ErikaHaase

EyramandEnyoFiadonu,KateLeJeune
MsSloan
Period2
8/24/15
TestingforPresenceofMacromolecules

*SeelabhandoutforPart1Procedure*

DataPart1:

Benedict

Iodine

Biuret

Ninhydrin

SudanIII

Control
(Water)

Clear,
Negative

Yellow,
negative

Reddish,
negative

Reddish,
negative

Clear,
negative

Solution1

Clear,
negative

Darkblue,
positive

Clear,
negative

Clear,
negative

Littlered,
positive

Solution2

Orange,
positive

Yellow,
negative

Clear,
negative

Purple,
positive

Clear,
negative

Solution3

Purple,
negative

Yellow,
negative

Purple,
positive

Clear,
negative

Reddish,
negative

Solution4

Separated,
negative

Yellow,
negative

Clear,
negative

Reddish,
negative

Redand
separated,
positive

DataSummaryPart1
Thecontrolgroupwasnegativeforalltheindicators.Solution1mostlikelycontained
polysaccharides.Solution1testedpositivewiththeiodineindicator,turningthesolutiondark
blue,andthesudanIIIindicator,turningitslightlyred.Solution2mostlikelycontained
monosaccharidesittestedpositivewiththebenedicttest,turningorange.Solution3mostlikely
containedproteinsandaminoacidsbecauseittestedpositiveforboththebiuretandninhydrin
indicators,turningthesolutionpurple.Finally,solution4testedpositivewiththesudan
indicator,turningred,suggestingitcontainslipids.

ProcedurePart2:
1. Label10testtubeswithmaskingtape(controlofundigestedfood#15anddigestedfood
#15)andplaceintesttuberack.

BenedictTest
1. Add1mLofyogurttothebottomofeachtesttubeanddilutewith2mLofwater.
2. Add25spitsofamylasetooneofthetesttubesandlabelthistubedigested.(Note:Let
sitsotheamylasecanbreakdownthefood)
3. Add5dropsofBenedictsreagentintoeachofthetesttube.
4. Gentlymixeachtesttube.
5. Heatthetesttubeinahotwaterbathforatleast2minutes.
6. Ifamonosaccharideispresentthesolutionwillturnyellowororange.Abluesolution
willbenegative.
7. Recorddataindatatable.
8. Washtesttubethoroughlywithsoapandwater.
IodineTest
1. Add1gofbreadinthetesttubeanddilutewith1mLofwater.
2. Add5dropsofiodinetesttubeandgentlyswirl.
3. Add25spitsofamylasetooneofthetesttubesandlabelthetubedigested.(Note:Let
sitsotheamylasecanbreakdownthefood)
4. Gentlymixeachtesttube.
5. Ifapolysaccharideispresentthesolutionwillturnblackordarkblue,signalinga
positivereaction.Ifthetestisnegative,thesolutionwillremainbrown/yellow.
6. Recorddataindatatable.
7. Washtesttubethoroughlywithsoapandwater.

BiuretTest
1. Add1mLofyogurttothebottomofeachtesttubeanddilutewith2mLofwater.
2. Add0.5mLofpepsintooneofthetesttubesandlabelthetubedigeste
3. Add5dropsofBiuretsolutiontothetubeandmixgently.
4. Thebiuretsolutionwillcauseacolorchangetopurpleifthereactionispositive.Ifthe
reactionisnegative,thesamplewillremainblue.
5. Recorddataindatatable.
6. Washtesttubethoroughlywithsoapandwater.

NinhydrinTest
1. Add1mLofyogurttothebottomofeachtesttubeanddilutewith2mLofwater.
2. Add0.5mLofpepsintooneofthetesttubesandlabelthetubedigested.
3. Add10dropsofNinhydrintothetubeandmixgently.
4. Gentlymixthesolutionsandputinboilingwaterfor5minutes.Letsitforabitand
recordcolor.SincethistestistestingforaminoacidsandproteinswithfreeNH2.When
suchagroupreactswithninhydrin,apurplebluecolorisformed.
5. Inyourdatatable,noteifthetestwaspositiveornegative,andthefinalcolor.

6. Washtesttubethoroughlywithsoapandwater.

SudanIII
1. Add1mLofyogurttothebottomofeachtesttubeanddilutewith1mLofwater.
2. Add0.5mLofbiletooneofthetesttubesandlabelthetubedigested.
3. Add25dropsofSudanIIItoeachtesttubeandmixgently.
4. Iflipidsarepresent,SudanIIIwilldissolvethelipidsandstainthemred.
5. Recorddataindatatable.
6. Washthetesttubethoroughlywithsoapandwater.

Prediction:

Control(undigestedfood)

Yogurtdigested

Benedict

Orange/Positive

Orange/Positive

Iodine

DarkBlue/Positive

DarkBlue/Positive

Ninhydrin

nocolorchange/negative

Purple/positive

Biuret

Purple/positive

Purple/positive

SudanIII

Red/positive

nocolorchange/negative

DataPart2:

Bread
Benedict

Yogurt

Control(Undigested
Food)

Nottested

Orange/white,positive

Orange/white,positive

Brown/yellow,neg

Brown/yellow,negative

Yogurt
Brown/yellow,negative
Bread
Black/darkblue
positive

Ninhydrin

Nottested

Darkpurple,positive

Lightpurple,positive

Biuret

Nottested

Darkerpurple,positive

Lighterpurple,positive

Iodine

SudanIII

Nottested

Notasmanyredglobsbutstill Moreredfatglobules,
visible,positive
positive

DataSummaryPart2
Thecontrolortheundigestedgroupfortheyogurttestedpositivewiththebenedict,ninhydrin,
biuret,sudanIII,andtestednegativeforiodine.Thedigestedyogurttestedpositiveforbenedict,
ninhydrin,biuret,andlessforsudanIII.Theundigestedbreadtestednegativeforiodine,The
digestedbreadtestedpositiveintheiodinesolution.