ITP301

FOOD ANALYSIS LABORATORY

2(0-2)

Food Technology Program

Departement of Food Science and Technology
Faculty of Agricultural Technology
IPB University

http://fst.ipb.ac.id
 Internationally Recognized Undergraduate Program by IFT & IUFoST

Topic 6

Sample preparation for sugar analysis

Didah Nur Faridah

Food Technology Program

Departement of Food Science and Technology, FATETA-IPB
Aim of the laboratory practice
§ This practice is aimed to explain and practice
the principles of sample preparation for sugar
analysis.
Learning Outcomes
After completing this laboratory work, students are
expected to be able to:
§ Explain the principles of sample preparation for
sugar analysis.
§ Perform the analysis of sample preparation for sugar
analysis.
Sample preparation for sugar analysis
§ Analysis of sugar in food requires preparation step to
separate the sugars from food matrices. The results
from sample preparation can be used for analysis of
total sugar content.
§ Sample need to be separated from other
components, such as nitrogen, lipid, phenolic
compounds, and soluble pigments. Those
compounds can interfere filtration or join react and
intrusive measurement of sugar. Therefore,
separation of sugar from the interrupting
compounds before analysis is needed to be done.
Sample preparation for sugar analysis
§ Alcohol or dissolved gases (in carbonated or
fermented products) are generally removed by
evaporation at low temperature. Evaporation in a
vacuum is desirable to prevent the unraveling of
sugar upon heating.
§ Chlorophyll, carotenoids and lipids separated by
extracted with petroleum ether in which sugar is not
dissolved. Pigments can also removed using
activated carbon or lead acetate. The excess of Pb-
acetate can be removed by the addition of Na/K-
oxalate.
Sample preparation for sugar analysis
§ Protein, that may interfere sugar determination with
reduction and colorimetric method, can be
eliminated by precipitation. By adding ethanol or
acetone, protein will agglomerate and can be
separated by filtration or sentrifugation. Protein can
also be precipitated by heavy metals, such as
Zn(OH)2.
Sample preparation for sugar analysis
§ Liquid sample: sample must be made to be alkaline
by addition of CaCO3 so that the acids contained in
the sample did not hydrolyze existing sugar during
heating. Heating is required to inactivate sugar
hydrolyzing enzymes.
§ Solid sample: sample is extracted using alcohol 80%
to separate sugars from other components. Most
sugars are sensitive to alcohols with high
concentrations, so alcohol should be removed with
low heating.
Experimental Procedure

Equipments
§ Analytical balance
§ Beaker glass 300-mL
§ Volumetric flask 100-mL, 250-mL
§ Hot plate
§ Pippette mohr 5-mL
§ Filter paper
§ Funnel
§ Volumetric pipette 5-mL, 10-mL
Experimental Procedure

Reagents
§ CaCO3
§ Saturated Pb-acetate
§ Na-oxalate
§ Alcohol 80%
§ Aquadest
Experimental Procedure

Samples
§ Thick syrup
§ Sweetened condensed milk
§ Honey
§ Strawberry jam
§ Sweet soy-sauce
§ High Fructose Syrup
Experimental Procedure
Procedures
Preparation for Liquid Sample
§ Put 0.1 g sample into beaker glass 300-mL, add 100
mL aquadest and 1 g CaCO3.
§ Boil sample for 30 minutes then chilled.
§ Move the sample into volumetric flask 250-mL.
§ Add 1.5–2.5 mL saturated Pb-acetate solution (until
the solution becomes clear).
§ Add aquadest until volume.
§ Shake and strain it through a filter paper.
Experimental Procedure
§ Take 30 mL of filtrat into other beaker glass. Add 1.5
g dried Na-oxsalate to precipitate Pb.
§ Strain sample. Dilute the filter 20 times. Take 10 mL
filtrate for analysis of total sugar and take 40 mL for
analysis of reducing sugar.
Experimental Procedure
Preparation for Solid Sample
§ Put 20 g sample into beaker glass and add 20 mL of
alcohol 80%.
§ Destruct sample using waring blender.
§ Strain sample through cotton/filter paper and move
filtrate to another beaker.
§ Wash solid residue in cotton with 80% alcohol until
all the sugar dissolved into the filtrate phase.
Experimental Procedure
§ Measure pH value of sample. If acid, add CaCO3 until
alkaline and heated 100° C at water bath for 30
minutes.
§ Chill sample and strain through filter paper
Whatman No. 2.
§ Evaporate alcohol from sample by heating the
filtrate at 85°C in water bath or with a vacuum.
§ Filter back if precipitation still occurs.
§ Add 1.5–2.5 mL saturated Pb-acetate solution (until
the solution becomes clear).
Experimental Procedure
§ Move the filtrate to 250 mL volumetric flask. Add aquadest
until volume.
§ Shake sample, strain, and take 50 mL of filtrate.
§ Add 2.5 g dried Na-oxsalate to precipitate Pb and strain
sample.
§ Take 10 mL filtrate for analysis of total sugar and take 40 mL
filtrate for analysis of reducing sugar. Store in refrigerator.
§ Move 5 mL filtrate for analysis of total sugar into 100 mL
volumetric flask. Add aquadest until volume and shake it.
§ Store sample in refrigerator if not analyzed directly.
§ Do appropriate dilution if needed (if the sugar concentration
is too high). Record the dilution factor used.
References
§ Apriyantono, A., D. Fardiaz, N.L. Puspitasari, S. Yasni and S. Budiyanto.
1989. Petunjuk Praktikum Analisis Pangan. PT IPB Press, Bogor.
§ Fardiaz, D., N. Andarwulan, C.H. Wijaya and N.L. Puspitasari. 1992. Teknik
Analisis Sifat Kimia dan Fungsional Komponen Pangan. Pusat Antar
Universitas Pangan dan Gizi, IPB, Bogor.
§ James, C.S. 1999. Analytical Chemistry of Foods. Aspen Publishers,
Maryland.
§ Nielsen, S.S. 2003. Food Analysis. 3rd ed. Kluwer Academic/Plenum
Publishers, New York.
§ Nielsen, S.S. 2003. Food Analysis Laboratory Manual. Kluwer
Academic/Plenum Pub-lishers, New York.
Contact:
Feri Kusnandar
Email: fkunsnandar@apps.ipb.ac.id