POLYTECHNIC UNIVERSITY OF THE PHILIPPINES Sta. Mesa, Manila College of Arts e!

art"ent of Ps#$%olog# &SIOP ' ( )

*ritten Re!ort in

CAR&OHY RATE S

Presente+ ,#-

.ROUP /
&antang, Fati"a Olga

&on+o$, T%erese 0ristia &1%ain, Maria C%ristina Costales, Ra$%el .ongon, Rainier

INTRODUCTION A carbohydrate is an organic compound that is made up of carbon, hydrogen and oxygen in the ratio of 1:2:1. The general formula of carbohydrate is Cm(H2 !n.

FUNCTIONS OF CARBOHYDRATES "ubstrate for respiration #ntermediate in respiration $nergy stores (e.g. starch, glycogen! "tructural (e.g. cellulose, fungal %alls! Transport &ecognition of molecules outside a cell 'rea(do%n of fatty acids and pre)enting (etosis 'iological recognition processes *la)or and "%eeteners As an anticoagulant (heparin! As an antigen (contains oligosaccharides! As hormones As ra% material for industries li(e paper and textile +recursor for organic compounds

CLASSIFICATION OF CARBOHYDRATES MONOSACCHARIDES

Are %ater,soluble crystalline compounds. They are allphatic aldehydes or (etones %hich contain one carbonyl group and one or more hydroxyl groups. -ost natural monosaccharides ha)e either fi)e (pentoses! or six (hexoses! carbon atoms.

OLIGOSACCHARIDES These are relati)ely lo% molecular %eight polymers of monosaccharides (.2/! that are co)alently bonded through glycosidic lin(ages. 0isaccharides consist of t%o monomers, %hereas trisaccharides consist of three. POLYSACCHARIDES Structure The ma1ority of carbohydrates found in nature are present as polysaccharides. are high molecular %eight polymers of monosaccharides (22/!

GLYCOGEN #t is amylopectin %ith )ery short distances bet%een the branching side,chains. "tarch from plants is hydrolysed in the body to produce glucose. STARCH

#t is often produced in plants as a %ay of storing energy. #t exist in t%o forms: Amylose , is a linear polymer made up of 0,glucose units. Amylopectin , is a soluble polysaccharide and highly branched polymer of glucose found in plants. METHODOLOGY Isolation o Li!er Gl"co#en Materials$ *resh 3i)er (chic(en! TCA 1/4, 54 654 $thanol -ortar and +estle "and "odium chloride #ce Cheese cloth

PROCEDURE

PREPARATION OF PLANT STARCH

GENERAL TEST FOR CARBOHYDRATES MOLISCH%S TEST

IODINE TEST

BASED ON O&IDATION

BENEDICT%S TEST

FEHLING%S TEST

TOLLEN%S TEST

Place the sample in a test tube

Add Tollen's reagent

Heat the solution to gentle boiling

Take note of the silver mirror formed

OSAZONES TEST

'i sodium acetate and phen!lh!dra zine H"l and distilled To each of the test tubes( add the sugar solution. Draw the different osazones. water.

&arm with stirring until the solution cleans %top with cotton and mi well

Heat in a boiling water bath for #$ minutes E amine the cr!stals under the microscope.

"ool at room temperature

MOORE%S TEST

Place the sample in a test tube.

Add )a*H

Heat to boiling

)ote the color of the product

MUCIC ACID TEST

Place the sugar solution in separate test tubes Add concentrated H)*#

Heat the test tubes in a boiling water for an hour. "ool at room temperature

E amine and draw the cr!stal under the microscope

+f no cr!stals appear( let stand until the ne t period

+nduce cr!stalization b! scratching the tube with a clean stirring rod

BARFOED%S TEST

To /arfoed's reagent( add drops of the sugar solution

Place in a boiling water bath for . minutes

,emove and allow to cool spontaneousl!

After -. minutes( observe and record the results

BASED ON FURFURAL FORMATION

SELI'ANOFF%S TEST
+n a separate test tube( add each of the sugar solution to the %eliwanoff's reagent.

+mmerse in a boiling water bath.

*bserve the color changes in each tube during the first -. minutes of boiling.

,ecord the color and the time of formation for each solution tested.

RESULTS AND DISCUSSION

TEST OBSER(ATION INFERENCES REACTION

Molisc)%s Test

A deep )iolet coloration is produced at the

+resence of carbohydrates

This is due to the formation of an unstable condensation product of beta,napthol %ith furfural #odine forms coloured adsorption complexes %ith polysaccharides This is due to the formation of cuprous oxide by the reducing action of the sugar #f the saccharide is a reducing sugar, it %ill reduce Copper 8Cu9 (11! ions to Cu (1! oxide, a red precipitate #f the disaccharide is a reducing sugar, it %ill reduce Cu (11! ions to Cu (1! oxide

Io*ine Test

1unction of the layers "tarch, bluish blac( +resence of 7lycogen, reddish polysaccharide +resence of reducing sugars +resence of reducing purple A red precipitate is formed

Fe)lin#%s Test

Bene*ict%s Test

*ormation of a precipitate

green, red, or yello% sugar

Bar oe*%s Test

A deep blue color is +resence of reducing formed %ith a red ppt. settling do%n at the bottom or sides of the test tube sugars

Seli+ano %s Test A cherry red colored +resence of (etoses precipitate %ithin 5 +resence of aldoses minutes is obtained A faint red color produced Osa,one Test *ormation of beautiful yello% <eedle shaped crystals Hedgehog crystals "unflo%er shaped Moore%s Test crystals #t turns yello% to orange and finally dar( bro%n liberating the odor of Mucic Aci* Test caramel +resence of the bottom of the Tollen%s Test "pecific for galctose +resence of reducing sugar 7lucose=fructose +resence of maltose crystals of osa;one +resence of lactose

:hen reacted to the said reagent, (etoses react %ithin 2 minutes forming a cherry red condensation product Aldopentoses react slo%ly forming the colored condensation product &educing sugars forms osa;one on treating %ith phenylhydra;ine

This is due to the liberation of aldehyde %hich subse>uent polymeri;es to form a resinous substance, caramel. :hen galactose is oxidi;ed by nitric acid, insoluble mucic acid is formed

insoluble crystals in monosaccharide, tube results in elemental Ased to distinguish sil)er precipitating out of solution and also (no%n as ?sil)er mirror@. bet%een an aldehyde and a (etone.

#t is usually ammoniacal sil)er nitrate, but can also be other mixtures, as long as a>ueous diamminesil)er(#! complex is present.

HYDROLYSIS OF CARBOHYDRATES H"*rol"sis is the decomposition of a chemical compound by reaction %ith %ater, such as the dissociation of a dissol)ed salt or the catalytic con)ersion of starch to glucose. T%o types of Hydrolysis: B B Acid hydrolysis $n;ymatic hydrolysis

Aci* H"*rol"sis Acid hydrolysis is a chemical process in %hich acid is used to con)ert cellulose or starch to sugar. METHODOLOGY

CThe acid that %as used in this experiment to hydroly;e the carbohydrate is the hydrochloric acid (HCl! En,"-atic H"*rol"sis $n;ymatic hydrolysis is a process in digestion in %hich macromolecules are split from food by the en;ymatic addition of %ater. METHODOLOGY

C "ali)a %as used in this experiment to hydroly;e carbohydrates because sali)a contains en;yme that can cataly;e or hydroly;e starch into sugars, this en;yme is called Amylase. A-"lase is an en;yme that catalyses the brea(do%n of starch into sugars. Amylase is present in human sali)a, %here it begins the chemical process of digestion. *oods that contain much starch but little sugar, such as rice and potato, taste slightly s%eet as they are che%ed because amylase turns some of their starch into sugar in the mouth.

Classi ications o A-"lase$ B Al.)a/a-"lase , is found in pancreas. #t hydrolyses dietary starch into disaccharides and trisaccharides %hich are

con)erted by other en;ymes to glucose to supply the body %ith energy. B Beta/a-"lase , cuts starch into maltose units. They are present in yeasts, molds, bacteria, and plants, particularly in the seeds. #t is important in ma(ing beer and li>uor be)erages. This process is important in the digestion of starch and is also used in bre%ing, %here amylase from the s(in of seed grains is responsible -ashing!. for con)erting starch to maltose (-alting,

ANS'ERS TO 0UESTIONS$ Pre!aration of !lant star$% 2. *%at is t%e !1r!ose of grin+ing t%e li3er 4it% san+ an+ TCA5 --Glycogen is extracted from animal tissue. This tissue is actually pretty tough and to liberate the glycogen from it, the tissue has to be ruptured, broken down nicely, so that all the glycogen is available for extraction. This grinding (homogenizing process is aided by the presence of sand and T!" (Tri-!hloro "cetic acid . T!" is a higly corrosive agent and helps in rupturing the tissue. ). Li3er gl#$ogen an+ !lant star$% 4ere !re!are+ in +ifferent 4a#s. A$$o1nt for t%e +ifferen$e The starch extracted by grinding it and mix it with water then strain it using the cheesecloth, while in the extraction of glycogen, we used sand and tca before and then we strain using the cheesecloth /. Na"e ) $%e"i$al $o"!onents of star$%. --#tarch consists of two types of molecules, amylose (normally $%-&%' and amylopectin (normally (%-)%' . .eneral Test for Car,o%#+rates 2. is$1ss ea$% of "e$%anis" of a$tion. t%e tests a,o3e. S%o4 t%e

*. +,-+./ T/#T- indicates and detects the presence of polysaccharide $. 0/./-+!T1# T/#T- general test for detecting the reducing sugars &. ,#"2,./ T/#T- This is a test for reducing carbohydrate, reducing dissacharides and manymonosaccharides to be identified with the formation of osazone crystals.

3. #/4+5".,661# T/#T- This test is used to differentiate between ketoses and aldoses. 7. 8,4+#!91# T/#T- a sensitive chemical test for the presence of carbohydrates, based on the dehydration of the carbohydrate by sulfuric acid to produce an aldehyde :. 0";6,/-1# T/#T- a chemical test used for detecting the presence of monosaccharides. (. 8<!+! "!+- T/#T- specific for monosaccharide galactose ). 6/94+.G1# T/#T- 6ehling=s solution is a chemical test used to differentiate between water-soluble carbohydrate and ketone functional groups, and as a test for monosaccharides. >. 8,,;/1# T/#T- indicates the presence of a carbohydrates in a specific compound. *%. /.2?8"T+! 9?-;,4?#+#- is a process in digestion in which macromolecules are split from food by the enzymatic addition of water. **. "!+- 9?-;,4?#+#- is a chemical process in which acid is used to convert cellulose or starch to sugar ). A!!ro6i"atel# %o4 "an# ti"es faster +oes fr1$tose rea$% to gi3e a $olore+ !ro+1$t $o"!are+ 4it% t%e ot%er s1gar sa"!les5 --6ructose is the sweetest saccharides compared to other solution. "pproximately, fructose is *%x faster to reach or to give a colored product. /. ifferentiate &ene+i$t7s fro" Fe%ling7s sol1tion

--6ehling=s solution has two li@uids (one, a solution of !opper sulfate and the other made with ;ochelle salt, potassium sodium tartrate tetrahydrate and #odium hydroxide . The two parts are combined Aust before use. The reaction involves the formation of !opper (++ oxide which is reduced to !opper (+ oxide in the presence of "ldehydes (including reducing sugars .

0enedict=s solution is a single solution of !opper (++ sulfate dissolved in a #odium carbonateB#odium citrate buffer. The soluble !opper C$ ion is directly reduced to the !opper (+ oxide with reducing sugars and aliphatic aldehydes. 0enedict1 is more sensitive than 6ehling1s '. Ho4 i"!ortant is ti"e in t%e $%ara$teri8ation of t%e s1gar 1sing &arfoe+7s test -- -o not heat longer than $ minutes. 4onger heating may cause hydrolysis of the glycosidic linkage, thus breaking disaccharides to monosaccharides. 9. *%at !re$i!itate res1lts 4%en re+1$ing s1gars are %eate+ 4it% Fe%ling7s, &ene+i$ts7s an+ &arfoe+7s reagents. -- Generally, the result is brick red precipitate.

H#+rol#sis of Car,o%#+rates 2. *%at is t%e en8#"e !resent in t%e sali3a t%at %#+rol#8es t%e star$%5 -- "mylase is an enzyme that catalyses the breakdown of starch into sugars. "mylase is present in human saliva, where it begins the chemical process of digestion. ). Relate 3is$osit# %#+rol#sate 4it% t%e $o"!osition of t%e

-- The viscosity is only dependent on the difference between the actual temperature and the glass transition temperature of each starch hydrolysate. /. Offer an e6!lanation for t%e 1se of t%e +ial#8ing ,ag (( +t was used as a container of solution (carbohydrate and saliva and the bag suspend in water to undergo the process of enzymatic hydrolysis. +t also acts as a filter bag where the

starch goes out from the tube and the protein remains inside. Pa!er C%ro"atogra!%# in Un:no4n S1gars 2. In intro+1$ing t%e sa"!le, t%e s!ots s%o1l+ ,e :e!t as s"all as !ossi,le. *%#5 -- +t should be taken to keep the spot size as small as possible to keep the bands concentrated for accurate comparisons and to allow easier viewing of the tags. ). *%at is t%e $%e"i$al ,asis for t%e aniline(a$i+( o6alate s!ra# t%at lea+ to t%e $olor test -- "niline-acid-oxalate spray allows the reducing sugar1s hemiacetal carbon to expose their aldehyde. "rrangements of these kinds enable to reduce cupric to cuprous which are linked to various color reactions. /. &ase+ on t%e Rf 3al1es of t%e stan+ar+s an+ t%e 1n:no4ns, $o"!are t%e !ro+1$ts of a$i+ an+ en8#"ati$ %#+rol#sis -- The starch and glycogen are both polysaccharide and they undergo the process of hydrolysisD the polysaccharide hydrolyzed into monosaccharide which is glucose. They1re both glucose so they have an e@ual ;f values. The glucose is very soluble in water hence the ;f value of starch and glycogen are high. '. After +e3elo!ing t%e $%ro"atogra", 4%# is it ne$essar# to en$ir$le t%e s!ots !rior to t%e $al$1lation of t%e Rf 3al1e5 -- +t is necessary to encircle the spots so that it1s easier for us to know or to observe what is the exact location of the spots prior to the calculation of the ;f value because some solutions evaporates @uickly.