Vitamins

VITAMIN C (ASCORBIC ACID)

B. IODINE TITRATION METHOD

I l

+ I2

Kadar Vitamin C = ml iodine x N iodine X BM Vitamin c x 100%

2 x berat sampel (gr) x 1000
 Prinsip analisis
 Oksidasi vitamin C oleh iodin membentuk
dehidroasamaskorbat :
 C6H8O6(aq) + I2(aq)  C6H6O6(aq) + 2 I- (aq) + 2 H+
Vitamin-C (ascorbic acid) (dehydroascorbic acid)

 Due to this reaction the iodine formed is immediately

reduced to iodide as long as there is any ascorbic acid
present. Once all the ascorbic acid has been
oxidised, the excess iodine is free to react with the
starch indicator, forming the blue-black starch-iodine
complex. This is the endpoint of the titration
Titration of a colour-free Vitamin C tablet Analysis of Vitamin C in a commercial fruit juice

Analysis of Vitamin C in freshly Analysis of Vitamin C in red capsicum (pepper

squeezed orange juice
 B. 2,6 D TITRATION METHOD

In acidic solutions DCIP is red, but if ascorbic acid is present, it will be reduced
to a colorless substance. The solution will remain colorless as more DCIP is
added until all of the ascorbic acid has reacted. As soon as the next drop of
DCIP solution is added at the solution will be light red, due to the excess DCIP
and the end point of the titration has been reached.
 Need standardization of the 2, 6-Dichloroindophenol solution once a day
with a solution of ascorbic acid with a known concentration
 Prinsip analisis
 Ox: vitamin Cred → vitamin Cox + 2 e− +
2 H+ (1)
 Red: DCPox + 2 e− + 2 H+ → DCPred (2)
 Overall: vitamin Cred + DCPox → vitamin
Cox + DCPred (3)
 Calculation for Reduced Vitamin C

F V
mg Ascorbic Acid/g = (A-B) x x
E X

A= ml of 2,6-dichloroindophenol for sample titration

B= ml of 2,6-dichloroindophenol for blank
F= mg of ascorbic acid equivalent to 1 ml of indophenol standard solution
V= Initial assay solution volume
E= Number of grams
X= Volume sample aliquot titrated
 Contoh soal
 Dilakukan analisis kadar vitamin C dengan titrasi iodometri. 20
gram sampel buah mangga dengan kadar air 25% diekstraksi
vitamin Cnya menggunakan aquadest hingga volume totalnya
250 ml, disaring hingga diperoleh sari buah mangga 220 ml.
Diambil 20 ml sari buah dimasukkan dalam erlenmeyer
ditambah 30 aquadest, ditambah 2 ml indikator amilum dan
dititrasi dengan 0,0097 N larutan Iodin yang telah
distandardisasi. Tentukan kadar vitamin C buah mangga
tersebut dalam wb dan db, jika larutan iodin yang diperlukan
untuk mengadisi ikatan rangkap vitamin C (BM 176.12)
sampel adalah 3.7 ml
 Jawab: kadar vit C (%wb) = 0.174 % , = 1740 mg/kg bb
 Kadar Vitamin C (%db) = 0.232 % = 2320 mg/kg bk
 Contoh soal
 Kadar Vitamin C buah salak ditentukan dengan metode 2.6D.
25 gram buah salak dengan kadar air 18% dihancurkan
dengan penambahan aquadest 150 ml. Ekstrak buah salak
yang dihasilkan ditambah aquadest hingga valume totalnya
200 ml, disaring dan diperoleh sari buah salak sebanyak 175
ml. 20 ml sari buah salak, ditambah 30 ml aquadest, dititrasi
dengan larutan 2.6D. Warna merah terbentuk saat 5.4 ml
larutan 2.6D ditambahkan. Jika tiap ml larutan 2.6D
equivalen dengan 0.25 mg vitamin C. Tentukan kadar
vitamin C buah salak tersebut dalam wb dan db.
 Jawab:
 kadar vitC = 0.54 mg/g bb = 540 mg/ kg bb = 0.054% wb
 Kadar Vit C = 0.659 mg/g bk = 659 mg/ kg bk = 0.066 % db
 Vitamin A (Retinol)

CH3 CH3
H3C CH3
OH

Retinol
CH3
b - Carotene and Retinol
CH 3
CH 3 CH 3 CH 3
H3 C

CH 3
CH 3 CH 3 CH 3
CH 3

Oxidation

CH 3 CH 3 O
H3 C CH 3
C H

CH 3 Retainal
- 2H

CH 3 CH 3
H3 C CH 3
CH 2 OH

CH 3 Retinol (Vitamin A)
Vitamin A Activity Determnination in Foods

Vitamin A Activity = b-Carotene + Retinol

Vitamin A and b - Carotene Determination
Food
KOH (Alcoholic)
Saponification

3 hrs. at room temperature

Ether for Extraction
Extract (retinol and b-carotene)

HPLC
 Vitamin A

 Column: Nova-Pack C18; 150 x 3.9mm

 Eluent : 95:5 Methanol: CH3CN
 Detection wavelength: 285 nm