Analisis karbohidrat

Bagian 1

overview
Pendahuluan Preparasi Sampel Mono- dan Oligosakarida Polisakarida Dietray Fiber

Komponen Alami:
Vitamin & Mineral

Komponen Tambahan:
Emulsifier Antioxidant

Lipida Asam Protein Pigmen Carbohydrate Antioksidan Water Citarasa Enzim

Preservative Thickener Sweetener Etc.

MUTU & KEAMANAN PANGAN

Pendahuluan

Karbohidrat merupakan komponen dalam makanan yang merupakan sumber energi utama bagi organisme hidup. senyawa organik yang terdiri dari bahan ekstrak tanpa nitrogen (BETN ) dan serat kasar, unsurnya terdiri dari karbon (C), hidrogen (H) dan oksigen (O). senyawa polihidroksi aldehid atau polihhidroksi keton (CnH2nOn). Berdasarkan jumlah monomer penyusunnya, dapat dikelompokkan mjd mono-, di-, oli-, dan polisakarida.

Pendahuluan

Pendahuluan
Monosakarida mungkin dapat bergabung satu sama lain, dengan mengambil satu molekul air untuk tiap gabungan, menghasilkan, di-, tri-, atau polisakarida yang mengandung 2,3 atau lebih monosakarida. KH dibagi menjadi 2 golongan gula (monotetrasakarida) dan non-gula (homopolisakarida, heteropolisakarida). HOMOPOLISAKARIDE

Pentosan Heksosan

HETEROPOLISAKARIDE
Hemiselulose Gummi Musilage Zat Peptik Mukopolisakaride dari hewan

CARBOHYDRATE
Cx(H2O)y

Mono saccharides Hexose (C-6)

a. b. c. d. D-Glucose D-Galactose D-Mannose D-Fructose

Di saccharides
Sucrose (a+d) Lactose (a+b) Maltose (a+a) -> a Cellobiose (a+a) ->b

Oligo saccharides
Rafinose (b+a+d) Stachyose (b+b+a+d) Verbacose

Pentose (C-5)
1. Xylose 2. Arabinose 3. Ribose

POLY SACCHARIDES
Cellulose (-> a) Starch (-> a) Chitin Agar (->b) Carrageenan (->b) Alginat

Pendahuluan
Humans can digest only sucrose, lactose, maltooligosaccharides/ maltodextrins, and starch. All are digested with enzymes found in the small intestine. Hanya amilum (jenis polisakarida) yg dpat dicerna manusia. Nondigestible polysaccharides can be divided into soluble and insoluble classes. Along with lignin and other nondigestible, nonabsorbed substances, they make up dietary fiber (sbg komponen tambahan dlm produk pangan)

POLYSACCHARIDES

STARCH Daratan: Cellulose Pectin Gum Lainnya

NON-STARCH POLYSACCHARIDES Perairan:

Alginate
Carrageenan

Agar
Furcellaran Chitosan Lainnya

Struktur Kimia
Gugus Aldehid (CHO) = gugus reduksi H
1

Gugus Karbonil (CO)

C C C C C C H

=O
OH H OH OH OH
Atom C Chiral

H HO H H H

2 3 4 5 6

6 CH

b
O H
1 1

2OH

H
4

OH H

HO

OH
3

OH

OH

a-D-Glukosa

Atom C asimetris terjauh dari karbonil (CO) Sebagai dasar penamaan D (OH dikanan) dan L (OH di kiri)

a-D-Glukosa
(Aldosa) (Aldohexose)

Struktur Kimia

Struktur Kimia
H
1

C C C C C C H

=O
OH H OH OH OH

H
H HO H H H

1 2 3 4 5 6

C C C C C C H

OH OH H OH O OH
H
4

H HO H H H

2 3 4 5 6

6 CH 5

2OH

O
3 2

H
1

HO

OH H

OH

OH

a-D-Glukosa Konfigurasi Haworth

a-D-Glucopyranosa Aldehydo-D-Glukose

Struktur Kimia
Gugus Aldehid (CHO) = gugus reduksi H
1

Gugus Karbonil (CO)

H
H
1 2 3 4 5 6

C C C C C C H

=O
OH Reduksi H OH OH OH Na-Amalgam Li-Al-hydride hydrogenation

OH

H HO H H H

2 3 4 5 6

H
HO H H

C
C C C C H

OH
H OH OH OH

a-D-Glukosa (Aldosa)

Sorbitol

Struktur Kimia
Gugus Aldehid (CHO) = gugus reduksi H
1

Gugus Karbonil (CO)

H
H
1 2 3 4 5 6

C C C C C C

=O
OH H OH OH OH HO R C= O C H Gugus Keton

OH

H HO H H H

2 3 4 5 6

C= O
C C C C H H OH OH OH

Saccharose group

HO H H

H a-D-Glukosa (Aldosa) (Hexose) (C6H12O6)

R = H Aldehyde R = CH2OH Keton

a-D-Fruktosa (Ketosa) (Hexulose)

(C6H12O6)

Struktur Kimia
Carbonyl group
Aldehydes are readily oxidised to carboxylic acids:

Keton

Aldehid

If copper (II) (blue Cu2+) is used as the oxidising agent it is converted into copper (I) which forms red copper oxide (Cu2O) as a precipitate. This is how Benedicts reagent and Fehlings solution are used to test for aldehydes and reducing sugars.

MONOSAKARIDA
6 CH
2OH

CH2OH O H
1

H
4

HO

O OH H H OH

H OH

HO

OH
3

OH

OH

a-D-Glukosa
6

a-D-Galaktosa
CH2OH

OHCH2 O
5

CH2OH
2

H OH

OH OH 3 H

HO

OH
H

OH H

OH

a-D-Fructose

a-D-Manosa

DISAKARIDA
CH2OH H O H O CH2OH

H
OH

H OH

HO

OH
H

H OH

a-D-Glukosa

OH H a-D-Glukosa

(Glukosa-a-(14)-Glukosa)

a-D-Maltosa

terdiri atas 2 molekul glukosa dari hydrolysis pati oleh b-amylase gula reduksi

DISAKARIDA
CH2OH HO H OH H O H H OH O CH2OH

H
OH

H OH

a-D-Galaktosa

OH H a-D-Glukosa

Galaktose-b-(14)-Glukosa)
terdiri atas Galaktosa dan Glukosa terdapat pada susu : Mamalia : 2 8.5% Cow and Goat : 4.5 4.8% Human : 7 % gula reduksi

Lactosa

DISAKARIDA
CH2OH H O H O
6

OHCH2 O
5

CH2OH
2

HO

OH
H

H OH

OH OH
H

OH

a-D-Glukosa

a-D-Fructose

Glukosa-a-(12)-Fruktosa)
terdiri atas Glukosa dan Fruktosa terdapat pada cane atau beet gula non-reduksi table sugar

Sukrosa

POLISAKARIDA
ikatan-b-(14)-Glikosidik)
H OH
2 1

CH2OH

B1

HO

OH H CH2OH

H O

H
4

O OH H
1

O
H

a-D-Glukosa

OH H a-D-Glukosa

A CELLULOSE

POLISAKARIDA

POLISAKARIDA

Pendahuluan

Evolution of analytical methods for carbohydrates has followed the succession:

qualitative color tests, adaptation of the color test for reducing sugars based on reduction of Cu(II) to Cu(I) (Fehling test) to quantitation of reducing sugars, qualitative paper chromatography, quantitative paper chromatography, gas chromatography (GC) of derivatized sugars, qualitative and quantitative thin layer chromatography, enzymic methods, and highperformance liquid chromatography (HPLC). nuclear magnetic resonance, near-infrared (NIR) spectrometry, antibodies (Immunoassays) Fluorescence spectrometry CE dan MS

Pendahuluan
Pelabelan (FDA): Total carbohydrate calculated by subtraction of the sums of the weights of crude protein, total fat, moisture, and ash in a serving from the total weight of the food in a serving. Other carbohydrate (formerly called complex carbohydrate) is obtained by calculating the difference between the amount of total carbohydrate and the sum of the amounts of dietary fiber and sugars. Sugars are defined as the sum of all free monosaccharides (D-glucose and -fructose) and disaccharides [sucrose, lactose, and maltose (if a maltodextrin or glucose/corn syrup has been added)]

Pendahuluan
Total KH = 100% - (% crude protein + % crude fat + kadar air + kadar abu) KH kompleks = total KH (serat pangan + gula) Serat pangan = jenis KH yang tidak bisa dicerna (selulose, hemiselulose dan polisakaride lain yang berfungsi sebagai bahan pelindung tanaman, serta lignin) Gula = jumlah monosakarida bebas + disakarida/ maltodextrin

Preparasi Sampel
vacuum oven and drying to constant weight at 55C and 1mm Hg pressure

Soxhlet extractor

Removed contaminants

Mono- dan Oligosakarid a

Metode analisis

Total karbohidrat Total reducing sugar

Phenol-sulfuric method SomogyiNelson method HPLC

GC

Enzimatis Metode khusus

MS

TLC

CE

Total Carbohydrate: PhenolSulfuric Acid Method

Prinsip KH didigesti dgn asam dan atau suhu panas, tjd reaksi dehidrasi, menghasilkan turunan furan. Lalu dikondensasi dgn senyawa fenolik (fenol, resorsinol, naphtol) dan amin aromatis (anilin)

Total Carbohydrate: PhenolSulfuric Acid Method Prosedur umum :

1. 2. 3.

4.

5.

A clear, aqueous solution of carbohydrate(s) is transferred using a pipette into a small tube. A blank of water also is prepared. An aqueous solution of phenol is added, and the contents are mixed. Concentrated sulfuric acid is added rapidly to the tube so that the stream produces good mixing. The tube is agitated. (Adding the sulfuric acid to the water produces considerable heat.) A yellow-orange color results. Absorbance is measured at 490 nm. The average absorbance of the blanks is subtracted, and the amount of sugar is determined by reference to a standard curve. D-glucose is used to prepare the standard curve.

Total Reducing Sugar: SomogyiNelson Method

Prinsip terjadi reaksi reduksi-oksidasi. Gula pereduksi yaitu gula yg mengandung gugus aldehid (donor elektron/ reduktor) mengalami oksidasi, menghasilkan gugus asam karboksilat. Gugus keton merupakan gula pereduksi yg lemah. The SomogyiNelson method is based on the reduction of Cu(II) ions to Cu(I) ions by reducing sugars.

Total Reducing Sugar: SomogyiNelson Method

Prosedur umum: 1. A solution of copper(II)

2. 3.

4.

5.

sulfate and an alkaline buffer are added by pipettes to a solution of reducing sugars(s) and a water blank. The resulting solution is heated in a boiling water bath. A reagent prepared by mixing solutions of acidic ammonium molybdate and sodium arsenate is added. After mixing, dilution, and remixing, absorbance is measured at 520 nm. After subtracting the absorbance of the reagent blank, the A250 is converted into glucose equivalents using a standard plot of micrograms of glucose vs. absorbance.