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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
O
HO
HO
OH
OH
OH
HO
OH
O
HO OH
OH
Fructose Glucose
O
HO
OH
OH
OH
Galactose
O
HO
HO OH
OH
Mannose
OH
OH
etc
O
HO
HO
HO
OH
Sucrose
HO
OH O
HO
OH
O
O
HO
OH
OH
OH
O
HO
OH
OH
OH
O
O
HO
OH
OH
OH
O
HO
HO
OH
OH
O
O
HO
OH
OH
OH
O
HO
OH
OH
O
HO
Lactose Maltose
Cellobiose
etc
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Common mono- and di-saccharides
O
HO
HO
OH
OH
OH
HO
OH
O
HO OH
OH
Fructose Glucose
O
HO
OH
OH
OH
Galactose
OH
O
HO
HO
HO
OH
Sucrose
HO
OH
O
HO
OH
O
O
HO
OH
OH
OH
O
HO
OH
OH
OH
O
O
HO
OH
OH
OH
O
HO
HO
OH
OH
O
Lactose Maltose
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Common mono- and di-saccharides
O
HO
HO
OH
OH
OH
HO
OH
O
HO OH
OH
Fructose Glucose
O
HO
OH
OH
OH
Galactose
OH
O
HO
HO
HO
OH
Sucrose
HO
OH
O
HO
OH
O
O
HO
OH
OH
OH
O
HO
OH
OH
OH
O
O
HO
OH
OH
OH
O
HO
HO
OH
OH
O
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
O
HO
HO
OH
OH
OH
Trimethylsilyl
chloride
Methylsilyl
ether
Primary amine
Glycosylamine
Ac
2
O
Acetate
(LCAnalysis)
(GCAnalysis) (LCAnalysis)
(GCAnalysis)
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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
Lactose Purity of lactose 984.22
Glucose, fructose Raw cane sugar 2000.17
Glucose, fructose, sucrose,
lactose
Cane & beet final
molassess
996.04
Glucose, fructose, sucrose,
maltose
Presweetened cereals 982.14
Fructose, glucose, lactose,
maltose, sucrose
Milk chocolate 980.13
Fructose, glucose, sucrose Honey 977.20
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Current GB LC Method
GB-T 22221-2008


Determination of fructose, glucose, sucrose,
maltose, lactose in foods high performance
liquid chromatography
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Mono- and di-saccharidesanalysis
Amine column
Acetonitrile/water (80:20)
Refractive index detector
2 mL/min at 30
o
C
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Mono- and di-saccharidesanalysis
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-saccharidesanalysis
Shodex column
Water
Refractive index detector
0.5 mL/min at 80
o
C
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Mono- and di-saccharidesanalysis
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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
HO
HO
OH
OH
OH
OH
HO
HO
OH
O
OH
R H
O
CuO
R OH
O
+ Cu
2
O
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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
low molecular weight
carbohydrate of animal diet).
NOT appropriate for
testing general food!!
O
HO
HO
HO
OH
HO
OH
O
HO
OH
O
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Typical analysis
Sugars in
sample
Sugars in
sample
Preparation/
Clean up
Preparation/
Clean up
LC
LC
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Mono- and di-saccharidesanalysis
Extract the
sugars from
sample using a
mixture of ethanol
and water
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Mono- and di-saccharidesanalysis
Centrifuge the
mixture after the
extraction
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Mono- and di-saccharidesanalysis
Clean up the
extract by SPE
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Mono- and di-saccharidesanalysis
Clean up the
extract by SPE
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Mono- and di-saccharidesanalysis
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