Carbohydrate reaction

COMPLETE REPORT ORGANIC CHEMISTRY 2

HERMIN HARDYANTI UTAMI (111 304 0202)

CHEMISTRY DEPARTMENT MATHEMATIC AND SCIENCE FACULTY STATE UNIVERSITY OF MAKASSAR 2012

RATIFICATION PAGE The complete report of Organic Chemistry II with the title of “Carbohydrate reaction” which made by : Name

: Hermin Hardyanti Utami

ID

: 111 304 0202

Group

: VII

Class

: ICP B

Have been checked by assistant and assistant coordinator. So, this report is accepted.

Makkassar, December Assistant Coordinator

Assistant

Fandi Ahmad, S.Pd.

Nurhajawarsi, S.Pd

Known By, Responsibility Lecturer

Dra. Hj. Ramdani, M.Si.

2012

A. Tittle of Experiment Carbohydrate reaction B. Purposes At the last experiment, student can be understand about : 1. Differenciate between reducing sugar with nonreducing sugar. 2. Differenciate

between disaccharide that has the aldehyde group

(hemiacetal) or ketone (hemiketal) free and non free. C. Background In theleaf of a plant, the simple compounds carbondioxyde and water are combined to rofm sugar (+) glucose. This process known as photosynthesis requires catalyst be green coloring matter chlorophyll and requires energy in the form light. Thousand of (+) glucose molecules can then be combined to form the much larger molecules of cellulose which constitute to supporting frame work of the plant (+). Glucose molecules can also be combined which is then stored in the seeds to serve as food for a new grawing plant. Positive glucose, cellulose, starch and glycogen all belong to the class of organic compounds known as carbohydrates. Carbohydrate containing in grain or feed of it to animals to be converted into meat and fat which we then eat. We build houses and furniture from cellulose in the form of wood. Rhus carbohydrates quite literally provide us with the necessary of life food, clothing and shelter (Morison and Boyd,2001:1070) According to McMurry (2008:687) classification of carbohydrate is : 1. Simple vs complex: a. Simple carbohydrates can’t be hydrolyzed to smaller units. b. Complex carbohydrates are made up of two or more simple sugar linked to sugar. 

A disaccharide is composed of two monosaccharides.



A polysaccharide is composed of three or more monosaccharides.

2. Aldose vs ketose: a. A monosaccharide with an aldehyde carbonyl group is an aldose. b. A monosaccharide with a ketone carbonyl group is a ketose.

3. Tri-,tetr-,pent-,etc-. indicate the number of carbons in the monosaccharide. According to Carrey (2000:998-999) characteristic properties of an aldehyde function is its sensitivity to oxidation. A solution of copper (II) sulphic as its citrate complex (benedict’s reagent) is comaple of oxidation aliphatic aldehydes to the corresponding carbocxylic acid: O R

C

O H + 2Cu2+

R

C

O-

+

Cu2O + 3H2O

The formation of a red precipitate of copper (II) oxide by reduction of Cu(II) is taken as positive copper test with benedict’s reagent are termed reducing sugar. Aldose are reducing sugar since they process an aldehyde function on their open chain form. Its also to ketones are reducing sugar. Under the conditions of the test ketones equilibrate with aldoses by way of enediol intermediates and the aldose are oxidated by the reagent.

According to Myamus (2006) many student have been investigating carbohydrate supplementation in relation to time of investigation frequency of investigation and concentration of the glucose solution. Although carbohydrate (glucose) is extremely important to fatty acid oxidation. Glucose then plays an important role for muscle performance especially during relatively long and intense activities. Knowing that carbohydrate is the oxidizable fuel during high intensive exercise those athletes who train intensely or complete for consecutive days without consuming adequate amounts of carbohydrates are not able mavimante glycolen storage and consequently are unable to perform at their best. According to Wissmann (2006:29) carbohydrate metabolism. Prior the examining gender related differences in carbohydrate in carbohydrates loading. Gender of different between carbohydrate metabolism must be eamined. There appears to be no difference between genders in basal level of muscle glycogen skeletal muscle in hexosinase. In addition, there have yet to be any studies conducted assessing different in glycogen synthase activity or brancing enzyme.

Knowledge of change of these enzyme could assist in the understanding of gender.

The general testing of carbohydrate that simple is molisch test. This reaction occur tof the color change as the influence of mixture of one furan derivative that form fugar reaction with acid and α-naphtol. Disaccharide has the aldehyde group (hemiacetal) or ketone (hemiketal) free, like the lactose occure in the homogeneous reaction. The melting point of crystal and formation osazone time is specific, means can be used as identification or derivative of sugar kind (Tim Dosen,2012:10-11). D. Chemical and Equipment 1. Chemical a. Aquadest (H2O) b. Ethanol (C2H5OH) solution c. Sucrose (C12H22O11)solution d. Fructose (C6H12O6) solution e. Lactose (C12H22O11) solution f. Glucose (C6H12O6) solution g. Sample sugar solution h. α-naphtol solution i. concentrated sulphuric acid (H2SO4) j. Silver nitric (AgNO3) solution k. Ammonium hydroxide (NH4OH) solution l. Benedict solution m. Phenilhydrazin (NH2NHPh) solution n. Matches o. Hydrochloric acid (HCl) 10% solution p. Sodium hydroxide (NaOH) 10% solution q. Litmus paper r. Universal indicator

s. Cotton t. Ice / cool water 2. Equipment a. Big test tube

(10 pieces)

b. Small test tube

(10 pieces)

c. Test tube rack

(2 pieces )

d. Stir bar

(3 pieces )

e. Dropping pipette

(14 pieces)

f. Beaker glass 500 ml

(2 pieces )

g. Graduated cylinder 10 ml

(3 pieces )

h. Bunsen burner

(2 pieces )

i. Triangle

(2 pieces )

j. Asbestoz Gauze

(2 pieces )

k. Spray flask

(1 piece )

l. Soft cloth and rough cloth

(2 pieces)

E. Work Procedure 1. Molisch test a. 2 ml of sugar solution (sucrose, glucose, fructose, lactose, sample) was filled into the small test tube. b. Each was added by 3 drops of α-naphtol. c. Each was added by 3 drops of concentrated H2SO4. d. Each was observed the occur change.

2. Tollens test a. Tolens reactant test was made by mix of: 

1 ml of AgNO3 5%



10 drops of NH4OH 2%

b. It was react to sugar solution (sucrose, fructose, glucose, lactose, sample) with 5 drops of each solution into small test tube. c. Each was observed the occur change. 3. Benedict test a. 5 ml of benedict solution was filled into big test tube. b. Each was added with sugar solution (sucrose, fructose, glucose, lactose, sample). c. Each was heated some minutes until change occur. d. Each was let stand until occur the precipitate. 4. Osazone test a. 10 ml of sugar solution (sucrose, fructose, glucose, lactose, sample) was filled into big test tube. b. Each was added by 5 ml of phenilhydrazin until form two layers. c. Each was shaked and and closed it with cotton. d. Each was heated until 30th minutes. e. Each was let stand the first in room temperature and the second at cold water. f. Each was observed the occur change. 5. Sucrose hydrolysis by acid a. 10 ml of sucrose solution was filled into big test tube. b. It was added by 10 ml of HCl 10%. c. It was heated until 50th minutes. d. It was checked with blue litmus paper. e. If the litmus paper is change to red, it was added by NaOH 10% until the pH is neutral. f. It was compared with benedict and took 10 drops. g. It was added by 5 ml of benedict. h. It was observed the occur change.

F. Observation Result No.

Observation

Result

Molisch test : 1.

Sample, sucrose, lactose, fructose, glucose was filled into test tube.

2.

It was added by α-naphtol.

3.

It was took 3 drops of sugar

-

2 ml of sugar solution.

-

3 drops of α-naphtol,

-

Not dissolve mixture

-

Separate :

solution. 4.

It was added by concentrated H2SO4 Tollens test :

1.

Sample solution was added by tollens reactant

2. 3. 4.

Sucrose solution was added by tollens reactant



Colorless solution

Fructose solution was added by



Purple ring

tollens reactant



Black solution

Glucose solution was added by tollens reactant

5.

Lactose solution was added by tollens reactant -

5 drops of sample, produce brownish grey solution.

-

5 drops of sucrose, produce brownish grey solution.

-

5 drops of fructose, produce silver mirror precipitate.

-

5 drops of fructose, produce silver mirror precipitate.

-

5 drops of fructose, produce

brownish grey solution. Benedict test : 1.

2.

Benedict solution (5 ml) was added by sample solution (10

-

Blue solution

drops) and let stand it.

-

Blue solution

Benedict solution (5 ml) was

-

Orange solution

added by glucose solution (10

-

Red precipitate

Benedict solution (5 ml) was

-

Dark red solution

added by lactose solution (10

-

Red precipitate

Benedict solution (5 ml) was

-

Dark red solution

added by sucrose solution (10

-

Red precipitate

-

Blue solution

-

Blue solution

-

Separate :

Benedict solution (5 ml) was added by fructose solution (10 drops) and let stand it.

3.

drops) and let stand it. 4.

drops) and let stand it. 5.

drops) and let stand it.

Osazone test : 1.

Sample solution -

10 ml of sample solution was added

by

5

ml

of

phenilhydrazin. -

-

It was shaked th

It was heated 30 minutes

It was let stand in room temperature and cold down in



Transparent solution



Yellow solution

-

Turbidity yellow solution

-

Separate :

-



Yellow oily solution



Yellow solution

Separate : 

Transparent solution



cold water 2.

Fructose solution -

10 ml of fructose solution was added

by

5

ml

-

of

phenilhydrazin. -

-

3.

It was shaked th

It was heated 30 minutes

It was let stand in room

Transparent solution



Yellow solution

-

Turbidity yellow solution

-

Separate :

-



Yellow oily solution



Yellow solution

Separate : Transparent solution

cold water



Orange solution

-

10 ml of glucose solution was by

5

ml

of

phenilhydrazin. -

It was shaked

-

It was heated 30th minutes

It was let stand in room

by

ml

phenilhydrazin. -

It was shaked

-

It was heated 30th minutes

of



Yellow solution

Separate :

-

-

5

Transparent solution

-

10 ml of lactose solution was added



Turbidity yellow solution

cold water Lactose solution

Separate :

-

temperature and cold down in

-





added

-

Separate :

temperature and cold down in

Glucose solution -

4.

Yellow solution



Yellow oily solution



Yellow solution

Separate : 

Transparent solution



Yellow solution

Separate : 

Transparent solution



Yellow solution

-

Turbidity yellow solution

-

Separate : 

Yellow oily solution

 -

Yellow solution

Separate :

It was let stand in room



White solution

temperature and cold down in



Yellow solution

cold water 5.

Sucrose solution -

-

10 ml of sucrose solution was



Transparent solution

added



Yellow solution

by

5

ml

of

phenilhydrazin. -

It was shaked

-

Turbidity yellow solution

-

Separate :

th

It was heated 30 minutes

-

Separate :

It was let stand in room temperature and cold down in cold water



Yellow oily solution



Yellow solution

Separate : 

Yellow solution



Brownish red solution

Sucrose hydrolysis by acid : 1.

10 ml of sucrose was added by 10

-

Orange solution

ml HCl 10% and heated 50th minutes. 2.

It was checked by blue litmus

-

Acid pH

3.

It was neutralized by some drops

-

Neutral pH

-

Same

of NaOH 10% 4.

It was compared with benedict, took, added by 5 ml of benedict

color

with

benedict

namely blue solution.

G. Discussion 1. Molisch Test This test has function to know about carbohydrate sample. Molisch reagent reacted to dissolving α-naphtol into alcohol. The positive test showed by purple ring forming. This test there are sample, sucrose, glucose, lactose, and

fructose that was added by α-naphtol, then added by concentrated H2SO4 with careful to forming the purple ring. This occur because carbohydrate was hydrolyzed by H2SO4 to monosaccharide and occurring hydrated to fulfular or hydroxy methyl fulfular and will be condensation to forming complex that purple color. As function of H2SO4 is as dehydrator or catalyst and α-naphtol as reagent to condensation react. The fifth sample showing the positive that contain carbohydrate. The difference color intensity that occur because sugar source from monosaccharide and disaccharide. As for the reaction is : CHO H

C

HO C

OH H

H2SO4

C

CH

3H2O

C

C

H

C

OH

H

C

OH

+

2

HOCH O COOH

CH2OH Glucose

α-naphtol O

C

CH

C

C

HOCH

O CH2

O+ O

HO3S OH SO3H Purple ring

CHO H

C

O

HO C

H

H2SO4

C

CH

3H2O

C

C

H

C

OH

H

C

OH

+

HOCH O COOH

CH2OH Fructose

α-naphtol OH

C

CH

C

C

HOCH

O CH2

O+ O

HO3S OH

SO3H

Purple ring CH2OH

CH2OH

H HO

O OH

H

H

OH

O OH O

O

O

Lactose

+2

2

H2SO4 3H2O

H

H OH

OH H

OH

HC

CH

C

C

O C

HOH2C O

OH

C

C

C

C

HOH2C

CH2

O+

O

SO3H OH

HO3S

CH2OH H HO

O OH

H

H

OH

CH2OH

O OH O

O

O

H

H OH

Sucrose

+2

H2SO4 3H2O

OH

HC C HOH2C O

CH C

O C OH

C

C

C

C

HOH2C

CH2

O+

O

SO3H OH

HO3S

Sample solution

+2

HC

O

C

CH C

HOH2C O

C

H2SO4 -H2O

complex compound purple OH

2. Tollens Test Tollens test has function to identificating the aldehyde or hemiacetal group that will be oxidated by tollens reagent. It positive test showing the silver nitric form as mark that there is carbohydrate recducing. In this experiment, the solution that produce positive is glucose and fructose, but lactose, sucrose and sample is negative result. This is not same with theory, because lactose is reducing sugar but sucrose was same with the theory is not reducing sugar because bonding of it chain between two sugar unit that included heamiacetal carbon atom and hemiketal carbon atom from two unit of sugar so is not easy to decompose and it structure chaining with it hemiacetal structure, so it result is negative. As for the structure is : 2AgNO2(aq)

+ 2NaOH(aq)

Ag2O(s) +

Ag2O(s) + 2NaNO3(aq) + H2O(l) + 4NH3 2AgNO3

+

4NH3

2NaNO3(aq) + H2O(l) 2Ag(NH3)2NO3 + 2NaOH

2[Ag(NH3)2]+ +

NO32-

Tollens reagent AgNH32+ ion will be oxidating the sugar compound that contain aldehyde group. So, AgNO3 and NH4OH has function to make tollens reagent 2[Ag(NH3)2]+ that function as oxidator. As for the reaction is :

C6H12O6 + 3OH + Ag(NH3)2+

CH2OH

CH2OH

O

H HO

C6H12O7- + 2Ag+ + 4NH3 + H2O Silver mirror

OH

H

H

OH

OH

H2O

OH

Ag(NH3)2 O

OH H

OH

glucose CH2OH

O

H HO

OH

CO2-

H

+

2Ag silver mirror

OH

CH2OH H HO

O OH

H

H

OH

CH2OH

O OH O

O

O

H

H

+ 2Ag(NH3)2+ + OH-

OH

Sucrose Sample solution + 2Ag(NH3)2+ +

OH-

3. Benedict test This test has function to differenciating reducing sugar and non reducing sugar based on it concentrated.it positive result forming the red brick precipitate. In this experiment, sugar solution (fructose, lactose, glucose, sucrose, and sample) was reacted by benedict and heated until forming red brick precipitate. Only sample and sucrose in this experiment that has no precipitate. Glucose can be reacted with Cu2+ because is aldose group that has free aldehyde group. Lactose consist of 2 monosaccharides but borh of chain is not using it hemiacetal carbon atom and easy to oxidated. Fructose because there is tautomeric namely ketose change to ketoenol and become to aldose group. Fructose is very fast to showing the positive result because there is reducing

accelerating and has the high molarity because there is aldehyde and free ketose in carbohydrate molecule and ketone group direct dehydrated become to fulfural, while aldehyde occurring transformation to ketose then become to fulfural. Sucrose cannot be oxidated because it has aldehyde group and free ketone group, it same with sample solution that showing the negative result so that it is include the nonreducing sugar. As for the reaction : CH2OH

O

H HO

OH

H

H

OH

H OH

+

Cu2+

+

Cu2O

+

5OH-

glucose CH2OH

O

H HO

OH

H

H

OH

CHOH HO

O C

O

OH

+

3H2O

red-brick

CH2OH

OH

H

H

OH

+

Cu2+

+

+

Cu2O

5OH-

Fructose COOH

OH

H HO

OH

H

H

OH

CH2OH

red-brick

+

3H2O

CH2OH

O

H HO

H

O

OH

OH

H

CH2OH

H O

O

O OH

+ Cu2+ + 5OH-

OH H

Sucrose CH2OH

CH2OH

H HO

O OH

H

H

OH

OH

O OH O

O

O

H

H

H

+

Cu2+

+

+

3H2O

5OH-

OH

Lactose COOH

OH

H

2

HO

OH

H

H

CH2OH

OH

sample solution

+

Cu2O red-brick

+ Cu2+ + 5OH-

4. Osazone test In this test, sugar solution that used is sucrose, lactose, glucose, fructose. The fourth of it solution was added by phenilhydrazin. According to the theory, monosaccharide react with more phenilhydrazin will be producing hablur that yellow color. Disaccharide that has aldehyde group (hemiacetal) or free ketone (hemiketal) like lactose occurring the homogen reaction. But at the last experiment there are no solution form the hablur. This case occur because the oxidating process is not perfectly. As for the reaction :

CHO H

C

HO C

OH H

H

C

OH

H

C

OH

+

H2N

NH

H2N

NH

CH2OH Glucose CHO H

C

HO C

H2N

NH

H

H

C

OH

H

C

OH

CH2OH Glucosazone CHO H

C

O

HO C

H

H

C

OH

H

C

OH

+

CH2OH Fructose CHO H

C

HO C

H2N H

NH

H

C

OH

H

C

OH

CH2OH Fructosazone O

H HO

H

O

OH

OH

H

CH2OH

H O

O

O OH

+ H2N

OH

NH

H

Sucrose CH2OH

CH2OH

H HO

O OH

H

H

OH

O OH O

O

O

H

H

OH H

+

H2N

NH

H2N

NH

OH

Lactose CH2OH

CH2OH

H HO

O OH

H

H

OH

OH OH O

O

O

H

H OH

Lactose 5. sucrose hydrolysis by acid In this experiment, sugar that will be hydrolyzed is sucrose with way of addition HCl that has function to hydrolyzing to glucose and fructose. This mixture was heated to accelerate the reaction process. After that solution then was cold down and neutralized with NaOH addition. Proving of neturalization suolution was checked by litmus paper. This mixture was added by benedict then forming red brick precipitate that marking that sucrose was hydrolyzed to glucose and fructose that has aldehyde group and ketone. As for the reaction is :

CH2OH

O

H HO

H

O

OH

OH

H

H O

O

HO

H

OH

OH

O

O OH

H

H

OH

OH

H

HO

OH

OH

O

CH2OH

H

4Cu

OH H H

CH2OH OH

H

H

OH

CH2OH H

OH

Sucrose

CH2OH H

CH2OH

O C OH

H

OH

H

+

OH

OH H

H CH2OH OH

2Cu2O red-brick

6.Unknown Sugar In molisch experiment, sample that was tested is positive namely has furfural group. Tollens test at addition obtained negative namely there is no silver mirror precipitate. This testing also producing negative result namely the sample is non reducing sugar. Osazone sample producing negative result namely the sample is non monosaccharide. H. Closing 1. Conclusion Based on the experiment that hade been done, so it may be conclude that : a. Reducing sugar is a carbohydrate that has free hemiacetal group on cyclic and aldehyde group on its opened chain or has ketone as hemiketal. b. Sucrose is named on non reducing sugar because of its bond between C1 on the glucose and C2 on fructose causes fructose does not has free hemiacetal anymore.

2. Suggestion a. Student can be more careful in doing the experiment. b. Student can me more compact with friends group.

BIBLIOGRAPHY

Furniss, B.Ss. dkk.1989. Vogel’s Textbook of Practical Organic Chemistry, Fifth Edition. England : Longman Group UK Limited. Mcmurry, Susan.2008. Study Guide and Solution Manual For McMurry, Fundamental of Organic Chemistry, Sevent Edition. USA : Thomson, Brooks / Cole. Rasyid, Muhaidah. 2009. Kimia Organik I. Makasar : Badan Penerbit UNM. Tim Dosen. 2012. Penuntun Praktikum Kimia Organik II. Makassar :Jurusan Kimia FMIPA UNM. Valderrama, Alonso Aguierre. Conformational Analysis of Thiosugar: Theoretical NMR Chemical Shifts And JH.H Coupling Constant Of 5-Thiopiranose Monosaccharides. Spain : Journal Of Carbohydrate Chemistry. Voss, Patricia, G.dkk. Inhibition of Free Cel Splicing by Saccharides that Bind Galectins and SR Proteins. USA : Journal Of Carbohydrate Chemistry. Wahjudi, dkk. Common Textbook, Edisi Revisi, Kimia Organic II. Malang : JICA.

Question Answer 1. The reaction of tollens test : CH2OH

CH2OH

O

H

HO

OH

H2O

OH

OH

H

H

OH

Ag(NH3)2

O

OH

H

OH

glucose CH2OH

O

H HO

OH

H

CO2-

+

2Ag silver mirror

OH

CH2OH

O

H

HO

OH

H

H

OH

O OH

O

O

O

CH2OH

H

H

+ 2Ag(NH3)2+ + OH-

OH

Sucrose 2. Molecule structure of sucrose : CH2OH

O

H

HO

OH

H

H

OH

O OH

O

O

Sucrose

O

H

CH2OH

H

OH

Sucrose gives negative result for tollens and benedict test because it has not free hemiacetal or aldehyde in open chain because it is anomeric atomic carbon bond in glycoside. 3.

Glucose and fructose have osazone crystal because its monosaccharide they have hemicetal and free aldehyde group.

4. Hydrolysis sucrose with acid react with phenilhydrazin produce compound because influence of enzyme interface that make of change of rotation polarizasm to left and right namely muersion sugar. 5. Five ethyl groups can filled to glucose and fructose, it means that they have hydrogen atoms that can replaced with acetyl group. CH2OH

O

H

HO

CHOH

OH

H

H

OH

O

CH2OH

OH

CH2OH

H

H

OH

Glucose

fructose

6. Five ethyl groups can filled to glucose and fructose, it means that they have hydrogen atoms that can replaced with acetyl group. CH2OH

O

H

HO

OH

H

O

H

OH

OH

O

O

O

H

CH2OH

H

OH

Sucrose 7. Sugar that contain sucrose after hydrolyze is glucose and fructose and sugar that reduction with benedict test is glucose.