.719 598.85256 Tidak Ada
IV. Pengolahan Data Hasil pengukuran (25):
𝟏.𝟎𝟎𝟗𝟓 𝒈
Bobot Rata-rata Tablet : = 𝟎. 𝟐𝟎𝟏𝟗 𝒈 = 201.9 mg
𝟓
𝟔𝟎𝟑.𝟓𝟑𝟎𝟕𝟖−𝟓.𝟏𝟔𝟒
( )𝒙𝟏𝟎𝒙𝟎.𝟏
𝟐𝟑.𝟎𝟓𝟖𝟖
mg/tablet(s) = 𝟐𝟎𝟏.𝟒
𝒙𝟐𝟎𝟏. 𝟗 = 𝟐𝟔. 𝟎𝟏 𝒎𝒈/𝒕𝒂𝒃𝒍𝒆𝒕
𝟓𝟗𝟖.𝟖𝟓𝟐𝟓𝟔−𝟓.𝟏𝟔𝟒
( )𝒙𝟏𝟎𝒙𝟎.𝟏
𝟐𝟑.𝟎𝟓𝟖𝟖
mg/tablet(d) = 𝒙𝟐𝟎𝟏. 𝟗 = 𝟐𝟓. 𝟖𝟔 𝒎𝒈/𝒕𝒂𝒃𝒍𝒆𝒕
𝟐𝟎𝟏.𝟎

𝟐𝟔.𝟎𝟏−𝟐𝟓.𝟖𝟔
RPD = | 𝟐𝟔.𝟎𝟔+𝟐𝟓.𝟖𝟔 | 𝒙𝟏𝟎𝟎% = 𝟎. 𝟓𝟖%
𝟐

𝟐𝟔.𝟎𝟔+𝟐𝟓.𝟖𝟔
Rata-rata mg/tablet = = 𝟐𝟓. 𝟗𝟑𝟓 𝒎𝒈/𝒕𝒂𝒃𝒍𝒆𝒕
𝟐
(𝟐𝟓.𝟗𝟑𝟓−𝟐𝟓)
Simpangan dari Label Klaim = 𝒙𝟏𝟎𝟎% = 𝟑. 𝟕𝟒%
𝟐𝟓

V. Kesimpulan (20) :
Dari penetapan kadar Thiamin dalam Vitamin B1 dengan HPLC didapat jumlah rata-
rata Thiamin dalam sampel sebanyak 25.935 mg/tablet dengan RPD 0.57%. Label klaim
sampel 25 mg/ tablet sehingga simpangan dari label klaim sebesar 3.74%.

Bogor, 18 Juli 2019

Guru Praktik Siswa

(...................................) (Lieneilson Firdaus)

Data File C:\HPCHEM\1\DATA\MNTTEST\DEFAULT.D Sample Name: Standar 5 ppm

=====================================================================
Injection Date : 7/18/2019 11:45:28 AM
Sample Name : Standar 5 ppm Location : Vial 1
Acq. Operator : Kelompok Karbon
Acq. Instrument : Instrument 1
Method : C:\HPCHEM\1\METHODS\THIAMINE.M
Last changed : 7/18/2019 11:39:05 AM by besi
(modified after loading)
VWD1 A, Wavelength=254 nm (MNTTEST\DEFAULT.D)

1.747
mAU

-1

-2

-3
0 1 2 3 4 min

=====================================================================
Area Percent Report
=====================================================================

Sorted By : Signal
Multiplier : 1.0000
Dilution : 1.0000
Sample Amount : 1.00000 [ng/ul] (not used in calc.)
Use Multiplier & Dilution Factor with ISTDs

Signal 1: VWD1 A, Wavelength=254 nm

Peak RetTime Type Width Area Height Area

# [min] [min] mAU *s [mAU ] %
----|-------|----|-------|----------|----------|--------|
1 1.747 VV 0.2232 124.19849 6.94409 100.0000

Totals : 124.19849 6.94409

Results obtained with enhanced integrator!

=====================================================================
*** End of Report ***

Instrument 1 7/18/2019 11:50:36 AM Kelompok Karbon Page 1 of 1

Data File C:\HPCHEM\1\DATA\MNTTEST\DEFAULT.D Sample Name: Standar 10 ppm

=====================================================================
Injection Date : 7/18/2019 11:54:57 AM
Sample Name : Standar 10 ppm Location : Vial 1
Acq. Operator : Kelompok Karbon
Acq. Instrument : Instrument 1
Method : C:\HPCHEM\1\METHODS\THIAMINE.M
Last changed : 7/18/2019 11:39:05 AM by besi
(modified after loading)
VWD1 A, Wavelength=254 nm (MNTTEST\DEFAULT.D)

1.722
mAU

10

0 1 2 3 4 min

=====================================================================
Area Percent Report
=====================================================================

Sorted By : Signal
Multiplier : 1.0000
Dilution : 1.0000
Sample Amount : 1.00000 [ng/ul] (not used in calc.)
Use Multiplier & Dilution Factor with ISTDs

Signal 1: VWD1 A, Wavelength=254 nm

Peak RetTime Type Width Area Height Area

# [min] [min] mAU *s [mAU ] %
----|-------|----|-------|----------|----------|--------|
1 1.722 VB 0.2403 212.93106 11.07683 100.0000

Totals : 212.93106 11.07683

Results obtained with enhanced integrator!

=====================================================================
*** End of Report ***

Instrument 1 7/18/2019 12:00:04 PM Kelompok Karbon Page 1 of 1

Data File C:\HPCHEM\1\DATA\MNTTEST\DEFAULT.D Sample Name: Standar 15 ppm

=====================================================================
Injection Date : 7/18/2019 12:02:41 PM
Sample Name : Standar 15 ppm Location : Vial 1
Acq. Operator : Kelompok Karbon
Acq. Instrument : Instrument 1
Method : C:\HPCHEM\1\METHODS\THIAMINE.M
Last changed : 7/18/2019 11:39:05 AM by besi
(modified after loading)
VWD1 A, Wavelength=254 nm (MNTTEST\DEFAULT.D)

1.646
mAU
14

12

10

6
1.447

0 1 2 3 4 min

=====================================================================
Area Percent Report
=====================================================================

Sorted By : Signal
Multiplier : 1.0000
Dilution : 1.0000
Sample Amount : 1.00000 [ng/ul] (not used in calc.)
Use Multiplier & Dilution Factor with ISTDs

Signal 1: VWD1 A, Wavelength=254 nm

Peak RetTime Type Width Area Height Area

# [min] [min] mAU *s [mAU ] %
----|-------|----|-------|----------|----------|--------|
1 1.447 BV 0.1728 51.18244 3.88117 11.2529
2 1.646 VB 0.3531 403.65601 13.90882 88.7471

Totals : 454.83844 17.78999

Results obtained with enhanced integrator!

=====================================================================
*** End of Report ***

Instrument 1 7/18/2019 12:07:49 PM Kelompok Karbon Page 1 of 1

Data File C:\HPCHEM\1\DATA\MNTTEST\DEFAULT.D Sample Name: Standar 20 ppm

=====================================================================
Injection Date : 7/18/2019 12:12:38 PM
Sample Name : Standar 20 ppm Location : Vial 1
Acq. Operator : Kelompok Karbon
Acq. Instrument : Instrument 1
Method : C:\HPCHEM\1\METHODS\THIAMINE.M
Last changed : 7/18/2019 11:39:05 AM by besi
(modified after loading)
VWD1 A, Wavelength=254 nm (MNTTEST\DEFAULT.D)

1.701
mAU

20

15

10

0 1 2 3 4 min

=====================================================================
Area Percent Report
=====================================================================

Sorted By : Signal
Multiplier : 1.0000
Dilution : 1.0000
Sample Amount : 1.00000 [ng/ul] (not used in calc.)
Use Multiplier & Dilution Factor with ISTDs

Signal 1: VWD1 A, Wavelength=254 nm

Peak RetTime Type Width Area Height Area

# [min] [min] mAU *s [mAU ] %
----|-------|----|-------|----------|----------|--------|
1 1.701 VB 0.2328 456.92477 24.26321 100.0000

Totals : 456.92477 24.26321

Results obtained with enhanced integrator!

=====================================================================
*** End of Report ***

Instrument 1 7/18/2019 12:17:46 PM Kelompok Karbon Page 1 of 1

Data File C:\HPCHEM\1\DATA\MNTTEST\DEFAULT.D Sample Name: Standar 30 ppm

=====================================================================
Injection Date : 7/18/2019 12:19:30 PM
Sample Name : Standar 30 ppm Location : Vial 1
Acq. Operator : Kelompok Karbon
Acq. Instrument : Instrument 1
Method : C:\HPCHEM\1\METHODS\THIAMINE.M
Last changed : 7/18/2019 11:39:05 AM by besi
(modified after loading)
VWD1 A, Wavelength=254 nm (MNTTEST\DEFAULT.D)

1.746
mAU

35

30

25

20

15

10

0 1 2 3 4 min

=====================================================================
Area Percent Report
=====================================================================

Sorted By : Signal
Multiplier : 1.0000
Dilution : 1.0000
Sample Amount : 1.00000 [ng/ul] (not used in calc.)
Use Multiplier & Dilution Factor with ISTDs

Signal 1: VWD1 A, Wavelength=254 nm

Peak RetTime Type Width Area Height Area

# [min] [min] mAU *s [mAU ] %
----|-------|----|-------|----------|----------|--------|
1 1.746 VB 0.2122 669.12628 39.48990 100.0000

Totals : 669.12628 39.48990

Results obtained with enhanced integrator!

=====================================================================
*** End of Report ***

Instrument 1 7/18/2019 12:24:38 PM Kelompok Karbon Page 1 of 1

Data File C:\HPCHEM\1\DATA\MNTTEST\DEFAULT.D Sample Name: Standar 50 ppm

=====================================================================
Injection Date : 7/18/2019 12:26:40 PM
Sample Name : Standar 50 ppm Location : Vial 1
Acq. Operator : Kelompok Karbon
Acq. Instrument : Instrument 1
Method : C:\HPCHEM\1\METHODS\THIAMINE.M
Last changed : 7/18/2019 11:39:05 AM by besi
(modified after loading)
VWD1 A, Wavelength=254 nm (MNTTEST\DEFAULT.D)

1.758
mAU

60

50

40

30

20

10

0 1 2 3 4 min

=====================================================================
Area Percent Report
=====================================================================

Sorted By : Signal
Multiplier : 1.0000
Dilution : 1.0000
Sample Amount : 1.00000 [ng/ul] (not used in calc.)
Use Multiplier & Dilution Factor with ISTDs

Signal 1: VWD1 A, Wavelength=254 nm

Peak RetTime Type Width Area Height Area

# [min] [min] mAU *s [mAU ] %
----|-------|----|-------|----------|----------|--------|
1 1.758 VB 0.2198 1166.96216 66.81720 100.0000

Totals : 1166.96216 66.81720

Results obtained with enhanced integrator!

=====================================================================
*** End of Report ***

Instrument 1 7/18/2019 12:31:48 PM Kelompok Karbon Page 1 of 1

Data File C:\HPCHEM\1\DATA\MNTTEST\DEFAULT.D Sample Name: simplo

=====================================================================
Injection Date : 7/18/2019 12:33:15 PM
Sample Name : simplo Location : Vial 1
Acq. Operator : Kelompok Karbon
Acq. Instrument : Instrument 1
Method : C:\HPCHEM\1\METHODS\THIAMINE.M
Last changed : 7/18/2019 11:39:05 AM by besi
(modified after loading)
VWD1 A, Wavelength=254 nm (MNTTEST\DEFAULT.D)

1.711
mAU

30

25

20

15

10

0 1 2 3 4 min

=====================================================================
Area Percent Report
=====================================================================

Sorted By : Signal
Multiplier : 1.0000
Dilution : 1.0000
Sample Amount : 1.00000 [ng/ul] (not used in calc.)
Use Multiplier & Dilution Factor with ISTDs

Signal 1: VWD1 A, Wavelength=254 nm

Peak RetTime Type Width Area Height Area

# [min] [min] mAU *s [mAU ] %
----|-------|----|-------|----------|----------|--------|
1 1.711 VP 0.2238 603.53076 33.89360 100.0000

Totals : 603.53076 33.89360

Results obtained with enhanced integrator!

=====================================================================
*** End of Report ***

Instrument 1 7/18/2019 12:38:23 PM Kelompok Karbon Page 1 of 1

Data File C:\HPCHEM\1\DATA\MNTTEST\DEFAULT.D Sample Name: duplo

=====================================================================
Injection Date : 7/18/2019 12:39:41 PM
Sample Name : duplo Location : Vial 1
Acq. Operator : Kelompok Karbon
Acq. Instrument : Instrument 1
Method : C:\HPCHEM\1\METHODS\THIAMINE.M
Last changed : 7/18/2019 11:39:05 AM by besi
(modified after loading)
VWD1 A, Wavelength=254 nm (MNTTEST\DEFAULT.D)

1.719
mAU
35

30

25

20

15

10

0 1 2 3 4 min

=====================================================================
Area Percent Report
=====================================================================

Sorted By : Signal
Multiplier : 1.0000
Dilution : 1.0000
Sample Amount : 1.00000 [ng/ul] (not used in calc.)
Use Multiplier & Dilution Factor with ISTDs

Signal 1: VWD1 A, Wavelength=254 nm

Peak RetTime Type Width Area Height Area

# [min] [min] mAU *s [mAU ] %
----|-------|----|-------|----------|----------|--------|
1 1.719 VB 0.2119 598.85254 35.40504 100.0000

Totals : 598.85254 35.40504

Results obtained with enhanced integrator!

=====================================================================
*** End of Report ***

Instrument 1 7/18/2019 12:44:49 PM Kelompok Karbon Page 1 of 1