Biochemical Composition of Normal Urine

Biochemicalcompositionof
Nature Precedings : doi:10.1038/npre.2011.6595.1 : Posted 12 Nov 2011
normalurine
AtifAminBaig
Lecturer,
FacultyofMedicineandHealthSciences,
UniversitiSultanZainalAbidin
TotalBodyWater(TBW)=60%ofbodyweight
Highestinnewbornsandadultmales
lowestinadultfemalesandinadultswithalargeamountofadiposetissue
TotalBodyWater
2/3 1/3
K+ Na+
Mg+2 ClandHCO3
Intracellular
Proteinorganicphosphates Extraracellular
(ATP,ADPorAMP)
1/4 3/4
Albumin
globulin Plasma Interstitialfluid
Functionofkidney
Controllingthebloodvolumeandcomposition
HumanMetabolicWastes
Thebyproductofallchemicalreactionsinthebodythat
needs to be excrete or else the build up of these
compoundswillsickenandeventuallykilltheperson.
Metabolicwastes
CO2 Nitrogenouswastes
Respiratorysystem
Urinarysystem Urinarysystem
Nitrogenouswastes
Proteinmetabolism 2NH3+CO2H2NC0NH2 Urea
Nucleicacidmetabolism Majorportionfrompurines Uricacid
Creatine
azotemia Creatinine
Uremia
Ureacomprised50%oftotalnitrogenouswastesinthehumans
Urine
Definition
Bydefinitionurineisa?
Aliquidcontainingmultiplewasteproductsof
metabolism,especiallyureaandothernitrogenous
compounds,thatarefilteredfromthebloodbythe
kidneys.Urineisstoredintheurinarybladderandis
excretedfromthebodythroughtheurethra.
Ultrafiltrationofplasma
Ultrafiltrationoccursthroughglomerulusgivingrisetoprimaryurine
Theprimaryurineisisotonicwithplasma.
Anionicmoleculesarerepelled
CollagenTypeIV
bythenegativechargeofthebasementmembrane
Poresizewith
meandiameterof
2.9nm
Plasmacomponent
morethan64KDaare Plasmacomponent
totallyblocked lesserthan15KDa
C.Inorganicconstituents
Urinalysis
Theroutineurineexamination.
Mostusefultoolforthecliniciansasanindicatororhealthor
disease.
Particularly,usedinrenalmetabolicdisorders.
Oftendoneforpatientsadmittedtothehospital.
Theroutineurineanalysisisdividedintofourmaingroups:
1. PhysicoChemicalproperties.
2. Chemicalexamination.
3. Microscopicexamination.
4. Bacterialscreening
1.PhysicoChemicalproperties 2.Chemicalexamination
Color pH
Appearance Sugar
Volume Protein
Specificgravity Ketonebodies
Odor Bilirubin
Urobilinogen
Occultblood
Nitrite
Ascorbicacid
3.Microscopicexamination
WBCs
Epithelialcells
RBCs 4.Bacterialscreening
Casts Gramstaining
Crystals Colonycount
Bacteria
Yeasts
Nonbacterialorganisms
Urinecollection
Urinemustbecollectedinasterilebottle.
Theroutineurineexaminationmustbe
conductedwithin30min.
Incaseofdelay,urinemustberefrigerated.
Delayinsampleexaminationwillresultin:
DecreasedpHbytheutilizationofglucosebybacteria.
IncreasedpHbytheconversionofureatoammoniabybacteria.
InincreasedpH,thetendencyofphosphatestoprecipitatewill
increase.
Oxidationofurobilinogentourobilin.Itmaygivefalsenegative
resultforjaundice.
Typesofspecimen
Random First Afternoon 24hours

morning
Chemical& Usedforclearance
microscopic testofkidney
Specimenof Takenbetween24
examination choice pm
Samplesare
Takenanytime collectedfor24
Most Bestfordetecting
afterfirstmorning hours(foreg,
concentrated urobilinogen
urine 8amtonextday8
am).
Microscopicand
Night Day
chemical Postprandial
Itistoavoidany
examination
changesinthe
Allurinesamples Allurinesamples Takenafter2hoursof dilutionofvarious
arecollectedin12 arecollectedin meal suspended
hourofnight(for 12hourofday constituentsof
eg,8pmto8am) (foreg,8amto urine
8pm)
PhysicoChemicalproperties
Colorofnormalurine
Normalurinecolorrangesfrompaleyellowtodeep
amber.
itistheresultofapigmentcalledurochrome.
BvitaminsturnurineaneyepoppingneonyellowBUTmayalso
indicateliverdisease.
porphyria,adiseasethataffectsyourskinandnervoussystem,
turnsurinethecolorofportwine.
Colorofnormalurine
Mostchangesinurinecolorareharmlessandtemporary
andmaybedueto:
Certainfoodsbeetsmayturnurinered
Dyesinfoods/drinks
Supplementsvitamins
Prescriptiondrugs
Unusualurinecolorcanindicateaninfectionorserious
illness.
Different colors of urine

pale yellow (straw)
light yellow
yellow
green-yellow (olive)
red-yellow
red
red-brown
brown-black
black
milky
Clarityofurine
Appearanceofnormalurine
Appearanceofurine
Substancesthatcausecloudinessbutthatare
notconsideredunhealthyinclude:
mucous,
spermandprostaticfluid,
cellsfromtheskin,
normalurinecrystals,and
contaminants(likebodylotionsandpowders).
Othersubstancesthatcanmakeurinecloudy
(suchasredbloodcells,whitebloodcells,or
bacteria)indicateaconditionthatrequires
attention.
Volumeofurine
Normalvolumeis7502000ml/24H
Under 750 ml (oligouria) dehydration, infection, obstruction, renal

stones,kidney,failure,etc).
Over 2000ml (polyuria) diabetes insipidus, hypertension,

nephroticsyndrome, ingestion of alcohol or drugs, endocrinal
disorders.
Absentofurine(anuria)obstruction,kidneyfailure,stenosis.
Odorofurine
Healthyurinemayhaveamildsmellbutgenerallydoes
nothaveafoulodor.
Insomecases,anunusualorstrongurineodormaybe
duetobenignconditionsthatarenotharmful,suchas
eatingcertainfoodsortakingcertainmedicationsand
volatileacids.
Insomecases,anunusualorstrongurineodormaybe
duetobenignconditionsthatarenotharmful,suchas
eatingcertainfoodsortakingcertainmedications.
Specificgravity
Specific gravity reflects kidney's ability to concentrate.

Want concentrated urine for accurate testing, best is first morning
sample.
Low specimen not concentrated, kidney disease.
High first morning, certain drugs
Random specimen (1.003 to 1.032)
24 hours specimen (1.015 to 1.025)
Severe renal tubular damage, sp. Gravity is fixed at 1.010 isothenuric
Osmolalityofurine
Determination of urinary osmolality is more important

than specific gravity because it gives more accurate
reflectionoftheconcentrationofdissolvedsubstances.
Itisthemeasureofthemolesofdissolvedparticles(un
dissociatedmoleculesandions)containedinakilogram
(Kg)ofasolvent.
The normal osmolality of random urine specimen varies from 401350

m,osmol/kg.
The normal osmolality of 24 hours urine specimen varies from 500800
m,osmol/kg.
ChemicalExamination
Reagent strips are used only once and discarded.

Testing
Perform within 1 hour after collection
Allow refrigerated specimens to return to room
temperature.
Dip strip in fresh urine and compare color of pads
to the color chart after appropriate time period.
Instruments are available which detect color
changes electronically
UsingReagentStrips
BRIEFLY dip the strip in urine.

Colors are matched to those on the bottle label at the
appropriate times.
Timing is critical for accurate results.
ReagentStrips
Glucose
Presence of glucose (glycosuria) indicates that the blood

glucose level has exceeded the renal threshold.
Useful to screen for diabetes.
Bilirubin
Bilirubin is a byproduct of the breakdown of hemoglobin.

Normally contains no bilirubin.
Presence may be an indication of liver disease, bile duct obstruction or
hepatitis.
Since the bilirubin in samples is sensitive to light, exposure of the
urine samples to light for a long period of time may result in a false
negative test result.
Ketones
Ketones are excreted when the body metabolizes fats

incompletely (ketonuria)
Blood
Presence of blood may indicate infection, trauma to the

urinary tract or bleeding in the kidneys.
False positive readings most often due to contamination
with menstrual blood.
Ph
pH measures degree of acidity or alkalinity of urine

Protein
Presence of protein (proteinuria) is an important indicator

of renal disease.
False negatives can occur in alkaline or dilute urine or
when primary protein is not albumin.
Urobilinogen
Urobilinogen is a degradation product of bilirubin formed

by intestinal bacteria.
It may be increased in hepatic disease or hemolytic disease
Nitrite
Nitrite formed by gram negative bacteria converting

urinary nitrate to nitrite
Leukocytes
Leukocytes (white blood cells) usually indicate infection.

Leucocyte esterase activity is due to presence of WBCs in
urine while nitrites strongly suggest bacteriuria.
NormalValues
Negative results for glucose, ketones, bilirubin, nitrites,

leukocyte esterase and blood.
Protein negative or trace.
pH 5.5-8.0
Urobilinogen 0.2-1.0 Ehrlich units
HandlingandStorageof
Strips
Handling and Storage

Keep strips in original container
Do not touch reagent pad areas
Reagents and strips must be stored properly to
retain activity
Protect from moisture and volatile fumes
Stored at room temperature
Use before expiration date
Procedure
Dipstripbriefly,butcompletelyintowellmixed,room
temperatureurinesample.
Withdrawstrip.
Blotbrieflyonitsside.
Keepthestripflat,readresultsattheappropriatetimesby
comparingthecolortotheappropriatecoloronthechart
provided.
SourcesofError
TimingFailuretoobservecolorchangesatappropriate
timeintervalsmaycauseinaccurateresults.
Lighting Observe color changes and color charts

undergoodlighting.
QC Reagent strips should be tested with positive

controlsoneachdayofusetoensureproperreactivity.
Sample Proper collection and storage of urine is

necessarytoinsurepreservationofchemical.
SourcesofError
Testing cold specimens would result in a slowing

down of reactions; test specimens when fresh or bring
themtoRTbeforetesting
Inadequate mixing of specimen could result in false

reduced or negative reactions to blood and leukocyte
tests;mixspecimenswellbeforedipping
Overdippingofreagentstripwillresultinleachingof
reagents out of pads; briefly, but completely dip the
reagentstripintotheurine
Parameter Normal Abnormal
Colour Yellowstraw, Darkamberconcentratedurineinsufficientoffluidintake
amber, Cloudyinfectiousprocess
transparent Darkorangedrug,eg:pyridium
Redordarkbrowndiseaseprocesscausingbloodin
urine
Consistency Clearliquid Mucousplug,viscid,thickinfectiousprocess
Odor Faintaromatic Offensiveinfectiousprocess
Sterility No MicroorganismpresentinfectiousofUT
microorganism
present
pH 4.58.0 Overthan8.0UTinfection
Under4.5uncontrolleddiabetes,starvation,dehydration
Specificgravity 1.010to1.025 Over1.025diabetesmellitus,underhydration
Under1.01diabetesinsipidus,kidneydisease,over
hydration
Urineglucose Notpresent Presentdiabetesmellitus
Urineketone Notpresent Presentdiabeticcoma,starvation,prolongedvomiting
bodies(eg:
acetone)
Urineblood Notpresent Occultkidneydisease
Brightredhemorrhage
Urinemicroscopy
Whitebloodcells
Pyuriareferstothepresenceofelevatednumberof
leukocytes(granulocytes):
UpperorlowerUTinfection
Glomerulonephrotis
Vaginal&cervicalinfections
Externalurethralmeatus(men&women)
Normalrange:
02/HPF(5+indicatesaninfectionand10+indicatesmoresevere
conditions).
Urinemicroscopy
Epithelialcells
Normalurinenotpresent/unquantified
Renal tubular epithelial cells, usually larger than

granulocytes, contain a large round or oval nucleus
arenormallysloughintotheurineinsmallamounts.
Positive nephrotic syndrome, condition leading to

tubulardegeneration.
Urinemicroscopy
Urinarycrystals
Normalurinepresentbutunquantified.
Typeofcrystals:
Calciumoxalate
Triplephosphate
Amorphousphosphates
Uricacids
Uricacidcrystals(hyoeruricemia)
Cyctine
Tyrosine
Leusine
Elevatednumbersofcrystalmayindicates:
Hipercalcemia
Cystinecrystalcystinuria
Mayduetorenalfailureorotherrenaldisease.
Urinemicroscopy
Casts
Formedatthedistalconvolutedtubuleorthecollectingduct(distal
nephron).
Proteinbasedcylindricalmoldsoftherenaltubule
Resultofdamagetotherenaltubule
Cancontaincellsandothermaterial
Dehydrationandacidicurineespeciallypredisposestocastformation.
Typesofcasts
Protein
RBC
WBC
Other
Urinemicroscopy
Urinarybacteria
Abundantnormalmicrobialfloraofthevaginaorexternalurethralmeatus.
Abilitytorapidlymultiplyinurinestandingroomtemperature.
However,itshouldbeinterpretinviewofclinicalsymptoms.
Bacterialculturemustbedoneincaseofbacteriuria
Urinemicroscopy
UrinaryYeast
Yeastcellsmaybecontaminantsorrepresentatrueyeastinfection.
Often difficult to distinguish from red cells and amorphous crystals
butcanbedistinguishedbytheirtendencytobud.
Mostoftentheyarecandida,whichmaycolonizebladder,urethraor
vagina.
Urinemicroscopy
Nonbacterialorganisms
General "crud" or unidentifiable objects may find their way into a

specimen,particularlythosethatpatientsbringfromhome.
Spermatozoacansometimesbeseen.
Rarely,pinwormovamaycontaminatetheurine.
ovafrombladderinfestationswithschistosomiasismaybeseen.
WBCsinurine WBCscasts WBCscasts
Mixedcasturine Cellularcasts Nonrenalepithelialcells
Urineoxalatecrystals WBCandRBCsinurine Sq.epitheliumcell

BacteriaandWBCs Mucous Buddingyeast
Hyalinecast Redbloodcellcast Whitebloodcellcast
coarselygranularcast waxycast Amorphousurates

Uricacidcrystals Calciumoxalatecrystals Amorphousphosphates
Triplephosphatecrystals Ammoniumuratecrystals Calciumphosphates
Calciumcarbonatecrystals Tyrosinecrystals Cystinecrystals

Lucinecrystals Cholesterolcrystals Sulfacrystals
Fecalmaterial Talcinurine Startchinurine
Cottonfibersinurine Cottonfibersinurine Epithelialcellsinurine

Requestingaurinalysis
Signsandsymptoms
Burning urination, difficulty in urination & painful urination

infectiousandobstruction.
Semen in urine with great exhaustion (semonorrhoea) prostate
problem.
Backache early in the morning infection, renal stones (fullnes of
kidney,pelvis).
UTI (leucorrhoea in females) spread of infection into UT
(honeymooncystitis).
Fatigueandexhaustionrenalproblems.
Lower abdominal pain or supra pubic pain due to infection or
stones.
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Biochemicalcompositionof

Nature Precedings : doi:10.1038/npre.2011.6595.1 : Posted 12 Nov 2011

Proteinmetabolism 2NH3+CO2H2NC0NH2 Urea

Nucleicacidmetabolism Majorportionfrompurines Uricacid

Random First Afternoon 24hours

Different colors of urine

Under 750 ml (oligouria) dehydration, infection, obstruction, renal

Over 2000ml (polyuria) diabetes insipidus, hypertension,

Specific gravity reflects kidney's ability to concentrate.

Determination of urinary osmolality is more important

The normal osmolality of random urine specimen varies from 401350

Reagent strips are used only once and discarded.

BRIEFLY dip the strip in urine.

Presence of glucose (glycosuria) indicates that the blood

Bilirubin is a byproduct of the breakdown of hemoglobin.

Ketones are excreted when the body metabolizes fats

Presence of blood may indicate infection, trauma to the

pH measures degree of acidity or alkalinity of urine

Presence of protein (proteinuria) is an important indicator

Urobilinogen is a degradation product of bilirubin formed

Nitrite formed by gram negative bacteria converting

Leukocytes (white blood cells) usually indicate infection.

Negative results for glucose, ketones, bilirubin, nitrites,

Handling and Storage

Lighting Observe color changes and color charts

QC Reagent strips should be tested with positive

Sample Proper collection and storage of urine is

Testing cold specimens would result in a slowing

Inadequate mixing of specimen could result in false

Renal tubular epithelial cells, usually larger than

Positive nephrotic syndrome, condition leading to

General "crud" or unidentifiable objects may find their way into a

WBCsinurine WBCscasts WBCscasts

Mixedcasturine Cellularcasts Nonrenalepithelialcells

Urineoxalatecrystals WBCandRBCsinurine Sq.epitheliumcell

BacteriaandWBCs Mucous Buddingyeast

Hyalinecast Redbloodcellcast Whitebloodcellcast

coarselygranularcast waxycast Amorphousurates

Uricacidcrystals Calciumoxalatecrystals Amorphousphosphates

Triplephosphatecrystals Ammoniumuratecrystals Calciumphosphates

Calciumcarbonatecrystals Tyrosinecrystals Cystinecrystals

Lucinecrystals Cholesterolcrystals Sulfacrystals

Fecalmaterial Talcinurine Startchinurine

Cottonfibersinurine Cottonfibersinurine Epithelialcellsinurine

Burning urination, difficulty in urination & painful urination

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