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Analysis of Sugars

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Definition

„ Nutritional labelling of food products requires


listing of sugars content.

„ Sugars means all mono-saccharides and


di-saccharides present in food*.

*Codex Guidelines on Nutrition Labelling, CAC/GL 2-1985

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Mono- and di-saccharides
HO OH OH OH OH
OH
O OH OH
HO
O O O etc
HO
OH HO OH HO OH HO OH
HO OH OH

Fructose Glucose Galactose Mannose

OH

O OH
HO OH OH
HO OH O
HO HO
O OH
O HO
HO O OH
HO O O
O OH HO OH
OH OH O
HO OH
OH

HO OH

Sucrose Lactose Maltose


OH OH

O O
HO
HO O
HO OH
etc
OH OH

Cellobiose

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Common mono- and di-saccharides
HO OH OH OH OH
O OH
O O
HO
OH HO OH HO OH
HO OH OH

Fructose Glucose Galactose

OH

O OH
HO OH OH
HO OH O
HO HO
O HO OH
O
HO O OH
HO O O
O OH HO OH
OH OH O
HO OH
OH
HO OH

Sucrose Lactose Maltose

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Common mono- and di-saccharides
HO OH OH OH OH
O OH
O O
HO
OH HO OH HO OH
HO OH OH

Fructose Glucose Galactose

OH

O OH
HO OH OH
HO OH O
HO HO
O HO OH
O
HO O OH
HO O O
O OH HO OH
OH OH O
HO OH
OH
HO OH

Sucrose Lactose Maltose

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Common sugars in foods

„ Mono-saccharides:

Fructose, glucose, galactose

„ Di-saccharides:

Lactose, maltose, sucrose

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Properties of sugars
(LC Analysis)

OH Trimethylsilyl
Primary amine chloride
Glycosylamine O Methylsilyl
HO ether
(LC Analysis) HO OH (GC Analysis)
OH

Ac2O

Acetate
(GC Analysis)

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Properties of sugars

„ Water soluble.

„ Low thermal stability.

„ Do not have properties of fluorescence


emission.

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Sugar Analysis

LC Methods with:

„ Refractive Index Detector


„ Evaporative Light Scattering Detector
„ Pulsed Amperometric Detector

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Current AOAC LC Methods
977.20 Honey Fructose, glucose, sucrose

980.13 Milk chocolate Fructose, glucose, lactose,


maltose, sucrose
982.14 Presweetened cereals Glucose, fructose, sucrose,
maltose

984.22 Purity of lactose Lactose

996.04 Cane & beet final Glucose, fructose, sucrose,


molassess lactose
2000.17 Raw cane sugar Glucose, fructose

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Current GB LC Method

GB-T 22221-2008
„ 食品中果糖、葡萄糖、蔗糖、麦芽糖、乳糖的测
定 高效液相色谱法

„ Determination of fructose, glucose, sucrose,


maltose, lactose in foods – high performance
liquid chromatography

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Mono- and di-saccharides analysis
Amine column
Acetonitrile/water (80:20)
Refractive index detector
2 mL/min at 30 oC

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Mono- and di-saccharides analysis

„ Amine column may not be able to resolve all


the peaks.

„ Alternative column may be used, e.g.,


Shodex sugar column.

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Mono- and di-saccharides analysis

Shodex column
Water
Refractive index detector
0.5 mL/min at 80 oC

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Mono- and di-saccharides analysis

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Analysis of reducing sugars

AOAC methods:

920.183 Sugars (Reducing) in Honey


920.190 Sugars (Reducing) in Maple
Products
945.66 Total Reducing Sugars
etc.

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Analysis of reducing sugars

„ Fehling solution is used in the methods

„ It relies on chemical reaction between the


sugars and the Fehling solution.
O O
CuO
+ Cu2O

R H R OH

OH OH

O OH
HO HO
HO OH HO O
OH OH

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Limitation of analyzing reducing sugar

„ The best-known and most significant


non-reducing sugar in foods is sucrose!
(The most important OH

low molecular weight HO


O

HO
carbohydrate of animal diet). HO

HO O
O OH

NOT appropriate for


HO OH
testing general food!!

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Typical analysis

Sugars
Sugars in
in Preparation/
Preparation/ LC
LC
sample
sample Clean
Clean up
up

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Mono- and di-saccharides analysis

Extract the
sugars from
sample using a
mixture of ethanol
and water

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Mono- and di-saccharides analysis

Centrifuge the
mixture after the
extraction

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Mono- and di-saccharides analysis

Clean up the
extract by SPE

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Mono- and di-saccharides analysis

Clean up the
extract by SPE

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Mono- and di-saccharides analysis

Analyse the
extract using LC
with refractive
index detector

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Proficiency test

FAPAS

LGC

AOAC

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CRM

BCR

NIST

LGC

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Points to note

Sugar alcohols (polyols) are not “sugars”

Not all monosaccharides and disaccharides


are reducing sugars. Test method for reducing
sugar may not be suitable for the analysis
of sugars.

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Points to note

Definition of “0”
Sugars ≤0.5 g/100 g

Limit of detection of total sugars should


be less than or equal to 0.5 g/100 g.

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Thank You

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