Bio Chem Prelims Hand Outs

7/24/2019 Bio Chem Prelims Hand Outs

1/18

Waylon FanBSP2B

EXPERIMENT 4PROCESS IN BIOCHEMICAL SYSTEM

OBJECTIVES

1. To relate the particle concept of matter to biochemical processes;2. To describe molecular motion and interaction; and3. To observe and describe processes that occur in biochemical system:diffusion, dialysis, osmosis, hydrolysis and lowering of surface tension.

REAGENTS / MATERIALS2 !a"l #$n% & 'efibrinated blood $icroscope%.1 !a"l %.( !a"l )ile *tring1 !a 2c%3 1 pacreatin $argarine1 $ +g! 3 %.- "u*% & onganisa membrane1% !a 0odine solution "hic en bouillon%.2 !inhydrin $acaroni Thermometer

PROCEDURE

A. Dialysis

1. btained one 1% cm long dialysis tubing or gastric lining longganisa membrane about2.% 2.- cm in diameter or colorless cellophane about 2- cm s4uare in si5e.6

2. 0f gastric lining to be used, tightly tie a string around its lower portion to close one end.7et the lining or dialysis tubing6 and open the other end with the stirring rod to chec ifthe pouch or sac is made.

3. 8our about 2- m of chic en bouillon previously dissolved in hot water6 into the prepared

lining pouch and tie a string around the upper potion carefully so that there will be nolea .

&. 7ash off with distilled water any spilled solution on the outside of the pouch/sac .-. 8lace the lining with its contents in a bea er and add distilled water 9ust enough to

immerse about three fourths of it. Note: The distilled water to be used must be first checked for negligible amount of Cl. To do this, test

1.0 mL of the distilled water from the large portion and add 1.0 mL of 1.0 M sil er nitrate solution. Theresult must be colorless to neglible turbidit!."

. +llow to stand for an hour at room temperature, following the set up given.

. Test the dialysate by doing the following:

a. Test !" C#l!"i$e i!%.To 1.% m of the dialysate, add 1.% m of +g! 3 solution.

bservation: There is white precipitate, there is clear li4uid

&. Test !" 'e'ti$e &!%$To 1.% m of dialysate, add 2.% m of 1% !a and - drops of %.- copper sulfate. *tir the

solution.


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Waylon FanBSP2B

bservation:7hen add w/ 1% !a it was clear, when added with %.- copper sulfateformedlavender color w/ blue precipitate.

(. Test !" a)i%! a(i$To 3.% m of the dialysate, add 1 m %.2 ninhydrin. eat in a water bath until a blue color

appears which indicates the presence of amino acid.bservation:inse the bea er and fill it with 2% m water. eat the water to the temperature of % %"

to ?%%

". +gain, add crystals of #$n%& to the still water in the same way as step 2 above..!ote the time of migration in the second along the same distance at the higher temperature.

7hat is the effect of increasing the temperature on the rate of the migration of the solidsubstance in a li4uid medium@

7hen the temperature is hot / high temp. The rate of migration is faster while if thetemperature is cold, the migration is slower.


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Waylon FanBSP2B

'raw the set up of apparatus and describe the directional movement of the colored particles.

C!)'a"e t#e "ate ! $i *si!% at $i e"e%t te)'e"at*"es. 2. % "; 1 min. &1 seconds ?% % "; &2 seconds

slow; cold temperature fast; hot temperature

C. Os)!sis

1. To each of four small test tubes, place - drops of defibrinated blood. abel thefirst test tube, A 1 as the control.

2. To the other test tubes, starting with the second, add 2% drops of the following: test tube A 2 %.1 !a"ltest tube A 3 %.( !a"ltest tube A & 2.% !a"l

3. *ha e. Bet a small amount of contents from each test tube, place on separateslides and eCamine under the microscope the appearance of the blood cells.

ow do the shape and overall picture of the cell from the three test tubes compare with that of thecontrol@

+nswer by drawing the appearance of red blood cell as seen under the microscope showing thedifference/s in their si5e and shape. 'escribe them accordingly. =se the following table.

ow the >)"Ds compare in si5e and shape

'rawings


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Waylon FanBSP2B

'escription*i5e: argest

*hape: circle withbiconcave

"olor: 1 st E >ed

2nd

E ight green 3 rd E white

*i5e: The smallest amongof them

*hape: circle withbiconcave

"olor: Fellowish white

*i5e: The second largestamong the & sample

*hape: "ircle with thebiconcave

"olor: "olorless

*i5e: *econd smallest of the & sample

*hape: "ircle with thebiconcave

"olor: colorless

Test tube A 1control6

Test tube A 2in %.1 !a"l6

Test tube A3in %.( !acl6

Test tube A&in 2 !a"l6

D. Hy$"!lysis

1. 8lace 1 piece of elbow macaroni into each of 3 small test tubes. +dd - m of pancreatin solution to the first test tube; - m of 1 !a 2c%3 to another test tube

and - m of saliva to the test tube.2. 8lace the three test tubes in a water bath with the temperature controlled at &%-%%" for strictly +, )i%*tes.

3. 8our of the solution and rinse the macaroni pieces with distilled water. 8lacethem in a watch glass e4uidistant to each other. bserve each.

&. 8lace a few drops of iodine solution on the surface of each pieces of macaroniand observe the resulting intensity of the color. $a e comparisons on the teCtureand appearance of the threated macaroni pieces after heating and with 0odinesolution.

Sa)'le )a(a"!%i Te-t*"e a te" +, )i%*tes #eati% C!l!" /it# t#e I!$i%e s!l*ti!%A1 *lightly soft/slightly viscous )lue violetA2 *oft / viscous "harcoal / slightly hardens

A3 ard / smooth "harcoal / it hardens E. L!/e"i% ! S*" a(e Te%si!%

1. +pply a thin uniform coating of margarine on one 3 slides.2. "over the margarine on one slide with bile solution, the other with %.- !a 2c%3 and the last one

with soap solution. et them stand for 3% minutes.3. 8our of the solutions and rinse each slide several times with running water very carefully. old

the slide up against the light and observe what happens.&. $a e a comparison on degree of disappearance / solubility of the coated margarine soa ed with

different solution. +ccount for observations.

$argarine G bile solution The bile solution is not soluble in margarine, the bile solution faded

$argarine G !a 2c%3 $argarine and !a 2c%3 is soluble. The !a 2c%3 solution faded

$argarine G soap solution $argarine and soap solution is slightly soluble in terms of degree of disappearance, the soap moderately faded.

SUPPLEMENTARY 0UESTION


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Waylon FanBSP2B

1. 'ifferentiate passive transport and active transport system. Bive eCamples. 8assive transport does not re4uire energy when substance move across in a cell membrane.

eCample: ydrated !a G is larger that # G and therefore tends to move easily6

2. 0s energy always associated with biochemical processes@ Hustify your answer. !o, active transport always occur against an electrical or chemical reagent

3. 'iscuss the movement of !a G and # G ions in the cell. *odium potassium eCchange pumps moves !a G out of the cells and # G into the cells. The result

is the higher concentration of !a G outside the cell.

EXPERIMENT ,GENERAL REACTIONS O1 CARBOHYDRATES

Ca"&!#y$"atesare the main energy source for the human body.

are organic molecules in which carbon, hydrogen, and oCygen bond together

+nimals including humans6 brea down carbohydrates during the process of metabolism torelease energy .

+nimals obtain carbohydrates by eating foods that contain them.

Si)'le S* a"s +ll carbohydrates are made up of units of sugar also called saccharide units6. "arbohydratesthat contain only one sugar unit monosaccharides6 or two sugar units*imple sugars are sweet in taste and are bro en down 4uic ly in the body to release energy .

Two of the most common monosaccharides are glucose and fructose. Blucose is the primaryform of sugar stored in the human body for energy .

C!)'le- Ca"&!#y$"ates

are polymers of the simple sugars.

are long chains of simple sugar units bonded together

compleC carbohydrates are often referred to as polysaccharides

Sta"(#

is the principal polysaccharide used by plants to store glucose for later use as energy .

8lants often store starch in seeds or other speciali5ed organs
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Waylon FanBSP2B

1*%(ti!%s ! Ca"&!#y$"ates"arbohydrates have several functions in cells.

They are an eCcellent source of energy for the many different activities going on in our cells.*ome carbohydrates may have a structural function.The material that ma es plants stand tall and gives wood its tough properties is a polymer form

of glucose nown as cellulose . $a e up the stored forms of energy nown as starch and glycogen .

"arbohydrates are essential for cells to communicate with each other.

They also help cells adhere to each other and the material surrounding the cells in the body.

The ability of the body to defend itself against invading microbes and the removal of foreignmaterial from the body such as the capture of dust and pollen by the mucus in our nose andthroat6 is also dependent on the properties of carbohydrates.=se in wine ma ing

M!lis(#2s Testis a chemical test for the presence of carbohydrates , based on the dehydration of thecarbohydrate by sulfuric acid to produce an furfural derivatives anion with alpha napthol to formcolored compleCes.

+ positive reaction is indicated by a purple ring forming at the interface between the acid andtest layers.

RESULT

the formation of a purple product at the interface of the two layers.

a negative test left6 and a positive test right6

ANTHRONE 3 BLUE OR GREEN COLOR

ALDEHYDE RETONE HAS REDUCING PROPERTIESOXIDI5ED AL6ALINE SOLUTIONS O1 CUPRIC SUL1ATEREDDISH BRO7N PPT. 8CUPRIC TO CUPROUS9

Be%e$i(t2s "ea e%t

is a solution of copper sulfate, sodium hydroCide, and tartaric acid. +4ueous glucose is miCed with)enedictIs reagent and heated.The reaction reduces the blue copper 006 ion to form a bric red precipitate of copper 06 oCide.)ecause of this, glucose is classified as a reducing sugar.
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Waylon FanBSP2B

RESULT

The formation of a reddish precipitate within three minutes.

>esults of )enedictIs Test with Blucose, *ucrose, and $+T0 ! < $JT+ 0" )0*$=T

)+>< J'* >J+BJ!T "=8>0" +"JT+TJ 0! 7J+# +"JT0" +"0', >J'="0!B$ ! *+"" +>0'J* ! T '0**+" +>0'J*

$ ! *+"" +>0'J*'0*+"" +>0'J*8 F*+""+ >
http://en.wikipedia.org/wiki/Solutionhttp://en.wikipedia.org/wiki/Solutionhttp://en.wikipedia.org/wiki/Aldehydehttp://en.wikipedia.org/wiki/Aldehydehttp://en.wikipedia.org/wiki/Ketonehttp://en.wikipedia.org/wiki/Ketonehttp://en.wikipedia.org/wiki/Functional_grouphttp://en.wikipedia.org/wiki/Functional_grouphttp://en.wikipedia.org/wiki/Functional_grouphttp://en.wikipedia.org/wiki/Precipitatehttp://en.wikipedia.org/wiki/Hermann_von_Fehlinghttp://en.wikipedia.org/wiki/Aldosehttp://en.wikipedia.org/wiki/Aldosehttp://en.wikipedia.org/wiki/Solutionhttp://en.wikipedia.org/wiki/Aldehydehttp://en.wikipedia.org/wiki/Ketonehttp://en.wikipedia.org/wiki/Functional_grouphttp://en.wikipedia.org/wiki/Precipitatehttp://en.wikipedia.org/wiki/Hermann_von_Fehlinghttp://en.wikipedia.org/wiki/Aldose


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T!lle%s2 "ea e%t

is usually ammoniacal silver nitrate, but can also be other things, as long as thereis an a4ueous diamminesilver 06 compleC

#$%&LT

*ilver mirror image

EXPERIMENT ,GENERAL REACTIONS O1 CARBOHYDRATES

)HJ"T0KJ*:

1. The ob9ective of this eCperiment is to be able to describe the properties and structures ofcarbohydrates in general.
http://en.wiktionary.org/wiki/aqueoushttp://en.wiktionary.org/wiki/aqueoushttp://en.wikipedia.org/wiki/Ammonia#Ammonia_as_a_ligandhttp://en.wikipedia.org/wiki/Ammonia#Ammonia_as_a_ligandhttp://en.wikipedia.org/wiki/Complex_(chemistry)http://en.wikipedia.org/wiki/Complex_(chemistry)http://en.wiktionary.org/wiki/aqueoushttp://en.wikipedia.org/wiki/Ammonia#Ammonia_as_a_ligandhttp://en.wikipedia.org/wiki/Complex_(chemistry)


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Waylon FanBSP2B

2. to differentiate reducing sugars from non reducing sugars.

$+TJ>0+ *:

1 glucose 1 sucrose

conc. *ulfuric acid 1 silver nitrate

cupric acetate 1 lactose

1 maltose 1 starch

1 )a 6 2 ! !a

phenolphtalein

! & 1 Cylose

picric acid 1 glycogen

dil. " !ylanderDs reagent

glacial acetic acid )enedictDs reagent


10/18

Waylon FanBSP2B

TEST COMPOUND OBSERVATION

Blucose G positive result6upperlayer lavender;lower dar purpleLylose G positive result6 upperlayer lavender;lower dar purple

*ucrose G positive result6upperlayer white;lower dar est purple*tarch G positive result6upper grey white;lower blac

$altose G positive result6upper cloudywhite;lower lavender


Di$ all t#e sa)'les i;e t#e sa)e "es*lt< +ll of the test compounds give positive and same result. They are all carbohydrates.

A((!*%t !" t#e "es*lts !&se";e$

+ll of them are carbohydrates that is why they all formed violet color at the 9unction upon addingmolisch reagent.

B. REACTION 7ITH+. MOORE=S TEST

a. to 1 ml of 1 glucose, add 2ml of !a , heat in a boiling water bath. 7#at(#a% es i% (!l!" a%$ !$!" #a;e y!* !&se";e$ @

0n !a , 0t was odorless and clear, turns yellowish in color, after boiling , it turns clear goldenbrown. 0t has a slight caramel odor. 0n )a 6, 0t was odorless and cloudywhite and has white precipitate. +fter boiling, it turns brownish orange and has a caramel odor.

C. REDUCING PROPERTY 8reparation of TollenDs reagent:

To -ml of 1 +g! 3 solution, add 1 drop of ! !a , +dd ammonia solution dropwise,sha ing after each addition until the precipitate initially formed is dissolved. +void adding more what isnecessary to dissolve the precipitae.


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+. T!lle%=s Test

8lace drops of 1 glucose into a clean test tube and add 2 ml of tollenDs reagent. $iC well. +llow to stand for - minutes. 7#at $i$ y!* !&se";e$. 1e#li% =s Test

a.8repare K+T0 !*

?. Nyla%$e"=s test a. miC 1- drops of 1 glucose with an e4ual amount of nylanderDs solution in a test tube.

eat for - minutes on a boiling water bath. !ote the color produced.b. >epeat the procedure on 1 Cylose, 1 sucrose, 1 starch and 1 lactose solutions.

TEST COMPOUND OBSERVATION1 glucose1 Cylose1 sucrose


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1 starch1 lactose 7#i(# (!)'!*%$ a;e t#e '!siti;e "es*lt< Blucose, Cylose and lactose gave the positive result.

7#i(# (!)'!*%$ $i$ %!t a;e t#e '!siti;e "es*lt< *ucrose and starch did not gave positive result.

4. Be%e$i(t=s test

a. to 1 ml of benedictDs reagent in a test tube, add 1ml of 1 of glucose solution. eat in aboiling water bath in 2 minutes and allow to stand. !ote the time re4uired to show agreen or red color.

b. >epeat the test on 1 solutions of Cylose, sucrose, lactose, starch and glycogen.

TJ*T " $8 =!'* )*J>K+T0 !glucoseCylose

sucrosestarchlactoseglycogen

D. HYDROLYSIS

1. 8repare the rice solution by placing a spatula of coo ed rice in a mortar and poundingusing a pestle. +dd 1% ml of distilled water. *tir

2. 8ut 2ml of rice solution in a test tube, add 1 ml of $ "l and heat in a boiling water bathfor 1- minutes to hydroly5ed.

3. !eutrali5e the resulting miCtures with about 1% drops of $ !a . Test by adding a dropof phenolphthalein. + light pin color should be observed.

&. +dd 2ml of fehlingDs reagent. eat in a water bath for 2 minutes-. 0n another test tube, miC 2ml of rice solution, 1ml of fehlingDs + and 1 ml of fehlingDs ). heat

in a water bath for 2 mnutes. 7as t#e"e a (#a% e i% (!l!"< !one Is a '!siti;e "es*lt !&tai%e$< Fes 7#at $!es it i%$i(ate< + reddish brown precipitate

EXPERIMENT @SPECI1IC GENERAL REACTIONS O1 CARBOHYDRATES

Ca"&!#y$"ates


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Waylon FanBSP2B

are classified generally according to their degree of compleCity.the free sugars such as glucose and fructose are termed monosaccharides; sucrose andmaltose, disaccharides; and the starches and celluloses, polysaccharides."arbohydrates of short chain lengths such as raffinose, stachyose and verbascose, whichare three, four and five sugar polymers respectively, are classified as oligosaccharides.

Pe%t!sesare five carbon sugars seldom found in the free state in nature. 0n plants they occur in polymeric forms and are collectively nown as pentosans.

He-!sescomprise a large group of sugars.8rincipal among these are: glucose, fructose, galactose and mannose. 7hile glucose andfructose are found free in nature, galactose and mannose occur only in combined form.The heCoses are divided into aldoses and etoses according to whether they possessaldehydic or etonic groups.

Gl*(!se is widely distributed in small amounts in fruits, plant 9uices and honey.0t is commercially produced by the acid or en5yme hydrolysis of grain and root starches.Blucose is of special interest in nutrition because it is the end product of carbohydratedigestion in all non ruminant animals including fish.

1"*(t!seis the only important etoheCose and is found in the free state alongside glucose inripening fruits and honey."ombined with glucose it forms sucrose.


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Waylon FanBSP2B

>affinose, which is the most widespread among the three, consists of one molecule ofglucose.

P!lysa((#a"i$es represent a large group of compleC carbohydrates which are condensation products ofundetermined numbers of sugar molecules.

I!$i%e test is used to test for the presence of starch . 0odine solution M iodine dissolved in an a4ueous solution of potassium iodide M reactswith starch producing a deep blue blac color

RESULTThe formation of a blue blac compleC.


Bial=s !"(i%!l=s test + test for the presence of a pentose in urine, utili5ing oracin, hydrochloric acid, and ferricchloride; a green color or green precipitate indicates pentose.

RESULT

The formation of a bluish product. +ll other colors indicate a negative result for pentoses. !otethat heCoses generally react to form green, red, or brown products.
http://en.wikipedia.org/wiki/Starchhttp://en.wikipedia.org/wiki/Starchhttp://en.wikipedia.org/wiki/Iodinehttp://en.wikipedia.org/wiki/Potassium_iodidehttp://en.wikipedia.org/wiki/Potassium_iodidehttp://en.wikipedia.org/wiki/Starchhttp://en.wikipedia.org/wiki/Starchhttp://en.wikipedia.org/wiki/Starchhttp://en.wikipedia.org/wiki/Iodinehttp://en.wikipedia.org/wiki/Potassium_iodidehttp://en.wikipedia.org/wiki/Starch


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two negative tests left, middle6 and a positive test right6

Seli/a%! =s test

+ color test helpful in the identification of etoses, which develop a red color withresorcinol in hydrochloric acid

RESULT

The formation of a red product.


EXPERIMENT @SPECI1IC REACTIONS O1 CARBOHYDRATES


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OBJECTIVES:

1. To differentiate the reactions of heCoses from pentoses, aldoses from etoses.2. To identify different tests for specific carbohydrates.3. To identify an un nown carbohydrate sample.

MATERIALS:

1 ="T *J 1 B+ +"T *J1 B =" *J 1 LF *J1 +"T *J *T+>"*="> *J * =T0 ! B F" BJ! * =T0 !)0+ I* >J+BJ!T " !". !0T>0" +"0'*J 07+! +"T *J 8 7'J>*="> *J 8 7'J> * '0=$ +"JT+TJ

PROCEDURE:A. BIAL=S ORCINOL TEST

1. 8ipet %.- ml of 1 glucose solution into a test tube and 1.% ml of rcinolDs reagent. 8lace thetest tube into a hot water bath. 7#at is t#e (!l!" !")e$ $*"i% t#e i"st , )i%*tes< +fter - minutes, you will observe changes in color, the result is a blue green solution. 2. >epeat the test on 1 fructose, 1 Cylose, and an un nown solution.

7#i(# ! t#e test (!)'!*%$s a;e a '!siti;e "es*lt< Lylose gave the positive result of having a blue green solution. T! /#i(# "!*' ! (a"&!#y$"ates $!es it &el!% < Lylose belong to the group of pentoses

two negative tests left, middle6 and a positive test right6

B. MUCIC ACID TEST


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Waylon FanBSP2B

1. 0nto & separate test tubes, place a pinch of each of galactose, glucose, lactose, and anun nown. +dd 1 ml of water and 1ml of conc. !itric acid.

2. eat the test tubes with their contents in a boiling water bath for an hour.3. "ool at room temperature. 0nduce crystal formation by scratching the tube with a clean

stirring rod. "ollect some crystal with each tube.&. JCamine with a microscope.

GALACTOSE GLUCOSE LACTOSE UN6NO7N

G6 8ositive resultformation of insolublecrystals




Base$ !% t#e ("ystals !")e$ /#at is t#e *% %!/% sa)'leepeat the test using 1 fructose, 1 sucrose, 1 starch solution

T! /#i(# "!*' ! (a"&!#y$"ates $! t#ey &el!% < Ca"&!#y$"ates G"!*' O&se";e$ (!l!" Ti)e1. glucose $onosaccharide Fellow color 2 min

2. fructose 'isaccharide >ed cherry 3 min

3.sucrose 'isaccharides >ed cherry 3 min

&. starch $onosaccharides Fellow color 22 min

7hich of the samples reacted most readily to the test@

Blucose readily reacted to the test.


D. IODINE TEST


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Waylon FanBSP2B

0n a spot plate or watch glass, separately place - drops of each of the 1 solutions of starch, glycogenand glucose. To each spot, add a drop of iodine solution. 7hat color was observed with: *tarch : bluish blac / blue solutionBlycogen : red/ brown solutionBlucose : light yellow/ colorless solution


*=88 J$J!T+>F N=J*T0 !*:

+. 7RITE THE STRUCTURAL 1ORMULA O1 GLUCOSE AND 1RUCTOSE.

Gl*(!se 1"*(t!se

>. 7RITE THE STAGES IN HYDROLYSIS O1 STARCH AND THEIR COLOR R7ACTIONS 7ITHIODINE SOLUTION.

+. gelatini5ation). li4uefaction

C. *accharification?. EXPLAIN THE 1ORMATION O1 OSA5ONE CRYSTALS AND MUCIC ACID.

The reactions served to identify the sugars by the structure of the crystals and the re4uiredtime from them upon oCidation with nitric acid heCoses produce crystals that are soluble indilute acid and 2 .

Bio Chem Prelims Hand Outs

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