Johll Urinalysis Lab

8/20/2019 Johll Urinalysis Lab

1/15

Experiment 8

Urine and Blood Analysis

Instructor Notes and Answers

Part A Qualitative Analysis of Blood and Urine Electrolytes

o Please review all Material Safety ata S!eets for t!e c!emicals used in t!is

la"oratory#

o Prepare all standard simulated "lood solutions from $ % of distilled water to

w!ic! one drop of red food colorin& !as "een added# Use t!is solution as t!e

"lan' simulated "lood#

o Prepare all standard simulated urine solutions from $ % of distilled water to

w!ic! one drop of yellow food colorin& !as "een added# Use t!is solution as

t!e "lan' simulated urine#

o (ou may c!oose to distri"ute your standards and "lan's in vials or test tu"es

at t!e "e&innin& of t!e la"# Alternatively) you may c!oose to !ave t!ese

solutions at a central location and !ave t!e students o"tain t!eir samples as

needed t!rou&!out t!e la" session# *ompara"ly) students will need more "loodand urine "lan' in order to perform a "lan' test for eac! analysis#

+est for Sodium and Potassium Ions

o Prepare t!e "lood and urine sodium standards to approximately ,#$ M# +!is is

close to t!e concentrations of real samples# (ou may use any sodium salt -e#)sodium c!loride) sodium nitrate.# Students s!ould see a "ri&!t oran&e flame for "ot! t!e "lood and urine

standards prior to the wire glowing # /it! t!e "lan's) t!e students s!ould

only see an oran&e flame w!en t!e wire "e&ins to &low#

o Prepare t!e "lood and urine potassium standards to approximately ,#$ M# +!is

is close to t!e normal concentration in a real urine sample) "ut 0,12,3 &reatert!an a real "lood sample# Students s!ould see a pale violet flame for "ot! t!e "lood and urine

standards prior to t!e wire &lowin Students may see an oran&e flame as

t!e wire "e&ins to &low# /it! t!e "lan's) t!e students s!ould only see anoran&e flame w!en t!e wire "e&ins to &low#

+est for *!loride

o Prepare t!e "lood c!loride sample to approximately ,#$ M in "lood "lan'#

+!is is also close to t!e concentration of a real sample# If you used sodium

$


2/15

c!loride w!en preparin& t!e "lood sodium standard) you may use t!e same

solution# Students s!ould see t!e appearance of a w!ite A&*l precipitate wit! t!e

addition of t!e silver nitrate solution#

o Prepare t!e silver nitrate solution to ,#,2 M in distilled water#

+est for P!osp!ate

o Prepare t!e "lood p!osp!ate standard solution to ,#$ M in "lood "lan'# +!is is

very !i&! compared to a real "lood sample -i#e#) ,#,,$ M.# (ou may use p!osp!ate or !ydro&en p!osp!ate salts# Sodium p!osp!ate wor's well#

Students s!ould see t!e solution turn yellow after !eatin& several minutes

in t!e water "at! followin& t!e addition of t!e nitric acid and ammonium

moly"date# +!e yellow color is due to a precipitate t!at eventually falls outof solution#

+est for *alcium

o Prepare t!e "lood calcium standard solution to ,#$ M in "lood "lan'# *alcium

c!loride wor's well# +!is is very !i&! compared to a real "lood sample -i#e#)

,#,,4 M.# Students s!ould see t!e appearance of a w!ite calcium oxalate precipitate

wit! t!e addition of t!e ammonium oxalate solution# If students !ave difficulty seein& t!e precipitate) t!e p5 may "e too low

-increasin& t!e solu"ility of calcium oxalate. and few drops of sodium!ydroxide or ammonium !ydroxide may "e added#

o Prepare t!e ammonium oxalate solution to ,#$ M in distilled water#

+est for Ammonium

o Prepare t!e "lood ammonium standard solution to ,#$ M in "lood "lan'# +!is

is very !i&! compared wit! a real "lood sample -i#e#) ,#,,,2 M.# Students s!ould see t!e wetted red litmus paper turn "lue as ammonia

vapor leaves t!e solution#

Part B Qualitative Analysis of Urine Proteins) 6lucose) and 7etones

+est for Proteins

o Prepare t!e urine protein standard solution wit! $#, m% of e&& w!ite to every

09: m% of "lan' simulated urine# If t!e standard "ecomes muc! more

concentrated t!an t!is) t!e solution will !ave a distinct cloudy appearance# Students s!ould "e a"le to see t!e foamin& of t!e proteins as t!ey "ecome

denatured in t!e !eat and t!e solution "e&ins to "oil#

0


3/15

+est for 6lucose

o Prepare t!e simulated urine &lucose standard wit! $ & &lucose to every 2, m%

"lan' simulated urine#

After t!e addition of t!e Benedict;s solution and !eatin&) students s!ould "e a"le to see a "ric' red precipitate "e&in to form and eventually settle att!e "ottom of t!e test tu"e#

o Prepare $,, m% of Benedicts solution wit! $#250=) $, & sodium

car"onate) $


4/15

7idney ailure) Blood? potassium c!loride and potassium p!osp!ate

P!osp!ate concentration will "e twice w!at was tested in t!e

standards) "ut t!is will not affect t!e analysis# ia"etesrelated) Urine? e&& w!ites) acetone) &lucose) sodium c!loride

ia"etesrelated) Blood? sodium nitrate or any ot!er sodium salt except

p!osp!ate or c!loride# 5eart ailure) Urine? e&& w!ite

5eart ailure) Blood? any ammonium salt except c!loride or p!osp!ate

Pre&nancyrelated) Urine? (ou may c!oose to add &lucose or a sodium salt

to any urine un'nown to su&&est pre&nancy# Pre&nancyrelated) Blood? NA

o isclaimer? +a"le 8#$ su&&ests si&nificantly !i&! levels of species in urine and

"lood t!at indicate a medical conditionC !owever) t!is experiment is tailored to

a moc' environment and s!ould not "e used for !ealt!related dia&noses of

real people#

9


5/15

Experiment 8 Urine and Blood Analysis Answers

$# +a"le 8#$ s!ows si&nificantly !i&! levels of species -e#) sodium. tested in t!isexperiment correspondin& wit! potential !ealt! conditions# Assume t!at t!e tests

you performed in t!is experiment only &ave positive results -i#e#) formation of

color or precipitate. w!en t!e concentration of t!e species was &reater t!anDnormal levels# Usin& +a"le 8#$ and your results for your un'nown samples

-ta'en from t!e deceased woman.) determine if t!e woman may !ave potentially

died from one of t!e listed conditions# Also determine if s!e could !ave "een pre&nant# Explain your reasonin

Answers will vary "ased on t!e un'nowns supplied# Be sure to ma'e sure t!estudents explain t!eir reasonin

0# Usin& your c!emistry text or t!e Internet) explain w!y t!e color of t!e flame is

different for potassium and sodium standard samples# 5int? (ou will need to

"riefly discuss atomic structure) ener&y levels) and electrons#

+!e atomic structure of eac! atom is different) resultin& in a distri"ution ofelectronic ener&y levels uni@ue for eac! element# +!is distri"ution presents

various ener&y level electron transitions# +!e flame excites t!e atom forcin&

an electron from a lower ener&y level to a !i&!er ener&y level# /!en t!e atomrelaxes) a p!oton of li&!t is &iven off correspondin& to t!e uni@ue ener&y

difference for potassium or sodium#

4# Usin& your c!emistry text or t!e Internet) identify t!e precipitate products for t!ec!loride and calcium tests#

Silver c!loride and calcium oxalate

2


6/15

Experiment 8

Urine and Blood Analysis

Purpose

+o introduce t!e student to "lood and urine analysis tec!ni@ues of common electrolytes)

proteins) &lucose) and 'etones# Students will "ecome familiar wit! common precipitatin&

and colorc!an&in& reactions#

Time Duration 0 1 4 !ours

Materials

="tain t!e followin& materials?$# Standard simulated "lood and urine samples of potassium and sodium

0# Standard simulated "lood samples of c!loride) calcium) ammonium) and

p!osp!ate4# Blan' simulated urine and "lood samples9# $, test tu"es

2# +est tu"e clamp

F# Bunsen "urner


7/15

Nurses and doctors commonly re@uest a "lood andHor urine sample in an attempt to

dia&nose !ealt! conditions) w!ile toxicolo&ists re&ularly analye "lood and urine to aid

medical examiners in t!e determination of cause of deat!#+!e composition of "ot! "lood and urine c!an&es as a function of !ealt!# Urine is

often analyed for t!e presence of proteins) &lucose) andHor 'etones# +!ese species are not

present to any apprecia"le de&ree in t!e urine of a !ealt!y individual# Presence of one ormore of t!ese species at si&nificant levels may indicate de!ydration) 'idney dama&e) or

dia"etes# In addition) urine and "lood are "ot! fre@uently analyed for electrolyte levels

of sodium and potassium) w!ile "lood is additionally analyed for t!e presence ofc!loride) p!osp!ate) calcium) and ammonium# All of t!ese electrolytes are endo&enous to

"ot! urine and "lood and !ave defined normal levels# 5owever) an increase or decrease in

t!eir normal levels may indicate a condition of !eart failure) dia"etes) or starvation#

+a"le 8#$ s!ows si&nificantly !i&! levels of electrolytes) proteins) &lucose) and'etones -as indicated "y an Dx. in urine and "lood of six potentially fatal medical

conditions# Elevated levels for pre&nancy are also s!own# (ou will use +a"le 8#$) in

conJunction wit! your results from t!is experiment of "ot! simulated "lood and urine

samples ta'en from t!e corpse of an unidentified female) to presumptively determinecause of deat!# In addition) you will determine w!et!er or not s!e may !ave "een

pre&nant#

Table 8.1. Elevated %evels in Urine -U. and Blood -B. as a unction of Medical

*onditions

Protein-U.

7etones-U.

6lucose-U.

Sodium-U.

Potassium-U.

Sodium-B.

Potassium-B.

*!loride-B.

P!osp!ate-B.

*alcium-B.

Ammonium-B.

e!ydration K K K K K

Starvation K K

5eavy Metal Poisonin& K

7idney ailure K K K K K K

ia"etesrelated K K K K K

5eart ailure K K

Pre&nancyrelated K K

Procedures

Part A Qualitative Analysis of Blood and Urine Electrolytes

$# ="tain standard simulated "lood and urine samples of potassium and sodium#

="tain standard simulated "lood samples of c!loride) calcium) ammonium) and

p!osp!ate# Also o"tain one simulated "lood and one simulated urine sample'nown not to contain any of t!e a"ove species -"lan' samples.# %a"el all of your

samples#

+est for Sodium and Potassium Ions?

0# ="tain approximately 8 inc!es of nic!rome wire# Bend a small loop at one end to

allow trappin& of a small amount of li@uid sample#


8/15

4# Prepare and li&!t your Bunsen "urner# ip t!e loop of your nic!rome wire into

your "lan' simulated "lood sample# Introduce t!e wire loop into t!e flame and

record your results#LL (ou will "e performin& a flame test# +!e color of t!e flame prior to t!e wire

&lowin& oran&e will "e representative of t!e species present -i#e#) potassium or

sodium.# (ou may wis! to record t!e appearance of t!e flame "ot! prior to andafter t!e wire "e&ins to &low#

9# Gepeat step 4 wit! your standard simulated "lood sodium sample#

2# Gepeat steps 4 and 9 wit! your "lan' and standard simulated urine sodium

samples) respectively#

F# Gepeat steps 412 usin& your standard simulated "lood and urine potassium

samples#


9/15

$4# Add 412 drops of 4 M nitric acid and 0, drops of ammonium moly"date solution

to eac! test tu"e#

$9# Place t!e test tu"es upri&!t in t!e warmed water# Allow t!em to sit for 21$,

minutes# ecord your o"servations#

$2# +urn off your !ot plate or Bunsen "urner# ispose of your samples in an

appropriate waste container and rinse your test tu"es clean# (ou may need to use a

test tu"e "rus! for cleanin

+est for *alcium Ion?

$F# ispense $ m% of standard simulated "lood calcium sample into a test tu"e and$ m% of "lan' simulated "lood into a separate test tu"e#

$


10/15

04# +urn off your Bunsen "urner# ispose of your samples in an appropriate waste

container and rinse your test tu"es clean#

Part B Qualitative Analysis of Urine Proteins) 6lucose) and 7etones

$# ="tain standard simulated urine samples of protein) &lucose) and 'etones# %a"el

your samples if t!is !as not already "een done#

+est for Proteins?

0# ispense approximately $10 m% of standard simulated urine protein sample and

$10 m% of "lan' simulated urine sample into two separate test tu"es#

4# Prepare and li&!t your Bunsen "urner# 5old t!e urine protein test tu"e over t!e

flame of t!e Bunsen "urner# ollow all precautions &iven in part A w!en !eatin& a

test tu"e in an open flame# Gemove t!e test tu"e from t!e flame Just as it "e&ins to "oil# +!e precipitate formed in t!is experiment appears as Dsoapy "u""les or

frot! on t!e inside of t!e test tu"e# ecord your o"servations#

9# Gepeat step 4 wit! your "lan' simulated urine sample#

2# +urn off your Bunsen "urner# ispose of your samples in an appropriate waste

container and rinse your test tu"es clean#

+est for 6lucose?

F# Set up an apparatus similar to t!at in part A) step $$C !owever) t!e water will need

to "e&in "oilin& for t!is procedure#


11/15

$$# In a fume !ood) slowly add approximately $ m% of concentrated ammonium

!ydroxide to eac! test tu"e) so as to form a separate layer on top of t!e solutions#

$0# After $10 minutes) record your o"servations focusin& on t!e interface "etween

t!e two layers#

$4# ispose of your samples in an appropriate waste container and rinse your test

tu"es clean#

Part * Analysis of Un'nown Blood and Urine Samples

$# ="tain an un'nown simulated "lood and urine sample# ecord your un'nownnum"ers#

0# +est your un'nown simulated "lood and urine samples wit! procedures in parts A

and B# (ou do not need to run "lan' samples unless re@uired "y your instructor#ecord all of your o"servations#

4# etermine in eac! case if t!e species "ein& tested is present# Explain your

reasonin

9# *lean up w!en finis!ed#

$$


12/15

Name? ate?

Experiment 8

Urine and Blood Analysis /or's!eet

esults and !bser"ations

Part A #ualitati"e Analysis o$ Blood and Urine Electrolytes

o ="servations

Standard Sample Blan' Sample

Blood sodium

Urine sodium

Blood potassium

Urine potassium

Blood c!loride

Blood p!osp!ate

Blood calcium

Bloodammonium

$0


13/15

Part B #ualitati"e Analysis o$ Urine Proteins% &lucose% and 'etones

o ="servations

Standard Sample Blan' Sample

Urine proteins

Urine &lucose

Urine 'etones

Part ( Analysis o$ Un)nown Blood and Urine *amples

o ="servations Un'nown Os

Un'nown Sample Present or not present Explain#

Blood sodium

Urine sodium

Blood potassium

Urine potassium

Blood c!loride

$4


14/15

Un'nown Sample Present or not present Explain#

Blood p!osp!ate

Blood calcium

Blood

ammonium

Urine proteins

Urine &lucose

Urine 'etones

$9


15/15

Name? ate?

Experiment 8 Urine and Blood Analysis #uestions

9# +a"le 8#$ s!ows si&nificantly !i&! levels of species -e#) sodium. tested in t!is

experiment correspondin& wit! potential !ealt! conditions# Assume t!at t!e tests

you performed in t!is experiment only &ave positive results -i#e#) formation of

color or precipitate. w!en t!e concentration of t!e species was &reater t!anDnormal levels# Usin& +a"le 8#$ and your results for your un'nown samples

-ta'en from t!e deceased woman.) determine if t!e woman may !ave potentially

died from one of t!e listed conditions# Also determine if s!e could !ave "een pre&nant# Explain your reasonin

2# Usin& your c!emistry text or t!e Internet) explain w!y t!e color of t!e flame is

different for potassium and sodium standard samples# 5int? (ou will need to

"riefly discuss atomic structure) ener&y levels) and electrons#

F# Usin& your c!emistry text or t!e Internet) identify t!e precipitate products for t!e

c!loride and calcium tests#

$2

Documents

Johll Urinalysis Lab