IDENTIFYING

MACROMOLECULES IN
Nutrient
LAB

Inthislab,withtheuseof
indicatorsaschemical
detectiontools,youwill
analyzeavarietyoffoods
forthepresenceof
nutrients.
Detectionisbasedupon
observingachemical
changethattakesplace
mostoftenachangein
color.

LabObjective
Identifythepresenceofmajornutrientssuchas
simplecarbohydrates(glucose),complex
carbohydrates(starch),proteinandfatin
commonfoods.

Whatisanindicator?
Indicatorsarechemicalcompoundsusedto
detectthepresenceofothercompounds.

BackgroundInformation
INDICATOR

MACROMOLECULE

NEGATIVE POSITIVE
TEST
TEST

Benedicts
solution

simple
carbohydrate

blue

orange

Iodine
solution

complex
carbohydrate

dark red

black

Biuret
solution

protein

blue

violet,
black

Sudan IV

lipid

dark red

reddishorange

TestforSimpleCarbohydrates
Benedictssolution
Benedict'ssolutionisachemicalindicatorforsimple
sugarssuchasglucose:C6H12O6.
Aquablue:negativetest;yellow/green/brick
red,etc.:positivetest

TestforSimpleCarbohydrates
Benedictssolution
Unlikesomeotherindicators,Benedicts
solutiondoesnotworkatroomtemperature
itmustbeheatedfirst.

TestforComplexCarbohydrates
IodineSolution
Iodinesolution colorchange=bluetoblack

TestforComplexCarbohydrates
Iodinesolution
Iodinesolutionisanindicatorforamoleculecalled
starch.
Starchisahugemoleculemadeupofhundredsof
simplesugarmolecules(suchasglucose)connected
toeachother.

TestforProtein(aminoacids)
Biuretsolution
Biuretsolution darkvioletbluetopinkish
purple

TestforFats(lipids)
SudanIV
IflipidsarepresenttheSudanIVwillstainthem
reddishorange(positivetest).

Question
Whydidntthetesttubecontainingsucrose
changecolors?

Question
Whydidntthetesttubecontainingstarch
changecolors?

ReducingandNonReducingSugars
Reductionisthechemiststermforelectrongain
Amoleculethatgainsanelectronisthus
reduced

Amoleculethatdonateselectronsiscalleda
reducingagent

Asugarthatdonateselectronsiscalleda
reducingsugar

Theelectronisdonatedbythecarbonylgroup
Benedictsreagent changescolourwhenexposedto
areducingagent

BenedictsTest
Benedictsreagentundergoesa
complexcolourchangewhenit
isreduced
Theintensityofthecolour
changeisproportionaltothe
concentrationofreducingsugar
present
Thecolourchangesequenceis:

Blue
green
yellow
orange
brickred

Thecarbonylgroup monosaccharides
Thecarbonylgroupisfree
inthestraightchainform
Butnotfreeintheringform
BUTremember thering
formandthestraightchain
formareinterchangeable
Soall monosaccharidesare
reducingsugars
Allmonosaccharidesreduce
Benedictsreagent

Thecarbonylgroup
disaccharides maltose
Insomedisaccharides
e.g.maltoseoneofthe
carbonylgroupsisstill
free
Suchdisaccharidesare
reducingsugars
TheyreduceBenedicts
reagent

Thecarbonylgroup
disaccharides sucrose
Insomedisaccharidese.g.
sucroseboth ofthe
carbonylgroupsare
involvedintheglycosidic
bond
Sotherearenofree
carbonylgroups
Suchsugarsarecallednon
reducingsugars
TheydoNOT reduce
Benedictsreagent

Thecarbonylgroup
disaccharides sucrose
Thesubunitsofsucrose
(glucoseandfructose)are
reducingsugars
Ifsucroseishydrolysedthe
subunitcanthenactas
reducingsugars
Thisisdoneinthelabby
acidhydrolysis
Afteracidhydrolysissucrose
willreduceBenedicts
reagent