Labmanual Physics

Laboratory Manual for Physiological
Chemistry
Spring 2009
Table of Contents
Handling Emergencies...............................................................................2
Safety Regulations.....................................................................................3
General Laboratory Procedures..................................................................3
ntroduction to !rganic Com"ounds...........................................................3
solation of Chloro"hyll and Carotenoid Pigments from S"inach.................3
Chemical Pro"erties of #li"hatic and #romatic #lcohols.............................3
!$idation and Structure of Carbonyl Com"ounds......................................3
!"tical somers..........................................................................................3
Carbohydrates...........................................................................................3
#cid%&ase Reactions 'ith Carbo$ylic #cids and Esters...............................3
Synthesis of #s"irin...................................................................................3
Synthesis ( "ro"erties of Soa"..................................................................3
solation and Characteri)ation of Casein from Mil*.....................................3
#mylase+ ,he #cti-ity of an En)yme..........................................................3
nteraction of ./ Light 'ith Matter............................................................3
0
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1
Laboratory Rules
and Guidelines
Handling Emergencies
While we will do everything possible to ensure a safe environment,
accidents can occur. In case of the following emergencies, always inform
the instructor and do the following+
Burns % 2lush 'ith cool ta" 'ater.
Chemicals in the eye % C#LL 2!R HELP. 2orce the eye o"en3 4ush 'ith
'ater for at least 20 minutes.
Chemicals on the skin % 2lush 'ith 'ater. Rinse acid s"ill 'ith sodium
hydrogen carbonate solution5 bases 'ith boric acid.
Chemicals on your clothes %REM!/E 6!.R CL!,HES. 6ou do not 'ant
the chemicals to reach your s*in.
Clothing on fre % S,!P%7R!P%R!LL. .se a 8re blan*et or sho'er only if
you are standing 'ithin arm9s reach.
Cut in skin % Rinse immediately 'ith 'ater. ns"ect for glass. Get
medical attention.
2
Safety Regulations
1: Acceptable eye protection must be worn at all times ;earing contact lenses
in the lab is strongly discouraged and may be forbidden by your instructor.
2: Bare feet and sandals are not allowed in the lab S"illed chemicals and
bro*en glass on the 4oor can result in serious in<uries.
3: Shorts and short skirts are not allowed =either midri> nor shoulders may be
e$"osed 'hether standing straight3 reaching3 or bending o-er.
?: Each student must know the location of the safety e!uipment" 8re
e$tinguishers3 eye 'ashes3 safety sho'er and the e$it.
@: Horseplay and#or carelessness are prohibited
A: $o unauthori%ed e&periments are to be performed
B: ;or* is "ermitted only at the assigned time unless other'ise authori)ed by the
instructor. n any case3 $E'ER work alone.
C: Chemicals and eDui"ment are not to be remo-ed from the laboratory.
E: (o not sit on bench tops.
10: Eating) drinking) smoking or chewing anything is not permitted in the
laboratory
11: $e*er pipet by mouth5 al'ays use a "i"et bulb.
12: Be cautious when testing for odors. #l'ays 'a-e fumes to'ards your nose
'ith your hand. =E/ER smell a chemical directly.
13: Always add acid to the water or base3 ne-er do the o""osite.
1?: $e*er aim the opening of a test tube or +ask at yourself or anyone else
1@: $e*er lea*e reactions unattended if they in-ol-e heating or ra"id reactions.
1A: !nly the lab manual and lab noteboo* should be at lab counter. Book bags)
outerwear) etc are to be placed out,of,the,way in the location indication by
your instructor.
1B: Report any in-ury3 ho'e-er minor3 to the instructor at once.
1C: Always use tongs to handle hot ob-ects
1E: .oose clothing must not be 'orn in the laboratory. !"en s'eatersFhoodies3 loose
slee-es3 e$cessi-ely G4o'yH blouses.
20: Hair below chin must be tied back3 'hile 'or*ing in the laboratory.
21: Broken glass will be disposed of in the glass disposal bo&es3 not in the
regular trash.
22: (ispose of waste properly) as directed by your instructor.
23: Clean up all spills immediately
2?: Always check glassware for chips and cracks
reali)e that these R.LES ;LL &E E=2!RCE7 for my safety and for the safety of my lab
"artners and that failure to obser-e these rules3 in addition to resulting in unacce"table
safety ha)ards and the loss of 'or*ing time3 'ill result in e$"ulsion from the laboratory.
ac*no'ledge that ha-e recei-ed a 'ritten co"y of these regulations and ha-e been
gi-en the o""ortunity to discuss them 'ith the instructor.
=#ME IIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIC!.RSEIIIIIIISEC,!=IIIII
SG=#,.REIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIII7#,EIIIIIIIIIIIIIII
3
/eneral .aboratory 0rocedures
,he follo'ing "rocedures are intended to "re-ent contamination of chemicals3 and to
"romote safety3 smooth laboratory o"eration and laboratory eJciency.
Read labels
carefully
Some chemicals ha-e -ery similar names. n the case of acids and bases3 do
not assume that the reagent bottles are in the correct "laces.
Eyedropper
s#0ipets
=e-er use an eye dro""er or "i"ette in a reagent bottle unless there is a
dro""er 'ith the bottle for use only 'ith that bottle. 6our seemingly clean
eyedro""er may actually be dirty3 and end u" contaminating an entire bottle
of reagent. f you need to dis"ense a chemical 'ith an eyedro""er3 "our a
small amount into a bea*er and use the eyedro""er from there.
1nused or
e&cess
reagents
=e-er "our them bac* into a reagent bottle. Contamination of the contents
may result. 7is"ose of the e$tra "ro"erly. #long 'ith this3 do not ta*e large
amounts of reagents3 since e$cess amounts must be 'asted. 6ou can al'ays
get more if you need it.
2btaining
solids
!btain solids using a s"atula s"eci8ed for the "articular reagent3 or by
"ouring the solid out by rotating the bottle bac* and forth until the solid
'or*s its 'ay out of the bottle or -ial.
Cleaning
E!uipment
&e sure your eDui"ment is clean before use3 and be sure to return borro'ed
eDui"ment either as clean or cleaner than you found it. ;ash glass'are 'ith
soa" and ta" 'ater. Rinse 'ith ta" 'ater. !ccasionally3 some chemicals and
glass'are mar*ers must be remo-ed 'ith acetone. 7o a 8nal rinse 'ith a
small amount of distilled 'ater.
1sing
litmus and
other test
papers
#l'ays do litmus "a"er or "H "a"er tests by "lacing the test "a"er on a clean
'atch glass and transferring a dro" of the test solution to the litmus using a
stirring rod.
'ials and
3ars
#l'ays reca" -ials) <ars and bottles immediately after use. 7! =!, LE#/E
,HEM .=C#PPE7. Contamination of the contents may result3 s"illage could
occur if the container is *noc*ed o-er3 and many solid chemicals absorb
'ater from the air 'hich ma*es them ca*e or become stic*y ma*ing them
un'eighable.
1sing
Balances
,he balances are e$"ensi-e "ieces of eDui"ment and must be treated 'ith
res"ect. Please obser-e the follo'ing+
a. 7o not mo-e the balance from its s"ot unless s"eci8cally as*ed to do so.
b. Place ob<ects on the "an gently.
c. =e-er "lace chemicals directly on the "an. #l'ays use a container.
Clean s"ills on balances immediately. #s* your instructor for assistance.
4aste
(isposal
.se designated 'aste container to dis"ose 'aste after the e$"eriment.
?
.aboratory
$otebooks
# laboratory noteboo* is the most basic "iece of eDui"ment used in the laboratory. t
is -ery necessary to de-elo" a "ro"er method of using a noteboo*3 no matter 'hat science
one "ursues. t is one of the goals of this laboratory e$"erience to de-elo" good noteboo*
techniDue. E-eryone has his or her o'n style of recording data3 and di>erent instructors
may demand di>erent things. ,he guidelines described here are to gi-e you an idea of the
minimum that is ty"ically acce"table.
6our laboratory noteboo* is for recording data and obser-ations 'hile "erforming
e$"eriments. =otes from class discussion may a""ear in the noteboo*3 but they must be
clearly distinguished from e$"erimental data. Ksee belo':
,he "ur"ose of a laboratory noteboo* is to "ro-ide 6!. !R #=6!=E ELSE 'ith an
accurate record of 'hat 6!. 77 in the laboratory3 not necessarily 'hat you 'ere su""osed
to do. t should contain+
1: ,itle of the "ro<ectFe$"eriment "erformed
2: &rief descri"tions of 'hat you did.
2: Lualitati-e obser-ations.
3: =umerical data 'hich is 'ell%labeled as to 'hat it is3 and 'hich has the "ro"er units
of measurement.
?: nformation of any reference material used for the 'or*
#LL data and obser-ations are to be 'ritten in your noteboo*3 =!, in the margins of
reference materials3 handouts3 or laboratory manuals3 and =!, on any other sheets of
"a"er in the laboratory. 6ou 'ill not co"y o-er anything into another noteboo* at any time.
6our noteboo*s 'ill be handed in from time to time for grading. Grading 'ill be based in
"art on your adherence to these guidelines.
Concerning K1: abo-e3 you should 'rite do'n 'hat you did 'ithout co"ying 'hat the
instructions said. 2ragmentary sentences are 8ne3 as long as they are clear. f you as* a
Duestion of your lab instructor3 he or she 'ill "robably as* to see your noteboo* to see
'hat you did. 7! =!, "resent the instructions. ,hat says 'hat you should ha-e done3 not
'hat you did.
Remember...
You or another person should be able to use your notebook and
any sources you cite, and exactly reproduce what YOU DID in the
laboratory.
/uidelines
1. 4hile in the lab write 2$.5 in your notebook. 7! =!, 'rite on scra" "a"er or
anything else. 7! =!, transcribe your noteboo* in any form. ,he original is the only
@
legitimate co"y. 6ou may 'rite in your noteboo* outside of lab as long as you "ro"erly
date the entries and 7! =!, CH#=GE #=6,H=G ,H#, ;#S PRE/!.SL6 E=,ERE7.
2. 4rite what you did ,his may be di>erent from 'hat you 'ere su""osed to do. f you
ha-e a record of 'hat you did3 you may ha-e a 'ay of 8guring out 'hat 'ent 'rong if
something does go 'rong.
3. The frst two pages of a notebook should be reser*ed for a Table of Contents)
'hich is continually u"dated. Sometimes noteboo*s already ha-e a table of contents
section in them.
?. The pages must be numbered starting at 1 for the 8rst "age and continuing on each
"age Keach side of the sheet: until the end of the noteboo*. =oteboo* "ages are
numbered <ust li*e boo* "ages are. Some noteboo*s come 'ith the "ages
"renumbered.
@. The date 6including the month) day and year7 must appear on each page of
the notebook ,he date must also a""ear 'hene-er data is being ta*en on a ne'
date.
A. The name of the e&periment must be at the beginning of the notes for
that e&periment ,his is usually3 but not reDuired to be3 the name of the e$ercise. t
could be your o'n title for 'hat is being done.
B. ,he source of the e$"eriment3 that is3 the full bibliographic citation of 'here your
"rocedures are ta*en from3 must appear at the beginning of an e&periment #t
any time if another source is used3 it must also be "ro"erly cited. f any data are ta*en
from reference or other boo*s3 these must be "ro"erly cited. 6our instructor 'ill gi-e
you an e$am"le.
C. $otebooks must be written in blue or black pen) =!, in "encil. .se only
standard blue or blac* in* "lease.
E. $2 erasures should e-er be made. #lso3 no 4hite,out is to be used. f you 'rite
something incorrectly3 sim"ly dra' a S=GLE line through it and continue to 'rite. #lso3
you should $E'ER o*erwrite anything. #gain3 sim"ly "utting a single line through
the error and re'riting is the best "olicy.
10.0ages must ne*er be torn out of a notebook ,he original "ages must remain
intact.
11.E*erything must be labeled. ,hat is3 each "rocedure to be "erformed must be
clearly identi8ed. #ll data and obser-ations must be clearly labeled as to 'hat it is3
'hat the units of measurement are3 etc. 2or e$am"le3 labeling the mass of an ob<ect3
say a test tube3 as Mtest tube 2.0 gM is insuJcient. MMass of test tube 2.0 gM 'ould ha-e
a better label. #l'ays label your data so that se-eral days after recording it you 'ill be
able to *no' 'hat it is. &e liberal 'ith headings and subheadings. Headings such as
MPart 1M and MPart 2M are not suJcient. ;hat are you doing in Part 1N 6ou must also
e$"licitly distinguish bet'een e$"erimental data and obser-ations3 notes from grou"
discussions3 and your o'n conclusions or hy"otheses.
12.f you use an instrument such as a balance or spectrophotometer) always include
the brand name and model number Kif a-ailable:. #lso3 many of our instruments
are numbered3 for e$am"le MS"ectronic 20 O@M. ,his number should be included. ,his is
es"ecially im"ortant if you disco-er at a later time that the data you recorded do not
ma*e sense. Perha"s there 'as an instrument malfunction. f you *no' s"eci8cally
'hich instrument you used3 that "ossibility can be chec*ed out.
13.6our noteboo* does not ha-e to be so neat and orderly that it is ready to be "ublished
in M=oteboo*s &eautifulM3 but it should be suJciently organi)ed and legible so that you
or someone else could use it to reproduce your experiment or write a report.
89 (o not use your lab notebook for any other course
1@.There should be only one notebook) no other copy
A

:ntroduction to
2rganic Compounds
/oals for the Student"
Learn to identify organic functional grou"s
Learn to classify organic com"ounds based u"on their functional grou"s
Learn to name organic com"ounds based u"on their functional grou"s
Construct models of al*anes to -ie' three%dimensional structure of al*anes.
n-estigate the relationshi" bet'een a structural formula and a three dimensional
molecule using molecular models
Construct models of isomers of al*anes 'ith the same molecular formula.
dentify isomers3 structural formula3 condensed structural formula and s*eletal
formulas.
:ntroduction
n this e$ercise 'e 'ill be introduced to organic com"ounds. !rganic chemistry is the
study of com"ounds that are "rimarily com"osed of carbon and hydrogen atoms. !ther
"rominent elements in organic chemistry are o$ygen3 !3 nitrogen3 =3 sulfur3 S3 and the
halogens K4uorine3 23 chlorine3 Cl3 bromine3 &r3 and iodine3 :. Since all organic com"ounds
contain some amount of carbon and hydrogen atoms3 organic com"ounds are identi8ed
and classi8ed by the functional grou"s they "ossess. # functional group is a grou" of
atoms that react in a "redictable 'ay. Com"ounds 'ith the same functional grou" are
classi8ed into a "articular class of organic com"ounds and the name is deri-ed from
belonging to that class.
;unctional /roups
Table 8" 2rganic ;unctional /roups
;unctional /roup Class Characteristic E&ample
C C
#l*ane !nly carbon%carbon
single bonds
H
3
C CH
3
C C
#l*ene Carbon%Carbon double
bond
H
2
C CH
2
C C
#l*yne Carbon%carbon tri"le
bond
HC CH
B
#romatic Si$ atom carbon ring
'ith alternating
double and single
bonds
H
H
H
H
H
H
X
X = F, Cl, Br, or I
Haloal*ane
!ne or more halogen
atoms
H
3
C Cl
OH
#lcohol Hydro$yl grou" K%!H:
H
3
C OH
O
Ether !$ygen atom bonded
to t'o carbon atoms
H
3
C O CH
3
SH
,hiol # PSH grou" bonded
to a carbon atom
H
3
C SH
C
O
H
#ldehyde Carbonyl grou"
Kcarbon%o$ygen
double bond: 'ith PH
H
3
C C
O
H
C
O
Qetone
Carbonyl grou"
bet'een t'o carbon
atoms
H
3
C C
O
CH
3
C
O
O H
Carbo$ylic
#cid
Carbo$yl grou"
Kcarbon%o$ygen
double bond and P!H:
H
3
C C
O
O H
C
O
O
Ester
Carbo$yl grou" 'ith P
H re"laced by a
carbon
H
3
C C
O
O CH
3
N
#mine
=itrogen atom 'ith
one or more carbon
grou"s
H
3
C NH
2
C
C
O
N
#mide Carbonyl grou"
bonded to a nitrogen
atom
H
3
C C
O
NH
2
$omenclature
=omenclature of organic com"ound is go-erned by the nternational .nion of Pure and
#""lied Chemistry K.P#C: system. ,he system is founded on t'o ma<or "rinci"les5 K1:
determine the longest continuous carbon chain and K2: number3 name3 and al"habeti)e all
substituents. ,hese "rinci"les ha-e been elaborated into the follo'ing set of nomenclature
rules.
8 Straight chain compounds with only carbon and hydrogen atoms 6Alkanes7
a. count the number of carbon atoms
b. add the ending ane to the "re8$ corres"onding to the correct number of
carbon atoms.
Table <" :10AC $ames for the ;irst Ten Continuous,Chain Alkanes
=umber of Carbon #toms Pre8$ =ame Molecular 2ormula
1 meth methane CH?
2 eth ethane C2HA
3 "ro" "ro"ane C3HC
? but butane C?H10
@ "ent "entane C@H12
A he$ he$ane CAH1?
B he"t he"tanes CBH1A
C oct octane CCH1C
E non nonane CEH20
10 dec decane C10H22
< Alkanes with substituents
a. 'rite the name of the longest continuous chain of carbon atoms
b. number the carbon atoms starting from the end nearest the 8rst substituent
to generate the lo'est set of numbers
c. gi-e the location and name of each substituent as a "re8$ to the al*ane
name
i. "lace a hy"hen bet'een the number and the substituent name
ii. al"habeti)e the substituents
iii. use a "re8$ Kdi%3 tri%3 tetra%3 etc: if a substituent a""ears more than
once and use commas to se"arate t'o or more numbers.
Table =" Common Substituent $ames and Structures
Substituent Structure $ame
H
3
C
methyl
H
3
C CH
2
ethyl
H
3
C CH
2
CH
2
"ro"yl
E
C
H
3
C
H
3
C
H
iso"ro"yl
H
3
C CH
2
CH
2
CH
2
butyl
C CH
2
H
3
C
H
3
C
H
isobutyl
C
CH
3
CH
3
H
3
C
tert%butyl K
t
butyl:
X
KR S 23 Cl3 &r3 or :
4uoro3 chloro3 bromo3 iodo
H
3
C CH
Cl
CH
2
CH
CH
3
CH
2
CH
3
1 2 3 4 5 6
2%chloro%?%methylhe$ane
H
3
C C
Cl
CH
2
CH
CH
3
CH CH
3
6 5 4 3 2 1
Cl
Br
2%bromo%@3@%dichloro%3%methylhe$ane
= Cycloalkanes
a. count the carbon atoms in the ring and add the "re8$ cyclo to straight chain
name
b. substituent rules from abo-e a""ly e$ce"t the 8rst substituent is al'ays
"laced on carbon 1
c. al"habeti)e to determine substituent on carbon 1
CH
2
CH
3
H
3
C
1%ethyl%3%methylcyclo"entane
9 Alkenes and Alkynes
a. name the longest continuous carbon chain that contains the double or tri"le
bond
i. re"lace the ane ending of the al*ane 'ith ene for an al*ene and
yne for an al*yne
10
b. number the longest continuous carbon chain from the end nearest the
double or tri"le bond
c. gi-e the location and name of each substituent as a "re8$ to the al*ene or
al*yne name
i. "lace a hy"hen bet'een the number and the substituent name
ii. al"habeti)e the substituents
iii. use a "re8$ Kdi%3 tri%3 tetra%3 etc: if a substituent a""ears more than
once and use commas to se"arate t'o or more numbers.
1 2 3 4 5
H
2
C CH CH CH
CH
3
CH
3
?%methyl%2%"entene
HC C CH
1 2 3 4 5
CH
Br
CH
3
Cl
3%bromo%3%chloro%1%"entyne
> Aromatics
a. monosubstituted ben)ene rings are named as ben)ene deri-ati-es using the
substituent name
Table 9" Common ?onosubstited Aromatic Compuonds
Structure .P#C =ame Common =ame
CH
3
methylben)ene toluene
NH
2
ben)eneamine aniline
OH
hydro$yben)ene "henol
b. disubstituted ben)ene rings are numbered to gi-e the lo'est number to the
substituents
i. common "re8$es are often used K132 substitution is ortho3 133
substitution is meta3 and 13? substitution is "ara:
c. if a ben)ene ring is a substituent Klongest chain is more than si$ carbons or
contains a double or tri"le bond:3 then it is named a "henyl grou"
so"ro"ylben)ene
11
Cl
Cl
132%dichloroben)ene or
orthochloroben)ene
H
3
C CH
2
CH CH
2
CH
2
CH
2
CH
3
1 2 3 4 5 6 7
3%"henylhe"tane
A. Alcohols @ Thiols
a. name the longest continuous carbon chain containing the hydro$yl grou" K%
!H:
i. re"lace the ane ending of the al*ane 'ith ol ending
b. number the longest continuous carbon chain starting at the end closest to
the hydro$yl grou"
c. name and number other substituents relati-e to the hydro$yl grou"
d. name a cyclic alcohol as a cycloal*anol 'ith all cycloal*ane rules a""lying for
substituents
e. a""ly aromatic naming rules for ben)ene rings containing a hydro$yl grou".
,he base name is then "henol
f. thiols are named by adding thiol to the al*ane name of the longest
continuous carbon chain bonded to the PSH grou"
i. the location of the PSH grou" is indicated by numbering the main
chain from the closest end
1 2 3 4
H
3
C CH
OH
CH
CH
3
CH
3
3%methyl%2%butanol
OH I
?%iodo%3%cyclohe$anol
H
3
C CH
SH
CH
2
CH
CH
3
CH
2
CH
3
1 2 3 4 5 6
?%methyl%2%he$anethiol
A Ethers
a. 'rite the al*ane name of the larger al*yl grou" as the main chain
b. name the o$ygen and smaller al*yl grou" as a substituent called an alo!y
grou"
H
2
C O CH
2
H
3
C CH
2
CH
3
Etho$y"ro"ane
O CH
3
metho$yben)ene
12
B Aldehydes @ Cetones
a. for an aldedyde3 name the longest continuous carbon chain containing the
carbonyl grou" by re"lacing the e in the al*ane name 'ith al
i. name and number any substituents on the carbon chain by counting
the carbonyl carbon as carbon 1
b. for a *etone3 name the longest continuous carbon chain containing the
carbonyl grou" by re"lacing the e in the al*ane name 'ith one
i. number the main chain starting from the end nearest the carbonyl
grou"
ii. name and number any substituents on the carbon chain
H
3
C CH
2
CH
2
C H
O
4 3 2 1
butanal
H
2
C CH CH
2
C H
O
5 4 3 2 1
H
3
C
CH
3
3%methyl"entanal
H
2
C C CH
2
CH
3
5 4 3 2 1
H
3
C
O
3%"entanone
H
2
C CH
2
6 5 4 3 2 1
H
3
C C
O
CH
Cl
CH
3
2%chloro%3%he$anone
D Carbo&ylic Acids
a. name the longest continuous carbon chain containing the carbonyl grou"
and re"lace e of the al*ane name 'ith oic acid
b. number the carbon chain beginning 'ith the carbo$yl grou" as carbon 1
c. gi-e the location and names of substituents on the main chain
d. for the aromatic ben)oic acid3 number the ring from the carbo$yl grou" as
carbon 1
H
2
C CH
2
H
3
C C
O
OH
4 3 2 1
butanoic acid
C CH
2
H
2
C C
O
OH H
3
C
CH
3
CH
3
5 4 3 2 1
333%dimethyl"entanoic acid
13
8EEsters
a. 'rite the name of the carbon chain from the alcohol as an alyl group
b. 'rite the name of the carbo$ylic grou" as carbo!ylate 'ith an oate ending
H
3
C CH
2
O C
O
CH
2
CH
2
CH
3
ethyl "ro"anoate
H
3
C O C
O
methyl ben)oate
88Amines @ Amides
a. for amines3 name the longest continuous carbon chain bonded to the
nitrogen atom and re"lace the e in the al*ane name 'ith amine
i. number the carbon chain to sho' the "osition of the amine grou" and
any other substituents
ii. in secondary and tertiary amines3 use the "re8$ "# to name smaller
al*yl grou" attached to the = atom.
b. amides are named by dro""ing the oic acid from the carbo$ylic acid name
and adding amide
H
3
C CH
2
CH
2
NH
2
Pro"anamine
H
3
C CH
2
CH
2
NH
CH
3
=%methyl "ro"anamine
H
2
C CH
2
H
3
C C
O
NH
2
butanamide
H
2
C CH
2
H
3
C C
O
N
CH
3
CH
3
=3=%dimethyl butanamide
#lso3 in this e$ercise 'e 'ill study the three dimensional structure of some al*anes3
using a molecular model *it to reinforce the nomenclature for al*anes and some of theirTs
deri-ati-es. n each ty"e of al*ane each carbon has four -alence electrons and must
al'ays ha-e four single bonds to other carbon3 hydrogen or halogen atoms. ,he bond
arrangement of four single bonds used by carbon in al*ane is sho'n as belo'.
&onding Pattern of
Carbon
#rrangement of &onds around
Carbon
S"atial Structure and &ond
#ngles
C

,etrahedr
al
1?
,o understand the three dimensional structure of organic com"ounds3 models can be
build using a ball and stic* model *it. n this *it3 there are colored s"heres 'hich re"resent
the atoms drilled to recei-e connecting bonds. 7i>erent color s"heres3 blac* for carbon
and red for o$ygen3 are used to re"resents di>erent *inds of atoms and a color code for
atoms 'ill be included in the model *it. Each of the s"heres Katoms: has the correct
number of holes for bonds K'ooden or "lastic stic*: that attach to other s"heres.
a: Kb: (c)
,he three dimensional structure of the al*ane models re"resents -ery closely resembles
the a""ro$imate geometry Ksha"e and angle: of the molecules they re"resent. ,'o
structures are identical if they are superimposable%that is3 if one structure can be "lace
Gon to"H of another so that all colored s"heres coincide. Methane is the 8rst member of
al*anes and three di>erent structure of methane is sho'n abo-e3 'hich re"resents the
structural formula Ka:3 three dimensional structures Kb: and ball and stic* model Kc:.
Com"ounds ha-ing the same molecular formula can be re"resented by more than one
structure and each structure includes the same grou" of atoms but a di>erent s"atial
arrangement of the atoms. ,hese com"ounds are called isomers. somers ha-e the same
molecular formula but di>erent three dimentional structures. !ne structure cannot be
con-erted to the other 'ithout brea*ing and forming ne' bonds.
,he isomers ha-e di>erent "hysical and chemical "ro"erties. !ne of the reasons for the
-ast array of organic com"ounds is the "henomenon of isomerism. Many biological
reactions are -ery s"eci8c and in-ol-e only one isomer.
somers of C?H10
CH
3
CH
2
CH
2
CH
3

CH
3
CH
CH
3
CH
3
somers of C2HA!
H C C
H
H
H
H
O H

C O C
H
H
H
H
H
H
n%butane 2%methyl
"ro"ane
ethyl alcohol dimethyl ether
E&perimental
,his is a t'o 'ee* e$ercise. n the 8rst 'ee* you should com"lete and turn into your
instructor the nomenclature re"ort sheet. 7uring the second 'ee* you 'ill "erform the
structure "ortion of the e$"eriment3 com"leting and turning in the second re"ort sheet.
1@
,he model *it includes di>erent colored s"heres re"resenting di>erent atoms and grey
connectors for re"resenting bonds. Carbon atoms are blac* s"heres and ha-e four holes
that re"resent the four co-alent bonds that carbon atoms al'ays form. Hydrogen atoms
are 'hite s"heres and only form one bond. ,he green s"here re"resents chlorine atom and
o$ygen atoms are red s"here. Halogen atoms form one bond and o$ygen atoms form t'o
bonds5 s"heres for these atoms 'ill ha-e the a""ro"riate number of holes.
Single co-alent bonds are re"resented by grey connectors3 'hich insert into holes of
the atoms. ,o conser-e time and de"ending u"on the number of "ieces in your model *it3
you may use only the stic* to re"resent the C% H bonding arrangement.
6ou 'ill be 'or*ing in grou"sKt'oFthree: to construct the models of di>erent
com"oundsKal*ane3 haloal*ane3 haloalcohol: using the model *its. Each model must be
in-estigated for geometry Ksha"e ( angle:
1A
$age intentionally blan.
1B
RE02RT SHEET,:ntroduction to 2rganic Compounds 64eek
87
$ameFFFFFFFFFFFFFFFFFFFFFFFFFFFF 0artnerGname FFFFFFFFFFFFFFFFFFFFFFFFF
SectionFFFFFFFFFFFFFF (ateFFFFFFFFF
$ame the following compounds
: Alkanes
Br
IIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIII IIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIII
:: Alkenes # Alkynes
IIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIII IIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIII
::: Aromatics
1C
:' Alcohols @ Thiols
OH
HS
' Ethers
O
O
': Aldehydes @ Cetones
H
O
Br
O
':: Carbo&ylic Acids
HO
O
HO
O I
I I
'::: Esters
1E
O
O O
O
:H Amines @ Amides
NH
2
N
O
Page intentionally blan*.
20
RE02RT SHEET I :ntroduction to 2rganic Compounds
64eek <7
: Construct a model of methane) CH
9
21
a: ;hat is the geometry associated 'ith this structure N IIIIIIIIIIIIIIIIIIIIIIIIIIII
b: ;hat is the -alue of the H C H bond anglesN IIIIIIIIIIIIIIIIIIIIIIIIIIIII
:: Construct a model of chloromethane) CH
=
Cl
a: 7ra' a 'edge and hash mar* to re"resent the three dimensional sha"e of the molecule.
b: s the geometry the same as methaneN IIIIIIIIIIIIIIIIIII
c: #re the hydrogen atoms eDui-alent Ki.e.3 do they ha-e identical en-ironments 'ith res"ect to the
other atoms ad<acent to themsel-es:NIIIIIIIIIIIIIIIIII
::: Construct a model of chloromethanol) CH
<
Cl62H7
a: s the geometry the same as the "re-ious t'o structuresN IIIIIIIIIIIIIIIIIIII
b: #re the hydrogen atoms attached to the carbon atom eDui-alentN IIIIIIIIIIIIIIII
c: ;hat "art of the name indicates the alcohol grou"N IIIIIIIIIIIIIIIIIII
:' Construct a model of ethane) C
<
H
J
a: 7ra' a condensed structural formula of C2HA IIIIIIIIIIIIIIIIIIIIIIIIIIIIII
b: 7ra' a structural formula of C2HA
c: #re the t'o carbon atoms of C2HA eDui-alentN IIIIIIIIIIIIIIIIIIIIIIIIIII
d: #re the si$ hydrogen atoms of C2HA eDui-alentN IIIIIIIIIIIIIIIIIIIIIIIIIIII
' Construct a model for chloroethane) CH
=
CH
<
Cl
a: #re the carbon atoms in CH
3
CH
2
Cl eDui-alentN IIIIIIIIIIIIIIIIIIII
22
b: #re the hydrogen atoms in CH
3
CH
2
Cl eDui-alentN IIIIIIIIIIIIIIIIIIIII
': Construct all possible models for dichloroethanes) C
<
H
9
Cl
<
a: Ho' many structural isomer e$ist for C
2
H
?
Cl
2
N
I
IIIIIIIIIIIIIIIIIIIII
I
b: 7ra' condensed structural formulas for each structural isomers of C
2
H
?
Cl
2
.
':: Construct all possible models for propane) C
=
H
B
a: 7ra' a structural formula for C
3
H
C
and using sDuares3 triangles3 and For circles3 indicate
the carbon atoms that are eDui-alent to each other
b: #re the eight hydrogen atoms of C
3
H
C
are eDui-alentN IIIIIIIIIIIIIIII
c: s there a relationshi" bet'een eDui-alent carbons and eDui-alent hydrogensN f so3 state
the relationshi".
'::: Construct all possible models for chloropropane) C
=
H
A
Cl
a: Ho' many structural isomers e$ist for C
3
H
B
Cl N IIIIIIIIIIIIIIIIIIIIIIII
b: 7ra' condensed structural formula for each structural isomer of C
3
H
B
Cl
23
:H Construct all possible models for C
9
H
8E
KHint+ straight -ersus branched
chain:
?
H
10
N IIIIIIIIIIIIIIIIIIIIIIII
b: 7ra' condensed structural formulas for each structural isomers of C
?
H
10
. #lso3 using
sDuares3 triangles3 and For circles3 indicate the carbon atoms that are eDui-alent to each
other
2?
H 1sing your C
9
H
8E
models from abo*e) remo*e one hydrogen atom
and replace it with a chlorine atom to make diKerent structural
isomers of C
9
H
D
Cl
?
H
E
Cl.N IIIIIIIIIIIIIIIIIIIIIII
b: 7ra' condensed structural formulas for each structural isomer of C
?
H
E
Cl.
H: (raw line,bond formulas for all possible structural isomers of
C
9
H
B
Cl
<
) which are formed by replacing hydrogen atoms in the
*arious isomers of C
9
H
D
Cl

6e&ercise H b7 with a second chlorine
atom
2@
:solation of
Chlorophyll and
Carotenoid
0igments from
Spinach
#da"ted from+ Pa-ia3 7. L.5 Lam"man3 G. M.5 Qri) G. S.5 Engel3 R. G. Introduction to %rganic
&aboratory 'echni(ues) # Microscale #""roach 3
rd
Edition Saunders College Publishing+ =e'
6or*3 =63 1EEE and also Luach3 H. ,.5 Stee"er3 R. L..5 GriJn3 G. ;.3 *. +hem. ,duc.3 200?3 C13
3C@%3CB. ,oni &ell K200?:
Learn the techniDues of e$traction and "uri8cation of chemical com"ounds from
natural "roducts.
Learn the techniDue of identi8cation of di>erent com"onents in chemical
com"ounds isolated from
natural "roducts
:ntroduction
S"inach3 a green leafy -egetable usually can be gro'n as a s"ring and fall cro" in
the cooler =orth #merican climate. S"inach is a source of /itamin # and it is rich in iron3
and calcium. ,he lea-es contain a number of colored "igments3 generally falling into t'o
categories+ chlorophylls and carotenoids.
Carotenoids are "art of a larger collection of "lant deri-ed com"ounds called
terpenes. ,hese naturally occurring com"ounds contain 103 1@3 203 2@3 303 and ?0 carbon
atoms. :soprene is the basic 8-e%carbon building bloc* of the ter"ene class of biological
com"ounds. #lso *no'n as 2%methyl%133% butadiene3 these units are lin*ed in a Ghead to
tailH fashion to build the structure of ter"enes. ,'o iso"renes are lin*ed together to ma*e
one terpene unit. ,he branched end is the GheadH and the unbranched end is the GtailH.
Carotenoids are tetrater"enes Keight iso"rene units:.
S"inach lea-es contain chlorophyll a and b and L,carotene as 'ell smaller
amounts of other "igments such as &anthophylls 'hich are o$idi)ed -ersions of carotenes
and pheophytins 'hich loo* li*e chloro"hyll e$ce"t that the magnesium ion Mg
U2
has
been re"laced by t'o hydrogen ions H
U
. Chloro"hylls a and b are the "igments that ma*e
"lants loo* green. ,he double bonds are con-ugated3 meaning they occur bet'een e-ery
2A
Isoprene is the basic -ve#carbon
building bloc of the terpene class
of biological compounds.
head end
tail end
other "air of carbons3 and allo' ca"ture the Knongreen: light energy used in
"hotosynthesis.
V%Carotene is a carotenoid and it causes carrots and a"ricots to be orange. ;hen
ingested3 V%carotene is clea-ed to form t'o molecules of /itamin #. /itamin #3 also called
retinol3 "lays an im"ortant role in -ision and ser-es as an anti,o&idant.
2B
+hlorophyll a .left/ and
chlorophyll b .right/ are
very similar molecules. +an
you spot the di0erences1
'hese small changes are
enough to change their
color.
#+arotene .top/, #
carotene .middle/, and
!anthophylls .e!ample
on bottom/ have very
similar structures. +an
you spot the
di0erences1 'hese
small changes are
enough to change their
color. 2our body can
only use #carotene to
mae 3itamin 4. Why1
n this e$"eriment 'e 'ill isolate and use di>erences in "olarity of the "igments to
e>ect a se"aration. Chloro"hylls and carotenoids are slightly di>erent in "olarity. 7ue to
their lo-ely color3 'e 'ill easily follo' the se"aration -isually. V%Carotene is a hydrocarbon
and it is -ery non"olar. &oth chloro"hylls contain C ! and C = bonds 'hich are "olar
and also contain magnesium bonded to nitrogen 'hich is such "olar bond that it is almost
ionic. &oth chloro"hylls are much more "olar than V%Carotene. ,here is another structural
di>erence in bet'een the chloro"hyll a and b3 Chloro"hyll a has a methyl grou" K CH3: in a
"osition 'here chloro"hyll b has an aldehyde grou" K CH!:. ,his ma*es chloro"hyll b
slightly more "olar than chloro"hyll a.
Since s"inach also contains cellulose3 iron3 and 'ater soluble -itamins in addition to
chloro"hylls and carotenes3 'e ha-e to ha-e a method of se"arating all these com"ounds.
,he most common a""roach to isolating these bioacti-e natural "roducts is e&traction.
Chloro"hylls and carotenes are relati-ely non,polar organic substances com"ared to other
com"onents5 hence3 they are more soluble in organic sol-ents li*e dichloromethane or
acetone. Since Wli*e dissol-es li*eT3 these sol-ents 'ill be suitable to selecti-ely e$tract
these com"ounds into organic sol-ent and lea-e the other com"ounds behind. ,o do the
e$traction3 you 'ill 8rst grind u" the s"inach in a little bit of acetone. ,he green acetone
'ith s"inach com"ounds is called e&tract. .nfortunately acetone 'ill dissol-e almost
anything3 including the stu> that you do not 'ant.
Chloro"hylls and carotenes do not dissol-e -ery 'ell in 'ater5 they dissol-e li*e
cra)y in he$ane. ,he other com"ounds do not dissol-e 'ell in he$ane3 but dissol-e 'ell in
'ater. Li*e 'ater and oil3 'ater and he$ane are immiscible5 they Wdo not mi$T. ,he
he$ane 'ill form a layer on to" of the 'ater Kli*e oil does: because he$ane is less dense
than 'ater. #fter -igorous sha*ing to mi$ the layers tem"orarily3 you 'ill allo' them to
se"arate. ,he lo-ely green chloro"hylls and yello' carotenes 'ill lea-e the 'ater at the
bottom to dissol-e in the he$ane layer at the to". #fter "i"etting%o> he$ane layer3 the
di>erent com"onents in the "igment mi$ture 'ill be analy)ed by using thin layer
chromatogra"hy.
,he number of com"ounds in the he$ane e$tract can be Duic*ly determined by a
techniDue called thin layer chromatography3 'hich is abbre-iated GT.C.H 6ou 'ill "ut a
little s"ot of your e$tract on a "lastic "late coated 'ith silica gel. Silica gel is a -ery "olar
substance. ,he "lates are "laced in a container 'ith a mi$ture of sol-ents. n this case3 the
sol-ents 'ill be Duite non%"olar. ,he sol-ents 'ill begin to tra-el u" the "late3 li*e a 'ic*.
Some of the com"ounds in the he$ane e$tract 'ill be more "olar and 'ill stic* to the s"ot
on the silica. !ther com"ounds in the he$ane e$tract 'ill be less%"olar to di>ering degrees
and 'ill tra-el u" 'ith the sol-ent Since there are many le-els bet'een totally "olar and
totally non%"olar3 the com"ounds can be se"arated by "olarity. ,he more aJnity a
com"ound has for the sol-ent3 the farther u" the "late it 'ill tra-el.
7i>erent com"ounds should rise to di>erent heights on your ,LC "late5 ho'e-er the
e$act height a "articular com"ound rises de"ends on ho' high the sol-ent is allo'ed to
rise u" the "late. f the sol-ent tra-els higher3 then the s"ots all tra-el higher too. ,o
correct for this di>erence and generate a number 'hich can be com"ared to re"orted
-alues or to other indi-idualTs 'or*3 the retention fraction or Rf. -alue is calculated. ,he
retention fraction is de8ned to be the fractional rise of the s"ot com"ared to the rise of the
sol-ent. ,he Rf -alue for a com"ound 'ill change if a di>erent de-elo"ing sol-ent or a
di>erent ty"e of "late is used. #fter you ha-e de-elo"ed your ,LC "late 'ith your he$ane
e$tract you ha-e to calculate Rf -alues for each s"ot on your "late. S"ots 'ith the same Rf
-alues 'ithin e$"erimental error and the same a""earance should be the same com"ound.
2C
#n e$am"le3 if you had <ust t'o com"onents in your original e$tract3 this is 'hat your
results might loo* li*e+

E&perimental
E$traction of "igment from lea-es+
'his is a 3,52 colorful e!periment and you need to make note of the colors of things
at each step. 2ou should draw the centrifuge tube, for e!ample, and label the layers and
the colors. 6eep in mind that the term 7clear8 refers to transparency while 7colorless8
refers to absence of color. 2ou can have a clear green solution or an opa(ue green
solution. +onversely, you can have a clear colorless solution or an opa(ue colorless
solution.

1. ;eigh about 1.@ g fresh s"inach lea-es KdonTt use stems: and record the mass and
obser-ations about the color. ,ear the lea-es into confetti%si)ed "ieces and "lace
these "ieces along 'ith 0.@ g of anhydrous magnesium sulfate and 1.0 g of sand
into a mortar Kthe bo'l "art of the mortar and "estle:.
2. Grind 'ith a "estle until a light green "o'der is obtained Kabout @%10 minutes:.
3. ,ransfer the "o'der mi$ture into a 1@ mL "lastic centrifuge tube 'ith a ca".
?. .sing the sDuee)e bottle3 add roughly 1.0 mL of acetone to the mortar to rinse.
@. ,ransfer the rinse acetone to the centrifuge tube containing the "o'der 'ith a
0asteur pipette. 2our instructor will show you how to use a pipette properly as
part of your pre#lab discussion.
A. Re"eat ste"s ? and @.
B. f the -olume of acetone has e-a"orated to less than roughly 2.0 mL3 as measured
using the mar*ings on the centrifuge tube3 then add enough to ma*e%u" the
-olume.
C. Ca" and sha*e the mi$ture. &e sure to -ent the tube occasionally by pointing
away from you and others and loosening the ca". #llo' the tube to stand for a
fe' minutes so the solid material may se"arate.
E. ,ransfer the liDuid from centrifuge tube to a clean centrifuge tube 'ith a Pasteur
"i"ette. ;rite Ge$tractH on the second tube3 along 'ith your initials.
10. #dd about 2.0 mL of he$ane and 2.0 mL distilled 'ater to the e$tract. Ca" and
sha*e the mi$ture. &e sure to -ent the tube occasionally by pointing away from
you and others and loosening the ca". #llo' the tube to stand for a fe' minutes
so the layers may se"arate. dentify the he$ane layer and the 'ater layer KHo'
2E
can you do this if you donTt *no'N:. Ma*e a labeled s*etch of the tube3 contents3
and colors in your noteboo*.
11. Remo-e the 'ater layer 'ith a Pasteur "i"ette and transfer it to a small bea*er
labeled G'aste.H
12. #dd another 2.0 mL of distilled 'ater to the he$ane layer in the centrifuge tube as
a 'ash. Ca" and sha*e the mi$ture. &e sure to -ent the tube occasionally by
pointing away from you and others and loosening the ca". #llo' the tube to
stand for a fe' minutes so the layers may se"arate. Remo-e the 'ater layer 'ith
a Pasteur "i"ette and transfer it to a small bea*er labeled G'aste.H
13. #lthough 'ater and he$ane Wdo not mi$TXin reality a little bit of 'ater 'ill stay in
the he$ane. 6ou can tell there is 'ater in the he$ane layer if it is a little cloudy.
6ou must dry Kremo-e 'ater from: the he$ane layer by adding a drying agent
called anhydrous sodium sulfate K=a2S!?:. 2our instructor will show you how to
use a drying agent. # cou"le of micros"atula scoo"s is usually suJcient.
1?. #llo' the drying agent to settle and then transfer the he$ane to a small -ial 'ith a
ca". Label the -ial and then "roceed 'ith thin layer chromatogra"hy.
,hin Layer Chromatogra"hy+
1. Pre"are the T.C chamber by "lacing one half of a 8lter "a"er into the <ar. ,hen
"our the sol-ent mi$ture3 "ro-ided by instructor3 o-er the 8lter "a"er into the
bea*er until it is about 0.@ cm dee". Place the lid on the chamber. ,his does t'o
things+
#. it *ee"s all the sol-ent from e-a"orating
&. it allo's the air inside the chamber to become saturated 'ith sol-ent.
MM $ote" e*en slight changes in the composition or contamination of the
de*eloping sol*ent will lead to diKerences in Rf *alues (onGt let the
chamber sit for long periods of time between plates :f a chamber gets
contaminated) prepare a fresh oneMM
2. !btain a T.C plate and /ER6 lightly dra' a "encil line K=! =Q: about 1 cm from
the bottom. f you "ress too hard3 the silica gel 'ill come o>X in 'hich case you 'ill
to get a ne' ,LC "late. .sing a ca"illary tube3 ma*e a s"ot of your e$tract on the
"encil line. 6ou may ha-e to let the s"ot to dry and then s"ot it again if it isnTt dar*
enough carefully "lace the s"otted "late into the chamber and re"lace the lid.
MM $ote" if your spot goes under the sol*ent) it will not tra*el up the plate
0repare a new plate if this happensMM
3. 6ou 'ill immediately see the sol-ent start to tra-el u" the "late. ,he line of sol-ent
mo-ing u" is called the Wsol-ent frontT. !nce the sol-ent front is roughly 1 cm of the
to" of the "late3 remo-e the "late and Duic*ly mar* the sol-ent front 'ith a "encil.
?. #lthough most of the s"ots are easily -isuali)ed by the na*ed eye3 use the ./ lam"
to insure that you are noting all "ossible s"ots. Lightly circle each s"ot in "encil.
Lightly label each s"ot K#3 &3 C3 etc.:.
@. 7etermine Rf -alues for all of your s"ots. ,his 'ill gi-e you Duantitati-e -alues for
com"arison. Measure the distance from the starting line to the sol-ent front. ,hen3
measure to the center of each s"ot. 7i-ide the center s"ot distance by the sol-ent
front distance5 this is the Rf -alue. ,he higher the Rf -alue3 the less "olar the
com"ound
A. ,ry to match them to the com"ounds sho'n belo' Klisted in order of decreasing Rf
-alues:+
30
Carotenes K1%2 yello'%orange s"ots:
Pheo"hytin # Kgray intense:
Pheo"hytin & K gray3 may only be -isible under ./:
Chloro"hyll # Kblue%green3 intense:
Chloro"hyll & Kgreen:
Rantho"hylls Kas many 3 yello' s"ots:
31
RE02RT SHEET,:solation of 0igments from Spinach
$ameFFFFFFFFFFFFFFFFFFFFFFFFFFFF 0artnerGs name FFFFFFFFFFFFFFFFFFFFFFFFFF
1. Mass of s"inach IIIIIIIIIIIIIIIIII
2. ,o the right s*etch the layers in centrifuge tube
and clearly label the 'ater and he$ane layers.
3. ;hy must you occasionally -ent the tube during sha*ingN
? 7ra' a s*etch of your ,LC "late3 labeling the s"ots. ,hen3 8ll in the table
32
S"ot R
f
Probable dentity
,LC "late
@. ;ere there any s"ots your that does not match 'ith the "robable com"oundsN
A. ;hich com"ound is the most "olarN ;hich one is least "olarN E$"lain the reason for
your ans'er.
B. ;hat do you thin* 'ould ha""en if you used in* to mar* the s"otting lineN
33
Chemical
0roperties of
Aliphatic and
Aromatic Alcohols
Learn to identify the -isible obser-ations in a chemical reaction
Learn the di>erences in reacti-ity of 1
o
3 2
o
3 3
o
alcohols 'ith strong o$idi)ing reagent
Learn the di>erences in reacti-ity of ali"hatic and aromatic alcohol
Learn the chemical reactions in-ol-e con-erting one functional grou" into another
:ntroduction
Subclass General formula E$am"les
Primary

C OH
H
H
R
2-methylpropan-1-ol
Secondary

C OH
H
R'
R
butan-2-ol
,ertiary

C OH
R''
R'
R
2-methylpropan-2-ol
Phenol

C
C
C
C
C
C
H
H
H
H
H
OH

phenol
Alcohols3 aldehydes3 and ketones3 are three -ery im"ortant classes of o$ygen
containing organic com"ounds. #lcohols are classi8ed into primary K1
o
:3 secondary K2
o
:
3?
and tertiary K3
o
: according to the "resence of substituents in the carbon containing the
hydro&yl grou". 0henol is a class of aromatic com"ounds containing a hydro$yl grou"
attached to a ben%ene ring. ,hree subclasses of alcohols and "henol are sho'n on the
"receding "age.
n 8rst "art of this e$"eriment you 'ill learn the di>erence bet'een 1
o
3 2
o
3 and 3
o

alcohols and aromatic alcohols 'ith res"ect to their reacti-ity 'ith the strong o$idi)ing
reagent sodium dichromate. ,his can be easily demonstrated by noting a color change
'hen the Cr in the UA o$idation state of the orange colored dichromate ion3 Cr2!B
2%
3 is
reduced to the green colored chromium K:3 Cr
3U
3 ion. Simultaneously3 an a""ro"riate
alcohol is o$idi)ed to either an aldehyde Kand subseDuently to a carbo&ylic acid: or a
ketone. !f course3 if there is a no redo$ reaction there 'ill be no obser-ed color change.
n general the follo'ing unbalanced reaction describes the redo$ reaction
#lcohol U Cr2!B
2%
Carbo$ylic acid Kor *etone:
U Cr
3U
Kcolorless: Korange: Kcolorless: Kgreen:
2or the remaining "arts of the e$"eriment3 the chemical "ro"erties of ali"hatic alcohols 'ill
be e$amined. Here 'e 'ill com"are the solubility3 acidFbase "ro"erties and the reacti-ity
'ith iron K: chloride Kferric chloride: of a similar si)ed ali"hatic alcohol 'ith that of the
aromatic alcohol K"henol:.
E&perimental
!$idation 'ith #cidic 7ichromate
1. !btain Y @ mL of acidic dichromate solution K"re"ared "re-iously by mi$ing 3 mL of
a @Z sodium dichromate and 1 mL of concentrated sulfuric acid: and "lace Y 1 mL
KY20 dro"s: into four se"arate3 clean3 dry small test tubes and note the color.
Caution" the dichromate solution can potentially burn your skin or make
holes in your clothes f you s"ill any of this reagents re"ort the s"ill immediately
to your instructor so that it may be cleaned u" in an a""ro"riate manner.
2. ,o the se"arate test tubes add Y 0.@ mL KY10 dro"s: of ethyl alcohol3 iso"ro"yl
alcohol3 t%butyl alcohol3 or aDueous "henol. Mi$ by 8nger 4ic*ing the test tubes.
3. !bser-e and record the colors of the resultant solutions. 9int) 4 table of results
may help you -nd things (uicly during a lab (ui:.
?. 7is"ose of the solutions in the s"ecial 'aste container for dichromate 'aste.
Solubility
1. .sing a s"atula or force"s3 "lace a "ea si)ed amount of solid "henol crystals into
t'o se"arate3 clean3 dry small test tubes.
2. ,o one add 1 mL KY20 dro"s: of distilled 'ater and to the other add 1 mL KY20
dro"s: of 3M =a!H Ksodium hydro$ide:. S'irl the test tubes eDually and note the
relati-e s"eed 'ith 'hich the crystals dissol-e. Record your obser-ations.
3. Re"eat this "rocedure using 1 mL K20 dro"s: of cyclo"entanol instead of the "henol.
n this case you are adding a liDuid to a liDuid3 thus solubility is noted by a single
homogenous solution and insolubility by t'o layers.
?. Record your obser-ation and dis"ose of these solutions as indicated by your
instructor.
3@
#cidF&ase Pro"erties
1. Place 1 mL KY20 dro"s: of distilled 'ater3 ethyl alcohol and aDueous "henol into
three se"arate3 clean3 dry test tubes.
2. ,o each sam"le add one dro" of .ni-ersal ndicator solution and obser-e the color.
Com"are the solutions to the reference .ni-ersal ndicator color card and estimate
the solutionTs "H.
3. Record your obser-ations and dis"ose of these solutions as indicated by your
instructor.
Reacti-ity 'ith ron K: chloride K2eCl
3
:
1. !btain and obser-e the color of the iron K: chloride solution. Place 1 mL KY20
dro"s: of distilled 'ater3 cyclo"entanol and aDueous "henol into three se"arate3
clean3 dry test tubes.
2. ,o each sam"le add one dro" of the iron K: chloride solution. Mi$ by 8nger 4ic*ing
the test tubes.
3. !bser-e and record the colors of the resultant solutions and dis"ose of these
solutions as indicated by your instructor.
3A
3B
RE02RT SHEET,Chemical 0roperties of Alcohols
!$idation 'ith #cidic 7ichromate
1. a. 7ra' condensed structural formulas of ethyl alcohol3 iso"ro"yl alcohol3 t%butyl
alcohol3 and "henol.
b. Classify each of the "receding alcohols as 1[3 2[3 3[3 or aromatic.
2. ;hat is the function of the acid solution of sodium dichromateN
3. a. ;hat did you obser-e 'hen the sodium dichromate solution 'as added to ethyl
alcoholN
b. ;hat did you obser-e 'hen sodium dichromate solution 'as added to iso"ro"yl
alcoholN
c. ;hat did you obser-e 'hen the sodium dichromate solution 'as added to t%butyl
alcoholN
3C
d. ;hat did you obser-e 'hen the sodium dichromate solution 'as added to the
"henol solutionN
?. 7ra' condensed structural formulas for the organic "roducts of the abo-e reactions
that occur. f no reaction occurs3 'rite =R.
Solubility
1. a. 7id the "henol crystals dissol-e better in 'ater or in the =a!H solutionN
b. 7id the cyclo"entanol dissol-e better in 'ater or in the =a!H solutionN
c. &ased on the di>erent beha-iors of "henol and cyclo"entanol3 'hat
generali)ations can you ma*e about the solubility of similar si)ed ali"hatic and
aromatic alcoholsN
d. ;rite a full chemical eDuation for the chemical reaction that occurred 'hen =a!H
'as added to "henol.
e. ;rite a net ionic eDuation for chemical reaction that occurred 'hen =a!H 'as
added to "henol.
3E
#cidF&ase Pro"erties
1. &ased on the .ni-ersal ndicator Color Chart3 'hat is the a""ro$imate "H of+
a. ,he distilled 'aterN
b. ,he ethyl alcoholN
c. ,he aDueous "henolN
2. ;rite a full chemical eDuation for the chemical reaction that occurs 'hen "henol is
"laced in 'ater that e$"lains the obser-ed "H. s "henol an acid or a baseN
Reacti-ity 'ith ron K: chloride K2eCl
3
:
1. Can ferric chloride be used to distinguish aromatic alcohols from ali"hatic alcoholsN
E$"lain.
?0
?1
2&idation and
Structure of
Carbonyl
Compounds
Learn about the families of re"resentati-e carbonyl com"ounds
Study the beha-ior of re"resentati-e carbonyl com"ounds to'ard o$idi)ing agents
Learn about the di>erent functional grou"s in carbonyl com"ounds and in o$ygen
containing organic com"ounds
:ntroduction
n organic chemistry3 a carbonyl grou" is a functional grou" com"osed of a carbon
atom double bonded to an o$ygen atom+ C !. ,here are se-eral ty"es of carbonyl
com"ounds3 de"ending u"on 'hat is attached to the carbon atom in C !. ,he aldehyde
grou" is often 'ritten as CH!3 the *etone grou" is 'ritten as
CO
and the carbo&ylic
acid grou" is 'ritten as C!!H3 ester grou" 'ritten as C!!R and the amide grou"
'ritten as C!=H2. # carbonyl3 grou" characteri)es the follo'ing ty"es of common
com"ounds3 'here C! denotes a C ! carbonyl grou".
Compound Aldehyde Ketoe C!r"o#yl$% !%$d &'ter A($de
Structure
General
formula
)CHO )CO)* )COOH )COO)* )CONH)*
,he aldehyde grou" occurs in molecules of most sugars3 li*e glucose. ,he *etone
grou" is occurs also in one common sugar3 fructose. ,he amide grou" occurs in all amino
acids3 the building bloc* of "rotein.
?2
Carbonyl com"ounds are -ery reacti-e due to the di>erence in electronegati-ity
bet'een the carbon and the o$ygen atom. !$ygen is more electronegati-e than carbon3
and thus "ulls electron density a'ay from carbon to increase the bondTs "olarity. ,he
o$ygen is said to carry a partial negati*e charge or Gdelta minusH and 'ill be attracted
to "ositi-e s"ecies in solution5 for e$am"le3 a "roton in an acidi8ed solution or the carbon of
another carbonyl. ,he o$ygen is a nucleophile. ,he carbon is said to carry a partial
positi*e charge or Gdelta "lusH and 'ill be attracted to negati-e s"ecies in solution5 for
e$am"le3 the o$ygen of an alcohol or 'ater. ,herefore3 the carbonyl carbon is an
electrophile.
n this e$"eriment 'e are going to loo* at both the structure and the o$idation of
some carbonyl com"ounds. n the 8rst "art the action of mild o$idi)ing agent 'ill be
e$amined. n the ,ollenTs ,est3 'e 'ill use an o$idi)ing agent called the G,ollenTs
Reagent.H ,ollenTs Reagent contains sil-er diamine ion \#gK=H3:2]
U
'hich can o$idi)e
some categories of carbonyl%containg com"ounds to carbo$ylic acids. ,he #g
U
is
reduced to metallic sil-er "roducing a sil-er mirror on the glass'are. ,he ,ollenTs test
reaction is sho'n in the follo'ing generic e$am"le+
RCH!
KaD:
U 2#gK=H
3
:
2
]
U
KaD:
U 3!H
%
KaD:
RC!!
%
KaD:
U ?=H
3KaD:
U 2#g
Ks:
U 2
H
2
!
n the second "art of the e$"eriment ball and stic* models of o$ygen containing
com"ounds including carbonyl com"ounds distributed in the laboratory 'ill be
e$amined. 2rom the models you 'ill determine 'hat functional grou" family Kalcohols3
ethers3 aldehydes3 *etones3 hemiacetals3 or acetals: it belongs to and dra' its structure.
E&perimental
,ollenTs ,est
1. !btain four small test tubes and clean thoroughly 'ith detergent and a brush. Rinse
'ell 'ith ta" 'ater and 8nally 'ith distilled 'ater5 sha*e out e$cess 'ater. ,o each add
1 mL K20 dro"s: of @Z sil-er nitrate solution and 1 dro" of 3M =a!H3 mi$ 'ell.

2. ,o each tube add 2Z ammonium hydro$ide dro" by dro" until the grey sil-er o$ide
K#g2!: <ust dissol-es forming the soluble #gK=H3:2
U
ion solution. &e careful not to add
e$cess ammonium hydro$ide as this decreases the sensiti-ity of the test.
3. ,o the 8rst tube add 2 dro"s of 10Z glucose3 to the second tube add 2 dro"s of
formaldehyde3 to the third tube add 2 dro"s of acetone3 and to the fourth tube add 2
dro"s of iso"ro"yl alcohol5 be sure to label each test tube so that you *no' 'hich is
'hich. Record your obser-ations for each tube. Has a reaction already occurredN
?3
In every carbonyl, the more
electronegative o!ygen atom pulls
electron density away from the
carbon atom. 'he o!ygen is said to
be 7delta minus8 and will be
attracted to positive species in
solution. 'he carbon is said to be
7delta plus8 and will be attracted to
negative species in solution.
?. Mi$ the contents of each tube and "lace them into a hot 'ater bath. #fter se-eral
minutes remo-e the tubes and record your obser-ations.

@. Pour the contents of the test tubes into a 'aste container designated by your instructor.
Clean the tubes 'ith detergent and a brush. ,o remo-e any sil-er adhering to the test
tubes3 add small of concentrated nitric acid Kcaution conc. H=!3 can burn s*in and
clothes:. #dd contents to the 'aste container and clean the tubes.
Structures of Carbonyl Com"ounds
Some ball and stic* models of carbonyl com"ounds belonging to the families of aldehydes3
*etones3 carbo$ylic acids3 ester and other o$ygen containing com"ounds3 such as alcohol3
acetal3 hemiacetal and ether 'ill be distributed in this e$"eriment. 2or each model dra' a
line%bond formula3 and gi-e its functional grou" name Ki.e.3 alcohol3 *etone3 hemiacetal3
etc.:. .sing your results from the 8rst "art of the e$"eriment3 "redict if the com"ound
'ould react in the ,ollenTs ,est.
,he im"ortant "ieces are+
,he white sphere re"resents hydrogen atom
,he black sphere 'ith four holes re"resents carbon atom
,he red sphere re"resents o$ygen atom
,he gray sticks are for connecting carbon atoms to one another and to connect the
carbon atoms to hydrogen and o$ygen atoms
# stic* Kbond: attached to a carbon atom or an o$ygen atom and not connected to
anything else 'ill re"resent the C H or ! H bonding arrangement.
??
?@
RE02RT SHEET,2&idation and Structure of Carbonyl
Compounds
,ollenTs ,est
1. Record your obser-ations for the ,ollenTs test+
Glucose+
2ormaldehyde+
#cetone+
so"ro"yl alcohol+
2. 7ra' the structures and gi-e the names of the com"ounds that ga-e a "ositi-e
,ollenTs ,est.
3. Circle the functional grou" in the abo-e structures that is res"onsible for the
"ositi-e ,ollenTs test.
?A
?. ;hat is the name of this functional grou"N
Structur
e O
Line%bond 2ormula 2unctional
Grou"Ks:
Should React in
,ollenTs ,estN
1
2
3
?
@
A
?B
B
C
E
10
?C
?E
,o in-estigate the use of three dimensional structures to identify di>erent ty"es of
isomers
,o learn about stereoisomers3 and o"tical isomers
:ntroduction
Stereoisomers are isomers that ha-e same molecular and structural formulas but
di>erent s"atial arrangement. !"tical isomerism is one form of stereoisomerism. 2ptical
isomers are named li*e this because of their e>ect on "lane "olari)ed light.
#ll o"tical isomers contain a carbon atom <oined to four di>erent grou"s. ,he carbon
atom of this isomers are called Gasymmetric carbon atom or chiral centerH and the
molecule is called chiral.
!nly chiral molecules ha-e o"tical isomers. Some e$am"les of o"tical isomers are sho'n
belo'. ,he chiral center is mar*ed 'ith a star.

&utane%2%ol

2-Hydroxy propionic acid
Or Lactic acid

2-Aminopropionic acid
or Alanine
# carbon atom 'ith the four di>erent grou"s attached 'hich causes this lac* of
symmetry is described as a chiral center or as an asymmetric carbon atom. f you
cannot 8nd a "lane of symmetry the molecule is chiral. Practice on the molecules on the
ne$t "age. 7%alanine3 in a 'edge%and%dash formula belo'%right3 does not ha*e a plane
of symmetry and is chiral. Glycine on the left belo' has a plane of symmetry and
is thus achiral.
@0
2ptical :somers
Glycine%"lane of
symmetry

#lanine%no "lane of
symmetry
Chemist de-elo"ed methods to facilitate the -isuali)ation of 3%dimensional s"atial
arrangements of atoms or grou"s of atoms in a 2%dimensional en-ironment3 i.e.3 the "lane
of this "a"er. ,he most common method for "resenting 3%dimensional structures in a 2%
dimensional "lane is the ;ischer pro-ection. 2ischer "ro<ections are formed 'hen the
obser-er orients a tetrahedral structure such that the atoms or grou" of atoms in the
-ertical "lane are a'ay from the obser-er Kdashed 'edges: 'hile atoms or grou"s of
atoms in the hori)ontal "lane are to'ards the obser-er Ksolid 'edges:.
;hene-er a 2ischer "ro<ection is seen it
is meant to re"resent the orientation of atoms
or grou"s of atoms attached to the central
tetrahedral carbon atom3 as sho'n in the
8gure at the left. t is im"ortant to mention that
the cross in a 2ischer "ro<ection re"resents
chiral carbon.
.sing 2ischer "ro<ection formula3 3%
dimensional information of any molecule can
be oriented in 2%dimensional surface. Stereoisomers can be classi8ed into se-eral
di>erent ty"es of isomers. Enantiomers are stereoisomers that are t'o
nonsu"erim"osable com"lete mirror images of each other much as oneTs left and right
hands are the same but o""osite. Enantiomers ha-e similar "hysical and chemical
"ro"erties e$ce"t for their ability to rotate the "lane of "olari)ed light in the same amount
but in o""osite direction. ,his "ro"erty is called o"tical acti-ity
!ther ty"es of stereoisomers are diastereomers3 'hich are t'o
nonsu"erim"osable non mirror images3 and mesoisomers3 'hich contain more than one
chiral carbon atom but are o"tically inacti-e because of the symmetry of the molecule5
the mirror images of these com"ounds are su"erim"osable ,y"ically a meso com"ound
@1
can be identi8ed by noting that itTs 2ischer "ro<ection has a mirror "lane3 i.e.3 the to" and
the bottom hal-es of 2ischer "ro<ection are mirror images of each other. 7iastereomers of
glyceraldehydes of 233% dichlorobutane are sho'n belo'.

CH
3
CH
3
H Cl
H Cl
CH
3
CH
3
Cl H
Cl H
CH
3
CH
3
H Cl
Cl H
CH
3
CH
3
Cl H
H Cl
Enantiomer Kchiral: Meso Kdentical3 achiral:
E&perimental
Construct the 3%dimensional model for the follo'ing molecules using the
ball and stic* model *it containing blac* s"here as carbon atom3 'hite
s"here as hydrogen atom red s"here as o$ygen atom and green s"here as
nitrogen atom. ;ae the notes in your laboratory noteboo.
: &uild the structure of 233%dihydro$ybutanal K7% and L%
glyceraldehyde:.
a: 7ra' the 2ischer "ro<ection of molecule 'ith aldehyde grou"s
are on the to" of the structures.
b: Label the t'o isomers 'ith the ty"e of stereoisomers3 these structures re"resent.
:: &uild the structure of the four stereoisomers for 2333?3%
trihydro$ybutanal K7% and L% threose and 7% and L%
erythrose:.
a: 7ra' the 2isher "ro<ection of the structures 'ith aldehyde
grou"s are at the to" of the structure and hydrogens and
hydro$yl grou"s attached to chiral carbon s "oint to'ards
you.
b: .sing 2ischer "ro<ections for 7% and L% glyceraldehyde reference com"ound3 label
each of these 2ischer "ro<ections using 3e.g.3 713 L13etc.dentify the relationshi"s
bet'een -arious "airs of models.
@2
::: &uild the structure of all stereoisomers for
#s"aragine.
a: 7ra' 2ischer "ro<ections of all stereoisomers 'ith
carbo$ylic acid at the to" of the structure.
b: Label the isomers as 7% or L% as"aragine 'ith reference to
the 7% or L% glyceraldehyde structure. .se carbo$ylic
acid and the amino grou" of as"aragin as analogous to
the aldehyde and hydro$yl grou"s of the reference com"ound.
:' &uild the structures of all of stereoisomers for ,artaric acid.
a: 7etermine ho' many structure you can build and dra' the
corres"onding 2ischer "ro<ections for all the structures
'ith a star mar* for each chiral center.
b: Circle the structure that 'ould not be o"tically acti-e
' &uild the structure for 2333?3@3A% "entahydro$yhe$anal Kall are isomers
of glucose:
a: Construct the model so that the aldehyde grou" is at the to" of
the molecules and the hydro$yl grou" on the last chiral carbon
Kfurthest from the aldehyde grou": is "ointing to the right3 thus
generating a 7% structure for an aldohe$ose sugar. #lso build
your model such that the hydrogens and hydro$yl grou"s on each
chiral carbon are oriented to'ards you.
b: 7ra' the 2ischer "ro<ection corres"onding to your model.
7esignate each chiral carbon 'ith an asteris* ^. 7etermine ho'
many chiral center in the 'hole molecule. =otice the molecule3
'hether it formed chain or coiled structure and ho' the last
hydro$yl and the carbo$ylic grou" are "ositioned in the model. Predict the grou"
formed and the structure of the molecule if the hydro$yl and the aldehyde grou" react
together.
dentify eight di>erent stereoisomers can be formed 'ith 7%form3 one of 'hich is the
-ery im"ortant monosaccharide3 7%Glucose. Sho' your model and 2ischer "ro<ection
to instructor. Com"are your 2ischer "ro<ection 'ith eight "ossible 7%he$ose to
determine 'hich sugar structure you ha-e.

@3
RE02RT SHEET,2ptical :somers
: 7ra' and label the 2ischer "ro<ections for 233%dihydro$ybutanal.
:: 7ra' and label the 2ischer "ro<ections for 2333?%trihydro$ybutanal.
a: Ho' many chiral centers are thereN IIIIIIIIIIIIIIIIIIIIIIIIIIIII
b: Ho' many total stereoisomers are thereN IIIIIIIIIIIIIIIIIIIIIIIIIIII
c: Ho' many "airs of enantiomers are thereN IIIIIIIIIIIIIIIIIIIIIIIIIII
d: Ho' many "airs of diastereoisomers are thereN IIIIIIIIIIIIIIIIIIIIIIIIII
::: 7ra' and label the 2ischer "ro<ections for as"aragines.
a: Ho' many chiral centers are thereN IIIIIIIIIIIIIIIIIIIIIIIIIIII
@?
b: Ho' many total stereoisomers are thereN IIIIIIIIIIIIIIIIIIIIIIIII
c: ;hat 'ould be general formula for determining the ma$imum number of
stereoisomers 'hen n is the number of chiral center
:' 7ra' and label the 2ischer "ro<ections for tartaric acid. Mar* all the
chiral 'ith an ^. Circle the 2ischer "ro<ection and dra' its mirror "lane
for the meso com"ound.
a: Ho' many chiral centers are thereN IIIIIIIIIIIIIIIIIIIIIIIIIIII
b: Ho' many total stereoisomers are there IIIIIIIIIIIIIIIIIIIIIIIII
c: Ho' many "airs of enantiomers are thereN IIIIIIIIIIIIIIIIIIIIIIIIIIII
d: Ho' many "airs diastereomers are thereN IIIIIIIIIIIIIIIIIIIIIIIIIIIIIII
' 7ra' the 2ischer "ro<ection for your 23 33 ?3 @3 A% "entahydro$yhe$anal
and label all chiral centers 'ith an ^.
@@
a: Ho' many chiral centers are thereN IIIIIIIIIIIIIIIIIIIIIIIIIII
b: Ho' many total stereoisomers are "ossible based on general formula for isomersN
IIIIIIIIIIIII
c: Sho' your 2ischer "ro<ection to your instructor3 'hich 7%aldohe$ose did you
constructN
d: is the structure is straight or coiledN IIIIIIIIIIIIIIIIIIIIIIIIIII
e: ;hat class of com"ounds 'ould be made if the hydro$yl grou" on carbon @ reacts
'ith the
aldehyde grou"N
IIIIIIIIIIIIIIIIIIIIIIIIIIIIIIII
f: Ho' many atoms are in the ring of the resulting cyclic structureN
IIIIIIIIIIIIIIIIIIIIIIII
@A
@B
Carbohydrate
s
Learn the Dualitati-e tests to identify organic functional grou"s in carbohydrates
Learn the use of s"eci8c Dualitati-e tests to distinguish bet'een aldehydes and
*etones
Learn the Dualitati-e test for com"le$ carbohydrate
:ntroduction
Carbohydrates 'hich com"rise one of the three basic classes of foodstu>s3
contain carbon3 hydrogen and o$ygen atoms. ,hey are an im"ortant class of biological
macromolecules3 'hich are found in nature both in isolation as mono,) di,) and
polysaccharides and in association 'ith other biological macromolecules3 e.g.3 as
glycolipids and glycoproteins. ,hey are in-ol-ed in a di-erse range of "hysiological
roles3 such as molecular recognition3 energy storage en)yme regulation. Carbohydrates in
our diet are our ma<or source of energy.
(,glucose (,fructose Sucrose 6table sugar7
Carbohydrates are classi8ed as polyhydro&y aldehydes or polyhydro&y
ketones. ,herefore3 they 'ill e$hibit chemical "ro"erties associated 'ith both alcohols
and carbonyl com"ounds. Some e$am"les of carbohydrates are sho'n abo-e.
Carbohydrates easily cycli)e to form hemiacetals and hemi*etals. ndeed3 they s"end most
of their time in a cyclic form. ;hen cycli)ed3 the carbon that 'as the carbonyl carbon
becomes "art of the ring and is called the anomeric carbon. 6ou can easily 8nd the
anomeric carbon on any cyclic saccharide by locating the !=L6 carbon 'ith t'o o$ygen
atoms directly attached. Sucrose has t'o anomeric carbons3 as indicated 'ith asteris*s in
@C
M
M
the diagram abo-e.
f at least one of the anomeric carbons has a hydro$yl grou" directly attached3 it can
re-erse the cycli)ation "rocess and form the linear aldehyde or *etone again. n the linear
aldehyde or *etone form3 the molecule can "artici"ate in any aldehyde or *etone reaction.
<oes sucrose have a hydro!yl group directly attached to either anomeric carbon1 ;e are
going to do series of analyses to e$amine the reacti-ity of some monosaccharides3
disaccharides and "olysaccharides. ,ests similar to these may be used clinically to test for
glucose in urine and blood.
n the BenedictGs test a reducing sugar Ka sugar 'ith a hydro$yl directly
attached to an anomeric carbon: reacts 'ith the blue%colored Cu
2U
ion in the
"resence of base. ,he co""er K: ion is reduced to form co""er K: in a red%orange
Cu2! "reci"itate 'hereas the aldehyde grou" is o$idi)ed to the carbo$ylic acid
functional grou". n addition to all aldose monosaccharides gi-ing a "ositi-e
&enedictTs test3 *etose monosaccharides3 though lac*ing an aldehyde grou"3 react
due to the "resence of a hydro$yl grou" ne$t to the *etone grou". ,hus hydro$yl
*etones gi-e "ositi-e tests.

,he BarfoedGs Test is a -ariation of the redo$ reaction mentioned "re-iously.
Co""erK: acetate in acetic acid is not as reacti-e as the Cu
2U
&enedict s reagent.
,hus3 one can distinguish monosaccharides from disaccharides based on ho' fast
the red%orange "reci"itate forms. ,y"ically3 monosaccharides react 'ithin 2%3
minutes3 'hereas disaccharides ta*e longer.
,he SeliwanoKGs Test is used to distinguish from aldoses using the aromatic
alcohol in the "resence of concentrated hydrochloric acid. ,his is useful for both
monosaccharide *etose as 'ell as disaccharides *etose. # "ositi-e test is noted by
a red colored solution5 a yello' stra' or a"ricot color is not indicati-e of "ositi-e
test.
,o distinguish "entoses from he$oses one can use the BialGs Test. Pentoses react
'ith orcinol in the "resence of 2eCl3 and conc. HCl to gi-e a characteristic blue%
green color. =on%reacting sugars may "roduce a bro'n "reci"itate but the solution
usually remains the yello' color of the 2eCl3.
Starch is a com"le$ carbohydrate that interacts in the :odine Test. t is com"osed
of t'o fractions5 the linear3 helical fraction n and the branched amylo"ectine
fraction. ;hen 2 inserted into the interior of the amylase fraction3 a dar* blue color
is obser-ed.
E&perimental
6ou 'ill 8nd the follo'ing carbohydrate test solutions on your bench+ glucose3 galactose3
fructose3 arabinose3 maltose3 lactose3 sucrose and starch. These tests are ery colorful
and therefore your notebook should be full of obserations!
&enedictTs ,est+
1. Pre"are a boiling 'ater bath and label eight clean small test tubes.
2. n se"arate test tubes add 1 mL of the &enedictTs reagent. ,o each test tube add @
dro"s of the test carbohydrate solution. Mi$ the sam"les.
@E
3. Place all the test tubes at the same time into the boiling 'ater bath.
?. =ote and record the ho' long it ta*es for the red Cu2! "reci"itate to form5 also note if
the blue &enedictTs reagent color disa""ears.
@. #fter 10 minutes remo-e all the tubes and *ee" the boiling 'ater bath going for the
remaining three e$"eriments. 7id any sugars not "roduce the red "reci"itateN ;hich
are reducing sugarN ;hich are notN
&arfoedTs ,est+
1. .se the boiling 'ater bath from before and label a ne' set of C clean small test tubes.
2. n se"arate test tubes add 1 mL of the &arfoedTs reagent. ,o each test tube add 10
dro"s of the test carbohydrate solution. Mi$ the sam"les.
3. Place all of the test tubes at the same time into the boiling 'ater bath.
?. =ote and record the long it ta*es for the red Cu2! "reci"itate to form.
@. #fter 10 minutes remo-e all the tubes. 7etermine 'hich are monosaccharides3 'hich
are disaccharides
Seli'ano>Ts ,est+
1. .se the boiling 'ater bath from before and label a ne' set of C clean small test tubes.
2. n se"arate test tubes add Seli'ano>TTs reagent. ,o each test tube add 3 dro"s of the
test carbohydrate solution. Mi$ the sam"les
3. Place all of the test tubes at the same time into the boiling 'ater bath. =ote and record
ho' long it ta*es for the 8rst clear red colored solution to form.
?. Remo-e all of the test tubes as soon as the 8rst "ositi-e test is seen as "rolong heating
Kin e$cess of @ minutes: may cause s"urious results. ;hich sugar solutionKs: contain a
*etoseN
&ialTs ,est+
1. .se the boiling 'ater bath from before and label ane' set of C clean test tubes.
2. n se"arate test tubes add 1 mL of the &ialTs reagent. ,o each test tubes add 10 dro"s
of the carbohydrate solution.
3. Place all the test tubes at the same time into the boiling 'ater bath.
?. =ote and record ho' long it ta*es for the 8rst clear blue%green solution to form
@. Remo-e all the test tubes as soon the 8rst "ositi-e test is seen as "rolonged heating
may cause s"urious results. ;hich sugar solutionKs: contain "entoseN
odine test+
1. Place 3 dro"s of each test carbohydrate solution in se"arate 'ells of a clean s"ot "late.
A0
2. #dd 1 dro" of the iodine solution to each test carbohydrate solution. =ote and record
the color of each sam"le.
3. 7id any other solutions besides the starch solution gi-e a "ositi-e testN
A1
RE02RT SHEET,Carbohydrates
1. Com"lete the table of results. &e sure to include color_+
o
d
i
n
e

,
e
s
t
&
i
a
l
T
s

,
e
s
t
S
e
l
i
'
a
n
o
>
T
s

,
e
s
t
&
a
r
f
o
e
d
T
s

,
e
s
t
&
e
n
e
d
i
c
t
T
s

,
e
s
t
A2
C
a
r
b
o
h
y
d
r
a
t
e
G
l
u
c
o
s
e
G
a
l
a
c
t
o
s
e
2
r
u
c
t
o
s
e
#
r
a
b
i
n
o
s
e
M
a
l
t
o
s
e
L
a
c
t
o
s
e
S
u
c
r
o
s
e
S
t
a
r
c
h
2. Su""ose you sa' no sign of a color change in &enedictTs test3 no sign of a red
"reci"itate after 10 minutes 'ith &arfoedTs test3 and a dar* red colored solution
'ith the Seli'ano>Ts test. ;hat sugarKs: could you ha-eN
3. Su""ose you sa' a red "reci"itate 'ith the &enedictTs test3 a red "reci"itate after
2 minutes 'ith the &arfoedTs test3 and stra'%colored solution after more than @
minutes 'ith the Seli'ano>Ts test. ;hat sugarKs: could you ha-eN
?. Su""ose you sa' a red "reci"itate 'ith &enedictTs test3 no sign of red "reci"itate
after 10 minutes 'ith &arfoedTs test3 and a stra'%colored solution after more than
@ minutes 'ith Seli'ano>Ts test. ;hat sugarKs: could you ha-eN
A3
Acid,Base
Reactions with
Carbo&ylic Acids
and Esters
Learn about t'o -ery im"ortant functional grou"s3 carbo$ylic acids and esters
Learn about the hydrolysis reaction of ester3 and itTs "roduct
Learn about the industrial a""lication of sa"oni8cation
:ntroduction
Salicylic acid Methyl ben)oate
n the 8rst "art of this e$"eriment you are going to e-aluate the solubility of salicylic
acid in cool 'ater and hot 'ater. ,hen3 the reacti-ity of salicylic acid 'ith aDueous =a!H
'ill be in-estigated. #s you 'ould e$"ect carbo$ylic acids should react 'ith 'ater to form a
'ater%soluble carbo$ylate anion. n general the follo'ing acid%base reaction occurs+
R%C!!H U H
2
! R%C!!
%

KaD:
U H
3
!
U
'ae care) solubility is also dependent of the si:e of the alyl or aryl group attached to the
carbo!ylic acid functional group. #dditionally3 carbo$ylic acids react 'ith bases to form
'ater soluble salts as sho'n belo'+
RC!!H U =a!H
KaD:
RC!!
%
KaD:
U =a
U
KaD:
U H
2
!
"ote) salicylic acid contains, in addition to the carbo!ylic acid functional group, a phenol
group hydrogen that can also react with the base.
,he resultant sodium carbo$ylateF"henolate salt can further react 'ith strong acids to
reform the free acid and "henol as sho'n belo'+
A?
R%C!!
%
U HCl
KaD:
R%C!!H
Ph%C!!
%
U HCl
KaD:
Ph%C!!H
,he use of bases and acids ser-es as Gsolubility s'itchesH to con-ert an insoluble
form of a com"ound to a soluble form and -ice -ersa. !bser-ing the solubility or
insolubility of reactants and For "roducts ser-es as a means to monitor acid base reactions.
Saponifcation is the lysis of an ester 'ith a strong base to form an alcohol and the
salt of a carbo$ylic acid. Sa"oni8cation is commonly used to refer to the reaction of a
metallic al*ali Kbase: 'ith a fat or oil to form soa". ,he conce"t of a solubility s'itch 'ill
also ser-e as the basis for follo'ing the base K=a!H: cataly)ed hydrolysis Ks"eci8cation:
reaction of an ester Kmethyl ben)oate: to the subseDuent 'ater soluble carbo$ylate salt
Ksodium ben)oate: and alcohol Kmethanol:. ,he resultant mi$ture containing the sodium
ben)oate is then reacted 'ith acid to form ben)oic acid.
E&perimental
Solubility and #cid%&ase Reactions of Salicylic #cid+
1. Set u" a boiling 'ater bath 'ith a 2@0mL bea*er on a hot "late and an ice 'ater
bath in a bea*er.

2. Place a small amount of salicylic acid K"ea si)e amount: into a clean small test tube
and add a @mL of distilled 'ater. Stir 'ell and record your obser-ations in your
noteboo*.
3. Carefully "lace the test tube into the boiling 'ater bath. Record your obser-ations.

?. Remo-e the test tube and cool the solution in the ice 'ater bath. =ote that you may
ha-e to scratch the inside o surface of the test tube 'ith a glass rod. 6our instructor
'ill demonstrate this techniDue. Record your obser-ations.
@. #dd 3M =a!H to the mi$ture dro" by dro" and agitate by 8nger 4ic*ing three test
tubes after each dro". &e sure to *ee" trac* of the number of dro"s that you add.
Record your obser-ations and 'rite the o-erall eDuation for this reaction.
A. 2inally3 add to the solution as many dro"s of 3M HCl as you used for the 3M =a!H.
,hen add se-eral more dro"s of the 3M HCl. Record your obser-ations and 'rite the
o-erall eDuation for this reaction.
#cid%&ase Reactions of Methyl &en)oate+
1. Place 3 dro"s of methyl ben)oate into a clean test tube and add 2 mL of distilled
'ater. Record your obser-ations in your noteboo*.

2. #dd 12 dro"s of 2.@ M or 3M =a!H and mi$ the contents of the test tube. Place the
test tube into the boiling 'ater bath for 30 minutes Kor longer: and e-ery @ minutes
stir the contents -igorously 'ith a clean glass rod3 re"lacing the test tube bac* into
the boiling 'ater bath. Sto" the reaction 'hen you <udge the solution to be
homogeneous. Record your obser-ations and 'rite the o-erall eDuation for this
reaction.

A@
3. Cool the mi$ture to room tem"erature by running cold ta" 'ater along the outside
of the test tube.
?. #dd 1@ dro"s of 3M HCl mi$ing by 8nger 4ic*ing the test tube after each addition.
Record your obser-ations and 'rite the o-erall eDuation for this reaction.
AA
RE02RT SHEET,Acid,Base R&ns with Carbo&ylic Acids and
Esters
Solubility and #cid%&ase Reactions of Salicylic #cid+
1. 7oes salicylic acid dissol-e in the cold 'aterN

2. 7oes salicylic acid dissol-e in hot 'aterN
3. 7oes salicylic acid dissol-e in aDueous sodium hydro$ideN
?. ;rite an o-erall eDuation for the reaction of salicylic acid 'ith aDueous sodium
hydro$ide.
@. ;hat did you obser-e 'hen hydrochloric acid 'as added to the test tube during ste" AN
A. ;rite an o-erall eDuation for the chemical reaction that occurs in ste" A.
#cid%&ase Reactions of Methyl &en)oate+
1. ;rite an o-erall chemical eDuation for the sa"oni8cation of methyl ben)oate.
AB
2. #re the initial "roducts of the sa"onication of methyl ben)oate soluble or insoluble in
'aterN
3. ;hat did you obser-e 'hen hydrochloric acid in ste" ?N
?. ;rite an o-erall chemical eDuation for the reaction for ste" ?.
AC
Synthesis of
Aspirin
Learn to do the organic synthesis of an ester from an alcohol and an anhydride
Learn ho' to "urify the "roduct in the organic synthesis
Learn ho' to calculate the Z yield of a "roduct
Learn di>erent techniDues to com"are the "urity of a synthesi)ed "roduct 'ith
commercial "roduct.
:ntroduction
#cetylsalicylic acid K#S#:3 commonly called as"irin3 is 'idely used as medicine to
reduce fe-er Kan anti"yretic:3 to reduce "ain Kan analgesic:3 to reduce s'elling Kanti%
in4ammatory:3 and to "re-ent "latelet aggregation that initiali)es thrombosis or hemostasis
Kanti%clotting:. #s"irin inhibits the en)ymes necessary for the formation of "rostaglandins
and thrombo$anes Khormones: that are associated 'ith "ain3 fe-er3 in4ammation3 and
blood%clotting in the human body. t has been also suggested that as"irin small amount as3
C0%100mg for daily ingestion can lo'er the ris* of stro*e and heart attac* in at%ris* adults.
#s"irin is an ester of acetic acid and salicylic acid. Salicylic acid3 is acting as the
alcohol3 because this also has a hydro$yl grou" attached to the ben)ene ring besides a
carbo$ylic acid. ,hough esters can be "roduced from the direct esteri8cation of an alcohol
and a carbo$ylic acid in the "resence of an acid catalyst3 ty"ically sulfuric acid3 the "resent
method to "re"are as"irin uses acetic anhydride a deri-ati-e of acetic acid to form more
Duic*ly an acetate ester 'ith salicylic acid. #cetic anhydride3 as a substitute acetylating
agent reacts 'ith salicylic acid as follo's+
n the 8rst 'ee* of a t'o%'ee* e$"eriment you 'ill synthesis as"irin Kacetyl salicylic
acid:. ,he follo'ing 'ee* you 'ill determine the "ercent yield of your synthesi)ed as"irin.
AE
,hen you 'ill analy)e the "urity of your "roduct by thin layer chromatogra"hy and by a
melting "oint determination.
CH#R#C,ER`#,!= !2 S6=,HES`E7 #CE,6LS#LC6LC #C7 K#S#3 as"irin:+
,he second 'ee* you 'ill determine the "ercent yield of the synthesi)ed "roduct3
as"irin and "erform di>erent techniDues to determine the "urity of the "roduct. !f course3
this as"irin is not suitable for oral admission or any "hysiological test because reagents
used for this synthesis are not of suJcient "urity for ingestion.
7etermination of Percent 6ield+
Percent yield is de8ned as follo's+
Z yield S mass of acetylsalicylic acidKactual yield of #S# in e$"eriment:
$ 100
,heoretical yield of #S# Kcalculated from stoichiometric
relationshi":
,he actual yield is the number of grams of acetyl salicylic acid that you actually
made in the laboratory. ,he theoretical yield is the number of grams of acetylsalicylic acid
you should get based on the stoichiometry of the chemical eDuation. ,he theoretical yield
is based on the number of grams of salicylic acid you used. 2irst con-ert the number grams
of salicylic acid to moles by di-iding the grams of salicylic acid by the molecular 'eight of
salicylic acid K13C gFmol:. !ne mole of acetylsalicylic acid is formed for each mole of
salicylic acid used. ,herefore3 the number of moles of acetylsalicylic acid is the same as
the number of moles of salicylic acid. ,o con-ert to grams of acetyl salicylic acid3 multi"ly
the number of moles you calculated by the molecular 'eight of acetylsalicylic acid K1C0
gFmol:
,hin Layer Chromatogra"hy+
!ne 'ay of determining the "urity of your "roduct is to do thin layer
chromatogra"hy using your "roduct3 one of the reactant Ksalicylic acid: and an authentic
as"irin sam"le. ,his is the same "rocedure you com"leted 'hen you analy)ed chloro"hyll
and carotenoids from s"inach. Re-ie' that e$"eriment if you ha-e forgotten.
7etermination of the melting "oint+
6ou should use the Meltem" a""aratus to determine the melting "oint of your
acetylsalicylic acid. Carefully "lace a thermometer in the slot on the Meltem" de-ice.
,hese are mercury thermometers and are Duite fragile3 so handle the thermometer 'ith
care. f you brea* the thermometer3 notify the instructor immediately so the mercury s"ill
can be cleaned u". &efore measuring the melting "oint of your "roduct3 loo* u" the true
melting "oint of acetylsalicylic acid in ,he Handboo* of Chemistry and "hysics. ,his 'ill
gi-e you an idea of about 'here your "roduct should melt.
2erric chloride test+
7issol-e a small amount of your "roduct in 'ater and test it 'ith ferric chloride. #
"ur"le color indicates the "resence of a "henol Kunreacted salicylic acid: in your "roduct.
Com"are your results 'ith those obtained from a sam"le of authentic as"irin.
B0
#cid hydrolysis of acetylsalicylic acid+
Esters can be hydroly)ed by heating them in the "resence of an acid. ,he original
alcohol and carbo$ylic are generated from the acid hydrolysis.
E&perimental
Synthesis+
1. Place Y @0 mL of distilled 'ater into a 2@0 mL bea*er3 add a cou"le of boiling chi"s
and heat to boiling. #lso "ut Y20 mL of distilled 'ater in a @0 mL Erlenmeyer 4as*
into an ice 'ater bath using another 2@0 mL bea*er K2ig. #3 =ote+ use of hot "late
'ill be more safe than using burner:.
2. ;rite your grou" number in "encil on a "iece of 8lter "a"er. ,are the balance and
'eigh the 8lter "a"er to the nearest 0.001 g5 record this -alue in your noteboo*.
!btain Y 1 g of salicylic acid and "lace it into a tarred "lastic 'eigh boat. ;eigh the
salicylic acid to the nearest 0.001 g and record this -alue in your noteboo*.
3. Place the salicylic acid into a clean dry medium test tube. #dd 2 mL of acetic
anhydride
and 2 dro"s of concentrated sulfuric acid Kcaution+ acetic anhydride "roduces
irritation and necrosis of tissues in liDuid or in -a"or state:. #-oid contact 'ith s*in
and eyes. 7o this addition in the hood.
?. Place the test tube containing the reaction mi$ture into the boiling 'ater bath. Stir
the mi$ture -igorously 'ith a clean glass rod 'hile in the boiling 'ater bath. &e
careful not to brea* the test tube.
@. #fter the entire solid has dissol-ed3 remo-e the test tube into the ice 'ater bath. f
the crystals do not form3 induce crystalli)ation by scratching the inside of the test
tube 'ith a glass rod. ;hen crystalli)ation is com"lete add 10 mL of the ice cold
'ater.
A. 2old your 'eighed3 initialed 8lter "a"er so that it is 4uted Kyour instructor 'ill
demonstrate this: and "lace it into a short stem funnel.
B. Collect the solid on the 8lter "a"er in the funnel. Rinse the solid 'ith 2 or 3 small K@
mL: "ortions of ice cold 'ater. &e sure to let the 'ater drain through the 8lter
bet'een additions of the rinses.
C. Carefully remo-e the 8lter "a"er from the funnel and s"read it out on a "iece of
"a"er to'el. Set the "a"er to'el 'ith the 8lter "a"er in a dra'er and allo' it to dry
until the ne$t laboratory "eriod.
E. 7etermine the gram formula 'eight for salicylic acid. .sing the mass of your
salicylic acid3 determine the number of moles in the reaction. 7etermine the gram
formula 'eight for the acetyl salicylic acid "roduct. 2inally3 calculate the mass of
"roduct e$"ected if all of the salicylic acid is con-erted to acetyl salicylic acid.
10. 6our instructor 'ill demonstrate ho' to "re"are a sam"le for a melting "oint
a""aratus. Practice ta*ing melting "oints using the salicylic acid.
B1
11. ,he follo'ing 'ee* carefully remo-e the 8lter "a"er containing the as"irin from the
dra'er and 'eigh it to the nearest 0.001 g. Record this -alue in your noteboo*.
12. 7etermine the "ercent of the of yield of the "roduct as"irin. Sa-e as"irin for further
analysis.
,hin layer chromatogra"hy+
1. !btain Y 1 mL of methanol to three se"arate clean and dry test tubes. nto
these se"arate test tubes3 add a small amount of your synthesi)ed as"irin3
commercial as"irin and salicylic acid3 'hich you ha-e used for the synthesis. &e
sure label each test tube so that you *no' 'hich is contained in each test tube.
,o aid in dissol-ing the crush the solid gently 'ith a clean dry glass rod. &e sure
to clean the glass rod "rior to using for the ne$t sam"le.
2. !btain B cm a 1? cm 4uorescent silica gel ,LC "late for your grou".3 2ollo' the
"rocedure in "ages 2?3 2@3 and 2A to s"ot K? bL: your "roduct3 authentic as"irin
and salicylic acid. &e sure to mar* care fully the s"ot line at the bottom and
label for the s"ots at the to". &e sure to *ee" at least K10 mm: s"ace bet'een
the s"ots. 6our "roductTs s"ot should be on the left most "osition3 then the s"ot
of authentic as"irin on the middle "osition and salicylic acid s"ot on the right
most "osition. ,ry to *ee" the diameter of the s"ot less than 2 mm. Se-eral
Duic* small a""lications 'ill be better than one large a""lication.
3. Pre"are a ,LC de-elo"ing tan* similar to 'hat you used "re-iously. Pleace one
half of a 8lter "a"er into the <ar3 then add 10 mL of the ,LC sol-ent "ro-ided by
your instructor. Ma*e sure the 8lter "a"er is com"letely saturated 'ith the
de-elo"ing sol-ent. ;hen the three sam"les ha-e been a""lied3 carefully "lace
the ,LC "late into the de-elo"ing chamber. Ma*e sure that the sol-ent le-el is
belo' your original s"ots.

?. #fter the sol-ent front reaches 1 cm from the to" of the ,LC "late3 remo-e the
"late from the bea*er. #llo' it to dry under the hood. .se a ./ lam" to -ie' the
s"ots. Carefully3 mar* the s"ots 'ith a "encil. Com"are the s"ots 'ith one
another. 7ra' a re"roduction of your de-elo"ed chromatogram in your lab
noteboo*.
Melting "oint determination+
1. Place a small amount of your "roduct into a melting "oint ca"illary tube as
demonstrated by your instructor. Place the ca"illary tube into the ca"illary slot
on the Meltem" a""aratus.

2. ,o obtain an accurate melting "oint it is necessary to heat the sam"le slo'ly. 2or
the melting "oint of your "roduct3 the meltem" should be set at about ?0/. ,urn
on the "o'er and 'atch the crystals through the magnifying glass of the
Meltem" de-ice.
3. #s soon as the crystals start to melt3 record the tem"erature as "recisely as
"ossible5 Qee" 'atching the crystals. Record a second tem"erature as soon as
all of the crystals ha-e melted.
?. f your "roduct is "ure3 the melting range Kthe di>erence bet'een the t'o
recorded tem"eratures: 'ill be small%"erha"s a degree or t'o. f your "roduct is
not "ure3 you 'ill ha-e a 'ider melting "oint range. f your "roduct is "ure3 the
B2
melting "oint should be similar to the true -alue for acetylsalicylic acid. f it is
im"ure3 your 'ill ha-e melting "oint lo'er than the true -alue.
2erric chloride test+
7issol-e a small amount of your "roduct in Y 1 mL of 'ater. #dd t'o dro"s of ferric
chloride solution and note the color. Re"eat 'ith an authentic as"irin sam"le of
as"irin and 'ith the salicylic acid that you ha-e used. Record your obser-ations in
your lab noteboo*.
#cid hydrolysis of acetylsalicylic acid+
7issol-e a small amount of your "roduct in Y 1 mL of 'ater. #dd 8-e dro"s of conc.
Sulfuric acid and heat the test tube in a boiling 'ater bath for 10%1@ minutes.
Remo-e the test tube3 cool to room tem"erature and test for the "resence of a
"henol by adding t'o dro"s of ferric chloride.
B3
B?
RE02RT SHEET,Synthesis of Aspirin
Synthesis+
1. Mass of 8lter "a"erIIIIIIIIIIIIIIIIIIIIIIIIIIII
2. Mass of 8lter "a"er "lus "roductIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIII
3. Mass of acetylsalicylic acid Kas"irin:IIIIIIIIIIIIIIIIIIIIIIIIIIIIIIII
?. Mass of salicylic acid usedIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIII
@. Moles of salicylic acid usedIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIII
A. Moles of acetylsalicylic acid e$"ected Ksho' theoretical yield:Krefer to the eDuation:
B. Z yield of acetylsalicylic acid Ksho' calculation:
C. ;rite full chemical eDuation Kline%bond formulas for both reactants and "roducts: for
the synthesis of as"irin.
E. #fter 8ltering your reaction mi$ture3 'hat 'as the "ur"ose of rinsing the "roduct
'ith 'aterN
10. ;hy it is im"ortant for the 'ater to be ice coldN
B@
Characteri)ation+
1. ;rite the literature -alue for the melting "oint of acetylsalicylic
acid.IIIIIIIIIIIIIII
o
C
2. ;rite the melting "oint range of your "roductN IIIIIIIIIIII
o
C to IIIIIIIIIII
o
C
3. 7ra' a re"roduction of your de-elo"ed chromatogram. Sho' the "osition of all
s"ots3 the sol-ent front and s"otting line. ;hat conclusions can be dra'n about the
"urity of your "roduct from this chromatogramN
?. ;hat conclusion can be dra'n about the "urity of your "roduct from this
chromatogramN
@. ;hat did you obser-e 'hen 2eCl3 'as added to your "roductN
A. ;hat did you obser-e 'hen 2eCl3 'as added to authentic as"irinN
B. ;hat did you obser-e 'hen 2eCl3 'as added to salicylic acidN
C. ;rite the full chemical eDuation Kline%bond formulas for both reactants and
"roducts: for the hydrolysis of your acetylsalicylic acid.
E. ;hat did you obser-e 'hen 2eCl3 'as added to the as"irin hydrolysis solutionN
10. .sing all e$"erimental data3 com"are the "urity of your as"irin 'ith that of the
authentic as"irin sam"le.
BA

Synthesis @
properties of Soap
Learn about the "rocess of soa" ma*ing
Learn the "rocedure for "urifying and testing the of "ro"erties of a soa"
:ntroduction
# natural soa" is the sodium or "otassium salt of long chain fatty acids "roduced
by the base cataly)ed hydrolysis of triacylglycerol Kthe fat storage molecule in "lants and
animals3 *no'n clinically as GtriglyceridesH:. n the 8rst "art of this e$"eriment you 'ill
"re"are soa" by a sa"oni8cation reaction of a small sam"le of oil or fat. # generali)ed
sa"oni8cation reaction is sho'n belo'+
n the second "art of this e$"eriment some of the "ro"erties K"H and solubility: in
solutions of your soa" 'ill be e$amined.
E&perimental
Synthesis+
1. Caution must be obser-ed as the concentrated sodium hydro$ide Klye: is corrosi-e
and can cause burns to s*in3 destruction of clothing and irre-ersible cornea damage
to the eye. #t no time are your safety glassesFgoggles to be remo-ed during this
e$"eriment.
2. Pre"are boiling 'ater bath in a A00 mL bea*er5 be sure to add a cou"le of boiling
chi"s. Place 12 mL of oil or 10 g of fat into a 2@0 Erlenmeyer 4as*. #lso "re"are an
ice 'ater bath using another A00 mL bea*er.
3. #dd 10 mL of ethanol Kethylalcohol: and 12 ml of AM sodium hydro$ide to the
-egetable oil. ^^6ou may add a small "iece of 'a$ crayon no'3 if you 'ant your
soa" to ha-e a color. Many dyes used in crayons 'ill be altered due to the change
BB
in "H. ,he "ro"er color should return 'hen you rinse your soa" later. Someone in
your grou" needs to lea-e theirs 'hite for the tests

?. Stir the mi$ture 'ith a glass stirring rod. .sing a ring stand and a clam" secure the
4as* and heat it in the boiling 'ater bath. Continuously stir the mi$ture during the
heating "rocess to "re-enting the mi$ture from foaming u" the sides of the 4as*.
@. Heat the mi$ture in the 'ater bath3 'ith stirring until the odor of ethyl alcohol is no
longer detected. ,his may ta*e 1@ to 30 minutes. Remo-e the 4as* from bath.
A. Place the 4as* into the ice bath. Cool the soa" solution for 10 minutes.
B. ,o the contents of the 4as* add 20 mL of a concentrated sodium chloride solution.
.sing a s"atula3 brea* u" the lum"s of soa" as com"letely as "ossible to "ermit
contact bet'een the solid and the sodium chloride 'ash solution. Carefully decant
the solution to remo-e the 'ash solution 'hile retaining the solid in the 4as*.
C. Re"eat the 'ashing and decanting ste" t'o more times. #fter the 'ashing remo-e
the last traces of liDuid by dum"ing the solid into the "a"er to'els and carefully
blotting the soa" 'ith additional "a"er to'els. #-oid touching the soa" 'ith your
s*in.
#nalysis+
1. 7issol-e a small "ea si)ed of your soa" in a small test tube containing @ mL of distilled
'ater. #dd 3 to ? dro"s of .ni-ersal indicator. =ote the color a""eared. .sing the
.ni-ersal indicator reference card3 determine the a""ro$imate "H of your soa"
solution. Re"eat this e$"eriment using a "uri8ed commercial soa" Ke.g.3 -ory:. Record
color and "H -alue in your noteboo*.
2. n three se"arate clean test tubes "lace @ mL each of distilled 'ater3 ta" 'ater and
10Z CaCl2 Kcalcium chloride : . #dd a small "ea si)ed "ieces of your soa" to each
se"arate test solution. Ma*e sure that you use eDual si)ed amounts. Sto""er each tube
and sha*e them -igorously. 7escribe the relati-e amounts of lather and foam that
a""ear in each tube. Record your obser-ations in your noteboo*.
3. 7issol-e a small amount of your soa" in a minimum amount of distilled 'ater5 estimate
the -olume of 'ater you used. ,o the dissol-ed soa" solution add an eDual -olume of
concentrated sodium chloride solution. 7escribe and record in your note boo* 'hat
ha""ens.
?. /isit other grou"s and com"are the te$ture of soa"s made from di>erent fat sources.
BC
RE02RT SHEET,Synthesis and 0roperties of Soap
1. ;rite a chemical eDuation for the sa"oni8cation of a triglycerol that contains
"almitic acid3 oleic acid and linoleic acid as the three fatty acid moieties.
2. ;hat 'as the "ur"ose of adding the concentrated salt solution to your soa"
"re"arationN
3. a:. ;hat 'as the "H of your soa" solutionN
b:. ;as your soa" solution acidic3 basic or neutralN
c: ;hat 'as the "H of the commercial soa" solutionN
d: ;as the commercial soa" solution acidic3 basic or neutralN
e: &ased on your result to "arts 3a%3d abo-e is it "ossible to ha-e a neutral solution
of a "ure soa" in distilled 'aterN
?. 7escribed the obser-ed beha-ior 'hen your soa" 'as added to each of the
follo'ing and sha*en u"+
a: 7istilled 'ater+
b: ,a" 'ater+
BE
c: Calcium chloride solution+
@. ;rite a net eDuation for the reaction of calcium ions 'ith the anion of "almitic acid.
A. ;hat did you obser-ed 'hen concentrated sodium chloride solution 'as added to
dissol-ed soa"N
B. ;hat does the obser-ation in A3 suggest about the e>ecti-eness of ordinary soa" in
sea'aterN
C. ;ere there any di>erences in te$ture in soa" from di>erent fat sourcesN 7oes this
agree 'ith your *no'ledge about saturated -ersus unsaturated fatsN
C0
:solation and
Characteri%ation
of Casein from
?ilk
#do"ted from+ GIsolation of $rotein, +arbohydrate and =at from ;ilH3 Mohr. S. C.3 GriJn3 S.
2.3 and Gensler3 ;.c.3 in &aboratory ;anual for =undamentals of %rganic and >iological
+hemistry cohn McMurry and Mary E. Castellion3 Engle'ood Cli>s3 Prentice%Hall3 1EE?
;ayne P. #nderson K?F2002:
Learn the about the "rotein "resent in mil*3 and cheese
Learn the "rocedure to isolate the "rotein from the mil*
Learn the techniDues used to characteri)e the "rotein
:ntroduction
6ou may recall the Mother Goose nursery rhyme3 GLittle Miss Mu>et sat on a tu>et3
eating her curds and 'heyX..H ;hen mil* is acidi8ed3 it is transformed into a solid
com"onent3 called curd3 and a liDuid com"onent called 'hey. ,his method is used to
ma*e cottage cheese. ,he curds contain butterfat and a "rotein called casein3 'hich
contain all of the common amino acids and is "articularly rich in the essential ones.
Casein e$ist in the mil* as a soluble calcium salt3 that "reci"itates at "H -alues belo'
?.A. So mil* can be curdled by acids such as lactic acid that forms during natural
souring of mil*. ,he carbohydrate3 lactose3 is "resent in the 'hey. n this e$"eriment
you 'ill also isolate casein from mil* and carry out some Dualitati-e tests for "rotein.
,he &iuret test is generally used for "rotein. ;hen the "ale blue Cu
2U
ion forms a
com"le$ 'ith ad<acent amide nitrogen of the "e"tide bac*bone3 a -ery dee" -iolet blue
color results.
C1
,he Ranto"eroteic acid test is on the other is a general test for the "resence of the
aromatic amino acids3 try"to"han3 "henylalanine and tyrosine3 in "roteins. #romatic
grou"s that ha-e an amino grou" Ktry"to"han: or a hydro$yl grou" Ktyrosine: are easily
nitrated by concentrated nitric acid to form yello' K$antho3 Gree* for yello': colored
aromatic nitro com"ounds.
E&perimental
1. 7etermine the mass of a 12@ mL Erlenmeyer 4as*. #dd @0 mL of mil* to the 4as*
and re%'eigh the 4as* to determine the mass of the mil*. Chec* the label on the
mil* container and record the amount of "rotein "er ser-ing in your noteboo*.

2. Pre"are a 'ater bath by "lacing 200mL of 'ater in a A00 mL bea*er. Heat the 'ater
bath to ?0
o
C5 as the tem"erature is critical for this e$"eriment monitor the
tem"erature 'ith a thermometer. Place the 4as* containing the mil* into the 'ater
bath.
3. Slo'ly add 10 dro"s of glacial acetic acid to the mil* 'hile stirring 'ith a glass rod.
Continue to add dro"s of glacial acetic acid dro" 'ise until no more "reci"itate is
formed 'hen a dro" of acid is added. #llo' the mi$ture to cool.
?. 2ilter the mi$ture into a 2@0 mL bea*er by "ouring it through cheese cloth that has
been fastened to the bea*er 'ith a rubber band. SDuee)e out as much liDuid as
"ossible from the solid. ,hen scra"e the solid into a 100 mL bea*er.
@. ,o remo-e any fat from the curd Kdo you e$"ect any for s*im mil*N:3 add 2@ mL of
ethanol to the solid in the 100 mL bea*er. Stir the mi$ture for about @ minutes5 then
let the solid settle. ,he fat 'ill dissol-e in the alcohol. 7ecant the liDuid into another
bea*er.
A. .nder a hood3 add 2@ ml of a 1+1 K-F-: mi$ture of diethyl ether and ethanol to
residue. &e sure that that there is no 4ames or s"ar*s "resent as the diethyl ether is
e$tremely 4ammable. Stir the mi$ture for about @ minutes. Let the "rotein solid dry
in your dra'er for a 'ee*3 then 'eigh your solid and determine the Z yield during
the ne$t class "eriod.
&.RE, ,ES,+
1. #dd a "ea si)ed amount of your casein to a small test tube and dissol-e it in ? mL of
distilled 'ater. 7i-ide this "rotein solution into t'o 2 mL "ortions. Sa-e one "ortion
for the ne$t test.
2. ,horoughly mi$ 2mL of the "rotein solution 'ith 2 mL of 3M sodium hydro$ide
solution. #dd 1 dro" of 1Z of co""er sulfate solution. =ote the color and record
your obser-ations in your noteboo*.
3. Continue to add the co""er sulfate solution one dro" at a time3 note and record your
obser-ations. Sto" after adding 10 dro"s of the co""er sulfate solution.
?. Re"eat this test 'ith a 1Z casein solution.
R#=,H!PR!,EC ,ES,+
C2
1. ,o the second 2 mL "rotein solution carefully add 1 mL of concentrated nitric acid.
Mi$ and note the a""earance of any hea-y 'hite "reci"itate.
2. ;arm the mi$ture carefully in a hot 'ater bath noting any change to a yello'
colored solution.
3. Cool the mi$ture in a stream of cold ta" 'ater and carefully add a fe' dro"s of 3M
sodium hydro$ide. # "ositi-e test is indicated by the yello' color changing into
orange color. ,he entire tube does not ha-e to turn to orange. Loo* for the color as
the sodium hydro$ide is added to the solution or on a "iece of "reci"itate on the
'all of the test tube.
?. Re"eat this test 'ith 1Z casein solution.
C3
C?
RE02RT SHEET,:solation and Characteri%ation of Casein
from ?ilk
solation+
1. Mass of 12@ mL Erlenmeyer 4as* IIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIII
2. Mass of 12@ mL Erlenmeyer 4as* Umil* IIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIII
3. Mass of mil* IIIIIIIIIIIIIIIIIIIIIIIIIIIIII
?. Mass of crude casein Kmay not be totally dry: IIIIIIIIIIIIIIIIIIIIIIIIIIIIIII
@. Calculate Z yield Ksho' your calculation:+
A. 7etermine the amount of "rotein in a ser-ing of mil* Kuse the mass of crude casein:
Ksho' your calculation:
&iuret test+
B. Color of 0.1Z of co""er sulfate solution+
C. Color of your casein U sodium hydro$ide solution+
E. Color of your casein solution after addition of one dro" of co""er sulfate+
10. Color changes obser-ed after adding additional dro"s of co""er sulfate+
11. Color of 1Z casein U sodium hydro$ide solution+
C@
12. Color of your 1Z "ro-ided casein solution after addition of one dro" of co""er
sulfate+
13. Color changes obser-ed after adding additional dro"s of co""er sulfate+
Rantho"roteic acid test+
1?. !bser-ation for your casein mi$ed 'ith concentrated nitric acid+
1@. !bser-ation for your casein mi$ed 'ith concentrated nitric acid after heating+
1A. !bser-ation for your casein after adding 3M sodium hydro$ide+
1B. !bser-ation for 1Z casein mi$ed 'ith concentrated nitric acid+
1C. !bser-ation for 1Z casein 'ith concentrated nitric acid after heating+
1E. !bser-ation for 1Z casein after adding 3M sodium hydro$ide+
Mil* com"arison+
7id one of the -arieties of mil* ha-e a higher "ercentage by 'eight of "roteinN
CA
Amylase" The
Acti*ity of an
En%yme
#da"ted from+ G2actorTs #>ecting En)ymatic #cti-ityH in cohn R. Holum and Sandra L.
!lmstead3 &aboratorty manual for =undamentals of ?eneral, %rganic and >iological
+hemistry3 @
th
Ed.3 =e' 6or*+ ;iley3 1EE?. Michael E. Pugh and ;ayne P. #nderson KRe-.
3F200?:
Learn about en)ymes that cataly)e biological reactions
Learn about the factors that in4uence the acti-ity of en)ymes
:ntroduction
#mylase3 an en)yme that is found in sali-a3 cataly)es the hydrolysis of starch
Kamylase:. Since en)ymes are "roteins3 their secondary and tertiary structures are
e>ected by tem"erature3 "H3 and the "resence of hea-y metal ions. En)yme acti-ity is
closely associated 'ith the structure of an en)yme. So any change in the secondary or
tertiary structure leads to a change in en)ymatic acti-ity.
n this e$"eriment you 'ill e$amine the e>ect of tem"erature and "H on the acti-ity
of amylase. Molecular iodine forms a com"le$ 'ith starch that has a characteristic dee"
blue color. #s starch undergoes hydrolysis to form oligosaccharides and glucose3 the
characteristics color of the starch Piodine com"le$ disa""ears. ,herefore3 loss of the
dee" blue color can be used to measure of the e$tent of hydrolysis of starch. # second
test for hydrolysis is the occurrence of a "ositi-e &enedictTs test for the solution. Starch
is not a reducing sugar3 but glucose formed u"on hydrolysis is a reducing sugar.
E&perimental
P#R, . E>ect of ,em"erature on En)yme #cti-ity
n order to ma*e sure that the concentration s of en)yme and starch remain
reasonably constant in di>erent "arts of the e$"eriment3 use an eye dro""er to measure
Duantities of solutions. #ssume that 20 dro"s re"resent 1.0 mL.
CB
1. Label three medium test tubes as G0H3 GrtH and G100H. nto each test tube "lace bu>ered
solution and 2.@ mL of distilled 'ater. Place the test tube mar*ed G0H into the ice bath3
the one mar*ed GrtH into room tem"erature and G100H into a bea*er of boiling 'ater.
2. Place 1 mL of freshly "re"ared amylase solution K100 mgF100 mL: into each of three
small test tubes. Put one of these into the ice bath3 another into the thermostated
'ater bath3 and the third into the bea*er of boiling 'ater.
3. #llo' the solutions to remain in the tem"erature baths for about @%10 minutes to
eDuilibrate.
+arry out the following steps in se(uence for the solutions in the 0
o
, rt, and @00
o

temperature baths.
?. Remo-e the test tube containing the starch and the test tube containing the en)yme
from the bath and "our the en)yme solution into the starch solution. Record the time to
the nearest second3 and call this starting time 0.

@. Luic*ly "lace a "iece of "ara8lm o-er the end of the test tube3 mi$ thoroughly3 remo-e
the "ara8lm and remo-e a small amount of the solution from the test tube using a
dis"osable "i"ette. Place the remaining solution bac* into the tem"erature bath. Place
? dro"s of the solution onto a s"ot "late that contain 1 dro" of the iodine solution3
record the color. Put any e$cess starchFen)yme solution in the "i"ette bac* into the
test tube in the tem"erature bath.
A. ,a*e a sam"le from the solution using the dis"osable "i"ette e$actly 1 minute follo'ing
time 0. Place ? dro"s onto a second s"ot of the s"ot "late containing 1 dro" of iodine
solution3 record the color. Return any e$cess starchFen)yme solution in the "i"ette to
test tube in the tem"erature bath.
B. Re"eat the "rocedure in at the follo'ing time inter-als until the color of the solution
follo'ing addition of the iodine is yello'+ 2 min3 ? min3 A min3 C min3 and 10 min. f the
color of the solution in the s"ot "late remains yello' for successi-e trials. t is not
necessary to continue the run.
Part . E>ect of "H on En)yme #cti-ity
1. Label three medium test tubes as G@H3 GBH and GEH.
2. nto the test tube mar*ed G@H3 "lace 2.@ mL of unbu>ered starch solution and 2.@
mL of "H @ bu>er solution. nto the test tube mar*ed GBH "lace 2.@ mL of unbu>ered
starch solution and 2.@ mL of "H B bu>er solution. nto the test tube mar*ed GEH
"lace 2.@ mL of unbu>ered starch solution and 2.@ mL of "H E bu>er solution. Place
these into either the room tem"erature bath or the ice bath3 de"ending on 'hich
tem"erature ga-e the better results in "art .
3. Place 1 mL of the amylase solution into each of three small test tubes. Put these
into the tem"erature bath containing the starch solutions from ste" 2.
?. #llo' the solutions to remain in the tem"erature bath for about @%10 minutes to
eDuilibrate.
@. Carry out the ste"s ?%B in P#R, in seDuence for the solutions that bu>ered at "H @3
B3 E.
CC
P#R, . E>ect of Metal ons on En)yme #cti-ity
n this "art you 'ill test the e>ect of a metal ion on en)yme acti-ity3 Cu
2U
3 2e
3U
3 `n
2U
or
other transition metal ions or hea-y metal ions may be tested.
1. Label a test tube 'ith the identity of the metal ion in the salt solution you 'ill use for
this "art.
2. nto each test tube3 "lace 2.@ mL of bu>ered starch solution and 2.@ mL of the metal ion
solution. Place these into room tem"erature bath or the ice bath3 de"ending on 'hich
one ga-e better results in "art .
3. Place 1mL of the amylase solution into each of t'o small test tubes. Put them into the
tem"erature bath containing the metal ion solution.
?. #llo' the solutions to remain in the tem"erature bath for about @%10 minutes to
eDuilibrate.
@. Carry out the ste"s ?%B in P#R, in seDuence for the solutions containing the metal ion.
CE
RE02RT SHEET,Amylase" The Acti*ity of an En%yme
E0
2or each section3 record the colors of the starchFen)yme solution in the "resence of
iodine.
P#R, . E>ect of ,em"erature on En)yme #cti-ity
,ME
Kminutes:
0
o
C Room ,em"erature 100
o
C
0
1
2
?
A
C
10
Part . E>ect of "H on En)yme #cti-ity
,em"erature IIIIIIIIIIIIIIIIII
,ime
Kminutes: "H S@ "H SB "HSE
0
1
2
?
A
C
10
P#R, . E>ect of Metal ons on En)yme #cti-ity
E1
,em"erature IIIIIIIIIIIIIIIIIIIIII
,ime Kminutes:
Metal on S IIIIIIIIIIIII
0
1
2
?
A
C
10
C!=CL.S!=S
1. Gi-e a clear e$"lanation of the e>ect of tem"erature on the acti-ity of amylase.
Consult yoir class notes ideas.
2. Gi-e a clear e$"lanation of the e>ect of "H on the acti-ity of amylase. Consult
your class notes for ideas.
3. Gi-e a clear e$"lanation of the e>ect of the metal ion on the acti-ity of amylase.
E2
:nteraction of
1' .ight with
?atter
,o understand the factors that a>ect ho' certain ty"es of matter interact 'ith ./
light
,o gain a rough understanding of ho' a s"ectro"hotometer 'or*s.
,o be able to dra' conclusions about "rotecting oneTs body from ./ e$"osure.

:ntroduction
5ecalling what we have learned about light)
=igure @# We can thin energy as a wave.
'he distance between two e(uivalent points
on the wave is the wavelength .).
!ur sun emits the energy that sustains life on earth as 'e *no' it. f 'e thin* about
the energy as a wa*e3 as illustrated in 2igure 13 'e can relate the 'a-e and its associated
energy3 as sho'n in EDuation 1+
K1: E S hc F
E stands for energy. ,he energy is "ro"ortional to Planc*Ts constant Kh: and the s"eed of
light Kc3 also a constant:3 'hile it is in-ersely "ro"ortional to the wa*elength K3 lambda is
the symbol for 'a-elength: of the energy. ;hen the 'a-elength increases3 the energy
decreases3 and vice versa.
2igure 2 sho's a common diagram of an electromagnetic s"ectrum. 6ou can see
that radio and T' wa*es are made%u" of relati-ely long 'a-elengths3 and so their energy
is relati-ely lo'. ,hey "ass harmlessly around us all the time. ?icrowa*es ha-e shorter
E3
'a-elengths and ha-e higher energy. Micro'a-es ma*e 'ater molecules mo-e3 heating u"
the food you "lace in the micro'a-e o-en. :nfrared wa*es can be felt as heat3 'ith still
greater energy. ;e commonly thin* of *isible wa*es as Glight.H /isible 'a-es cause
molecules in your retina to change energy states. ,he change gets transmitted through
your o"tic ner-e by chemical signals to your brain3 'here it is inter"reted as -ision. /isible
'a-es ha-e 'a-elength of roughly B00%?00 nm3 or about the si)e of a li-ing cell5 hence3
you are able to see cells in a light microsco"e due to -isible light "assing through or
bouncing o> them. H,rays are smaller still5 one the order of the si)e of atoms. # 'a-e 'ith
a 'a-elength this small is -ery high energy_ R%rays "ass right through most organic
molecules. ,his is 'hy 'e can use them medically to see bones and other structures that
contain metals. #t the same time3 great care must be ta*en to a-oid unnecessary e$"osure
or the 7=# in cells can be damaged and cause tumors. /amma,rays K%rays: ha-e the
shortest 'a-elength3 and highest energy3 of all. Short%term %ray e$"osure is used to treat
"re%"ac*aged meats. ,he energy "asses through into the meat and *ills microbes3 thereby
e$tending the shelf life of the "roduct. ;hat can *ill microbes can *ill any cell3 thus %rays
are -ery dangerous. 2ortunately3 'e ha-e a "rotecti-e magnetic 8eld around our "lanet.
;ithout it3 life as 'e *no' it could not e$ist.
E?
=igure 2# +omparing
wavelengths .) of energy in
the electromagnetic
spectrum.
!ur e$"eriments 'ill focus on ultra*iolet light. .ltra-iolet light K./ light: has
'a-elengths from about ?00 nm3 do'n to about a nanometer. ./ light is subcategori)ed
by 'a-elength3 as sho'n in ,able 1. ,his si)e of these 'a-elenghts is on the order of that
of molecules3 and so it can interact 'ith certain functional grou"s of organic molecules. f
the organic molecule is in a li-ing organism3 changes in the molecule can ta*e "lace so that
it no longer functions "ro"erly. ;hen ./ light hits the 7=# in our s*in cells3 changes can
ta*e "lace in the chemical structure such that it no longer base "airs "ro"erly. 7uring
re"lication3 im"ro"er base%"airing can cause a mutation. f the mutation is in a region
needed for cell sur-i-al3 the cell can die "rematurely. f that mutation is in one of the many
genes that control cell gro'th3 a cancerous tumor can form.

Cells ha-e built%in mechanisms that
"rotect us from ./%induced tumor
formation. ,here are re"air en)ymes that
constantly scan our 7=# for changes in
molecular structure. ;hen found3 the
damaged "iece of 7=# is cut out and
re"laced 'ith a good "iece. #t the same
time3 'hen cell damage due to ./ light
e$"osure occurs3 our bodies "roduce a natural sun bloc* called melanin. t is the bro'n
"igment in our s*in that sho's%u" as a Wtan.T So3 a suntan is the bodyTs res"onse to cell
damage in order to try to "re-ent cancer. Premature cell death also causes the s*in to
thic*en and 'rin*le. !b-iously3 these mechanisms do not al'ays 'or*. Liberal use of sun
bloc* is essential for "eo"le of all s*in ty"es to "re-ent cell death and cancer.
&ight in the laboratory) .this section is adapted from e!periments written by <r. ,meric
Schult: for the +hemistry for the Sciences 2 lab manual/
Matter that a""ears colored is "referentially absorbing di>erent 'a-elengths of
-isible light. 6our eyes detect the com"ilation of all of the re+ected and#or transmitted
colors P the obser*ed color. #s laboratory scientists3 'e are interested in the color
absorbed3 'hich is the com"lement of the color obser-ed. ,able 1 sho's the correlation
bet'een absorbed and obser-ed -isible light. 2or e$am"le3 tree lea-es that a""ear green
to our eyes do so because red and "ur"le light is absorbed5 the green bounces o> the
lea-es3 enters our eyes3 and triggers the chemicals in the retina to signify GgreenH to the
brain.
,able 1+ Colors of /isible Light
;a-elength of Light #bsorbed Knm:
3C0 ?20 ??0 ?B0 @00 @30 @A0 @E0 A30 AC0 BC0
-
i
o
l
e
t
-
i
o
l
e
t
%
b
l
u
e
b
l
u
e
b
l
u
e
%
g
r
e
e
n
g
r
e
e
n
g
r
e
e
n
%
y
e
l
l
o
'
y
e
l
l
o
'
o
r
a
n
g
e
r
e
d
"
u
r
"
l
e
#bsorbed Color
!bser-ed Color
g
r
e
e
n
%
y
e
l
l
o
'
y
e
l
l
o
'
o
r
a
n
g
e
r
e
d
"
u
r
"
l
e
-
i
o
l
e
t
-
i
o
l
e
t
%
b
l
u
e
b
l
u
e
b
l
u
e
%
g
r
e
e
n
g
r
e
e
n
E@
,able 1% Subcategories of ./ light.
=ame ;a-elength Range
Knm:
./# Kblac* light: ?00%320
./& 300%2C0
./C Kgermicidal: &elo' 2C0
# spectrophotometer is an instrument that can measure the amount of light
GabsorbedH by matter that is dissol-ed in solution. # bare%boned schematic is sho'n in
2igure 3. !ur eyes only detects a -ery limited range of 'a-elengths. # s"ectro"hotometer
can detect any 'a-elengths it is designed to detect. ,here are ./%/isible
s"ectro"hotometers that detect from 1E0%C00 nm. ,here are R s"ectrometers3 and so
forth. n a s"ectrometer3 the light source emits 'a-es that are selected by a
monochometer. ,he chosen range of light "asses through the sam"le. Some of the light is
absorbed and does not reach the detector. ,he instrument *no's ho' much light 'as sent
into the sam"le and com"ares that to the amount that is transmitted. ,he di>erence is the
absorbance. # substance 'ill ha-e a di>erent absorbance3 de"ending u"on the
'a-elength.
Light
source
'a-elength
selector
Kmonochromator:

Sam
sam"le

Light detector
=igure A# 'he set#up for a spectrophotometer.
# scanning s"ectrometer3 as the name suggests3 can scan through lots of
'a-elengths and "roduces 'hat is called a spectrum K2igure 2:. \s"ectra is "lural for
s"ectrum]. ,here are t'o im"ortant numbers that you 'ill get from the scans of your
sam"les. ,he 8rst number is called the absorbance ma$imum and is gi-en the symbol ma$.
,he symbol Klambda: is uni-ersal in science as a designation for 'a-elength5 ma$ is
therefore the 'a-elength at 'hich a dissol-ed s"ecies interacts best 'ith light Kas sho'n
by a "ea* in absorbance:5 ma$ in 2igure ? is about @2@ nm. # 'ord about units for
'a-elength. ;a-elengths of di>erent *inds of electromagnetic radiation Ke-erything form
radio 'a-es to R%rays: can go from being -ery short to -ery long. ,he most con-enient
units to use for electromagnetic radiation in the -isible and the surrounding ultra-iolet and
infrared regions are in nanometers K10
%E
m:.

t is im"ortant for you to
distinguish bet'een 'hat ty"e of light
interacts 'ith matter Kwa*elength: and
ho' much light interacts 'ith matter
Kabsorbance:. ,a*e a loo* the 2igure 3
belo' and its labels K=ote+ although this is
a s"ectrum of -isible 'a-elengths
'hereas 'e 'ill be loo*ing at ./
'a-elengths3 the s"ectra are inter"reted
the same 'ay:.
,he second number is called the
absorbance. ,his is the Duantity that 'e
ha-e been studying. t is a measure of the
intensity of the color. 6ou *no' the factors
that a>ect this -alue. #bsorbance by
EA
=igure B# 4n absorbance spectrum for a
sample with color.
de8nition has no units. ,his means that the units selected for the factors that determine
absorbance ha-e to cancel.
Matter has a 'a-elength or 'a-elengths of light that it interacts 'ith Kabsorbs: best.
,hese 'a-elengths are called absorbance ma&ima and are labeled as ma&.KGlambda
ma$H:. !ne 'a-elength can be the -ery best but the 'a-elengths around this ma$imum
are also absorbed Duite 'ell by the molecule. ;e call this set of 'a-elengths an
absorbance band.

EB
Pre%Lab #ssignment
Scientists ha-e disco-ered many molecules that absorb ./ light and are essentially
harmless 'hen a""lied to human s*in. Before you come to lab3 use the internet
K;i*i"edia is fairly trust'orthy in this case or you can use an image search in Google: to
loo*%u" and record the structures of each of these com"ounds in your lab noteboo*+
"%#minoben)oic acid KP#&#:
#-oben)one
Cino$ate
7io$yben)one
Homosalate
Methyl anthranilate
!ctocrylene
!ctyl metho$ycinnamate K!ctino$ate:
!ctyl salicylate K!ctisalate:
!$yben)one
Padimate !
Phenylben)imida)ole sulfonic acid KEnsuli)ole:
Sulisoben)one
,itanium dio$ide
,rolamine salicylate
`inc o$ide
E$"erimental
6our instructor 'ill ha-e set%u" the Genesys s"ectro"hotometer so that it 'ill scan from
200%?00 nm.
&aseline scan of air+ the purpose of this part of the e!periment is to mae a control
scan so that atmospheric conditions do not a0ect your results.
1. Ensure the instrument sam"le com"artment is em"ty.
2. Shut the door and "ress the Gne' baselineH *ey. ,he instrument 'ill ma*e all *inds
of rude noises3 mo-e u" to the 8rst "osition3 then begin to scan.
3. 7o not o"en the sam"le com"artment until the baseline is com"leted.
?. !nce the baseline is com"leted3 the instrument 'ill return the sam"le holder to the
O2 "osition.
./ scan of Duart)+
1. Record your -isual obser-ations about the Duart) cu*et. K# cu-et is a container to
hold liDuid sam"les for s"ectrometry. ;e can only use it is it does not interfere by
absorbing the light 'e are studying.: Clean it 'ith a Qim'i"e. ;e do not 'ant
8nger"rints scattering the light_
2. Place the cu-et in the O2 "osition of the sam"le holder.
3. Shut the door3 then ma*e sure the sam"le "osition indicator in on G2.H
?. Press the Gne' scanH or GscanH *ey.
@. 7o not o"en the sam"le com"artment until the scan is com"leted.
A. Print a co"y of the scan for each lab "artner to ta"e in his or her boo*. ,a*e care to
not ta"e o-er the "rint or the adhesi-e in the ta"e 'ill dissol-e it.
EC
B. 7escribe the sha"e of the s"ectrum in your noteboo*. 2or e$am"le 'e can describe
2igure ?+ #t B00 nm3 the absorbance is )ero and then it increases to a ma$imum at
around @2@ nm. 2inally3 the absorbance decreases to around 0.1 at ?00 nm.
7oes the Duart) absorb any 'a-elengths of ./ lightN f so3 'hat is the ma$N
s this ./#3 ./&3 ./C3 or all threeN f the Duart) does not absorb ./ light3 it
is said to be G1' transparent.H f the Duart) is ./ trans"arent3 'e could
use it as a holder for liDuid sam"les and it 'ould not interfere 'ith our
s"ectra.
./ scan of glass+
1. Record your -isual obser-ations about the "iece of 'indo' glass. Clean it 'ith a
Qim'i"e. ;e do not 'ant 8nger"rints scattering the light_
2. Secure the "iece of glass into the O2 "osition of the sam"le holder.
3. Shut the door3 then ma*e sure the sam"le "osition indicator in on G2.H
?. Press the Gne' scanH or GscanH *ey.
@. 7o not o"en the sam"le com"artment until the scan is com"leted.
A. Print a co"y of the scan for each lab "artner to ta"e in his or her boo*. ,a*e care to
not ta"e o-er the "rint or the adhesi-e in the ta"e 'ill dissol-e it
B. 7escribe the sha"e of the s"ectrum in your noteboo*.
7oes the glass absorb any 'a-elengths of ./ lightN f so3 'hat is the ma$N s
this ./#3 ./&3 ./C3 or all threeN
./ scan of 'ater+
1. 2ill the Duart) cu-et 'ith distilled 'ater. ,a*e care to "re-ent air bubbles in your
sam"le.
2. Record your -isual obser-ations. Clean it 'ith a Qim'i"e. ;e do not 'ant
?. Shut the door3 then ma*e sure the sam"le "osition indicator in on G2.H
@. Press the Gne' scanH or GscanH *ey.
A. 7o not o"en the sam"le com"artment until the scan is com"leted.
B. Print a co"y of the scan for each lab "artner to ta"e in his or her boo*. ,a*e care to
C. 7escribe the sha"e of the s"ectrum in your noteboo*.
7oes the 'ater absorb any 'a-elengths of ./ lightN f so3 'hat is the ma$N
s this ./#3 ./&3 ./C3 or all threeN
./ scan of 7=#+
1. 2ill one Duart) cu-et 'ith distilled 'ater and one Duart) cu-et 'ith 7=# solution.
,a*e care to "re-ent air bubbles in your sam"les.
2. Record your -isual obser-ations about the 7=# solution in the cu-et. Clean each
'ith a Qim'i"e. ;e do not 'ant 8nger"rints scattering the light_
3. Put the cu-et 'ith 'ater in the 8rst "osition and the cu-et 'ith 7=# solution in the
second "osition.
?. Shut the door and "ress the Gne' baselineH *ey. ,he instrument 'ill ma*e all *inds
of rude noises3 mo-e u" to the 8rst "osition3 then begin to scan.
@. 7o not o"en the sam"le com"artment until the baseline is com"leted.
EE
A. !nce the baseline is com"leted3 the instrument 'ill return the sam"le holder to the
original "osition.
B. Ma*e sure the sam"le "osition indicator in on G2.H
C. Press the Gne' scanH or GscanH *ey.
E. 7o not o"en the sam"le com"artment until the scan is com"leted.
10. Print a co"y of the scan for each lab "artner to ta"e in his or her boo*. ,a*e care to
11. 7escribe the sha"e of the s"ectrum in your noteboo*.
7oes the 7=# absorb any 'a-elengths of ./ lightN f so3 'hat is the ma$N s
this ./#3 ./&3 ./C3 or all threeN
./ scan of sunscreen+
1. 7ry the Duart) cu-ets. =ear the to" of one of them3 'rite a small letter WST so that
you can distinguish the cu-ets.
2. Record your -isual obser-ations. Clean it 'ith a Qim'i"e. ;e do not 'ant
?. Shut the door3 then ma*e sure the sam"le "osition indicator in on G2.H
@. Press the Gne' scanH or GscanH *ey.
A. 7o not o"en the sam"le com"artment until the scan is com"leted.
B. Print a co"y of the scan for each lab "artner to ta"e in his or her boo*. ,a*e care to
C. 7escribe the sha"e of the s"ectrum in your noteboo*.
7oes the sunscreen absorb any 'a-elengths of ./ lightN f so3 'hat is the
ma$N s this ./#3 ./&3 ./C3 or all threeN
100
101
RE02RT SHEET,:nteraction of 1' .ight with ?atter
. Com"lete the table of results. ndicate if the substance tested absorbs ./#3 ./&3 or ./C.
2or the sunscreen3 list the name of the acti-e ingredient. Gather a fe' results from your
labmates.
Substance
Absorbs
1'AN
Absorbs 1'BN Absorbs 1'CN
Luart)
Glass
;ater
7=#
#ns'er the follo'ing. Include your scienti-c evidence to support your answer
where appropriate.
. f the 'indo's of your home or car 'ere made of Duart)3 'ould they "rotect your from
./ lightN
. 7o standard glass 'indo's "rotect your from ./ lightN
/. 7oes being under 'ater Kas 'hen s'imming: "rotect you from ./ lightN
102
/. ;hat common functional grou" is found in the acti-e ingredients of sunscreensN
/. &ased u"on your 8ndings for Duestion /3 'hat functional grou" of 7=# is li*ely to
interact 'ith ./ lightN
/. ;hat "ur"ose does this functional grou" in 7=# ser-eN
/. Ho' could ./ damage to this functional grou" cause cancerN
103

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