Characterization of Saponifiable Lipids

April 21, 2018 | Author: Ding Regino | Category: Lipid, Fat, Fatty Acid, Membrane Active Molecules, Hydrolysis
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Biochem lab...

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Characterization of Saponiable lipids 3PSY5

Members Mercado, Eloisa Belle Pestano, Julian Terrence Regino, Paul Andrew R.  Tad,  T ad, !ristine !ristine "laire "laire  Tee,  T ee, Margarita Margarita #illanue$a, !er$in













Members Mercado, Eloisa Belle Pestano, Julian Terrence Regino, Paul Andrew R.  Tad,  T ad, !ristine !ristine "laire "laire  Tee,  T ee, Margarita Margarita #illanue$a, !er$in













%ntroduction RE&%'(, Paul Andrew R. 3PSY5

)i*ids •





)i*ids are naturall organic com*ounds t+at are insoluble in water but soluble in organic sol$ents. Animals li*ids ma-e u* most o t+e ats in t+e orm o adi*ose tissues w+ic+ +el*s in insulation and t+ermal regulation. /ats are -nown to store +eat. Plants li*ids are seen as t+e wa0

"lassi1cation o li*ids •

(*en2c+ain com*ounds wit+ *olar +eads and long non*olar tails . 4. 3. . 5.



/att acids Triacglcerols S*+ingoli*ids P+os*+oli*ids &lcoli*ids

/used2ring com*ounds . Steroids

"lassi1cation o li*ids •

(n t+e basis o w+et+er t+e will undergo +drolsis in al-aline solution 1. Saponiable lipids

4. 'onsa*oni1able li*ids

/att Acids Polar end6 t+e carbo0l grou* 'on*olar tail6 t+e +drocarbon c+ain /att acids are am*+i*at+ic  T+e usuall +a$e e$en number o carbon atoms 7 t+e +drocarbon c+ain is unbranc+ed









 Two main t*es Saturated 7 unsaturated



 Triaclglcerols  T+e 3 alco+ol grou*s in glcerol 8glcerol contains 3 +dro0l grou*s9 orms ester lin-ages wit+ att acids. /unction storage 8es*eciall in animals9





P+os*+oli*ids  T*e o li*id t+at contains a *+os*+orus atom attac+ed to t+e glcerol bac-bone  T+e are -nown to be t+e main li*id com*onent o t+e cell membrane 4 t*es o *+os*+oli*ids







. P+os*+oaclglcerols2 glcerol is t+e bac-bone 4. S*+ingomelins2 s*+ingosine instead o glcerol

&lcoli*ids •





A li*id to w+ic+ a sugar moiet is bonded A carbo+drate is bound to an alco+ol grou* o a li*id b a glcosidic lin-age "eramides are t+e *arent com*ounds or glcoli*ids

S*+ingoli*ids •



:o not contain glcerol but instead t+e contain sphingosine. /ound bot+ in *lants and animals 7 t+e are abundant in t+e ner$ous sstem

(ils

. "oconut oil6 com*osed mainl o lauric acid 7 is slow to o0idi;e and is resistant to rancidi1cation 4. "anola oil6 low in saturated at 3. (li$e oil6 com*osed mainl o oleic acid and *almitic acid and ot+er att acids . Palm oil6 +as 3< *+enolic com*ounds 7 +as +ig+ saturated $egetable ats 5. Sesame oil6 +ig+ *ro*ortion o *olunsaturated linoleic att acid =. "orn oil6 +ig+ smo-e *oin

/our tests or li*ids . &rease2s*ot test 4. Sa*oni1cation test 3. Acrolein test . >nsaturation test

&rease2s*ot test •





&eneral test or t+e *resence o li*ids Positi$e oil 7 lecit+in 'egati$e water 7 dic+loromet+ane

Sa*oni1cation test •





&eneral test or ester bonds Positi$e oil 7 melted at 'egati$e water

Sa*oni1cation •





?drolsis reaction ?drolsis using a base suc+ as sodium +dro0ide or *otassium +dro0ide Products are . &lcerol 4. [email protected] salts2 salts are soa*s

Sa*oni1able li*ids •





"ontains an ester unctional grou* "an be +drol;ed under basic conditions %ncludes triacglcerols, *+os*+oli*ids, glcoli*ids, s*+ingoli*ids and t+e wa0es

Acrolein test •

&eneral test or ats or glcerin



Positi$e result6 strong odor

>nsaturation test •

&eneral test or t+e *resence o double bonds and t+eir degree o saturation

(becti$es •

to c+aracteri;e li*ids using t+e grease2s*ot test, sa*oni1cation test, acrolein test and unsaturation test.

Met+odolog

Tee, Margarita R. 3PSY5

Grease Spot Test

Tee, Margarita R.

With the use of pencil, four areas of a filter paper was labeled “vegetable oil”, “lecithin”, “H4O” and “dichloromethane” apply a drop of the appropriate substance to the labeled areas

Warm the filter paper

Observations after dropping and after heating

Tee, Margarita R. 3PSY5

Saponification Test

Tee, Margarita R.

! large&si)ed test tubes labeled “oil”, “fat” and “H4O” 8 drops of appropriate substance placed on their corresponding test tube

Add 1 drops of !" #aOH on each test tubes

$lace in boiling water bath for 1%&' mins

$lace in test tube rac( and allow to cool to room temperature Tee, Margarita R.

add %ml of distilled water to each test tube

*topper the test tubes and mi+ vigorously ecord observations after sha(ing whether an emulsion, micelle or clear solution Acidify with few drops of H'*O

Tee, Margarita R.

"i+ with stirring rod #ote the color of the material on top  record the pH of the material, either acidic or basic, using blue and red litmus paper

Tee, Margarita R.

Acrolein Test

Tee, Margarita R.

1 .g of .H*O in a test tube Add % drops of an oil sample or a small piece of solid lipid Heat over a -unsen burner for a few minutes Allow to cool *trong odor

Tee, Margarita R.

Unsaturation Test

Tee, Margarita R.

! large&si)ed test tubes labeled “oil”, “fat” and “glycerol” $lace !ml of /0" on each test tube Add 1 drops of appropriate substance on their corresponding test tube "i+ thoroughly nder the fume hood, add dropwise of %2 bromine&dichloromethane to each test tube while stirring until reddish brown color appears

Tee, Margarita R.

#umber of drops of bromine& dichloromethane and the net number of drops

Tee, Margarita R.

RES>)TS  TA:Y, !ristine "laire :.

&REASE2 SP(T TEST Ater dro**ing GR P

OIL

LECITHI

!C"

H#O

 4 3  5 =  F G <  4

Translucent Translucent Translucent Translucent Translucent Translucent Translucent Translucent Translucent Translucent Translucent Translucent

(*aCue (*aCue (*aCue (*aCue Translucent (*aCue (*aCue (*aCue (*aCue (*aCue (*aCue (*aCue

(*aCueD (*aCueD (*aCueD (*aCue :isa**eared (*aCue (*aCue :isa**eared (*aCue (*aCue (*aCue (*aCueD

Translucent Translucent Translucent Translucent Translucent Translucent Translucent Translucent Translucent Translucent Translucent Translucent

&REASE2 SP(T TEST Ater +eating GR P

OIL

LECITHI

!C"

H#O



Transluce nt Transluce nt Transluce nt Transluce nt Transluce nt Transluce nt Transluce nt

(*aCue

(*aCue

(*aCue

(*aCue

)ig+t stain

:isa**eare d (*aCue

(*aCue 8dried9 (*aCue 8dried9 Hrin-les

4 3  5 = 

Slig+t ellow stain  Translucent

'o stain

(*aCue 8dried9 :isa**eared

(*aCue

(*aCue

Hrin-les

(*aCue

:isa**eare d

:ried

SAP('%/%"AT%(' TEST Ater s+a-ing GR P

OIL

"ELTE! $%T

H#O



Bubbles wit+ ellow inter*+aseI cloud solution Micelle

)ig+t ellow soln

4

Bubbles wit+ w+ite inter*+aseI cloud solution Emulsion

3

Mur- ellow emulsion

"loud ellow micelle



Yellow cloud liCuid

"loud w+ite liCuid

5

Emulsion

Micelle

=  F

Bubbles ormed Emulsion H+ite oam on to*

'o bubbles ormed Micelle H+ite and ellow oam on to*

)ig+t ellow liCuid )ig+t brown soln "olorless Solution "olorless Solution "lear Solution "lear Solution /aint brown soln

SAP('%/%"AT%(' TEST Material on to* GR P

OIL

"ELTE! $%T

H#O



:irt w+ite liCuid H+ite **t Bubbles Bubbles Bubbles, w+ite 1lm )ig+t ellow "loud w+ite H+ite substance H+ite **t :irt w+ite liCuid

 Yellow solids

'one

Yellow **t Yellow s*ots Bubbles ellow laer Bubbles, ellow *owder Yellow Yellow  Yellow ormation

'one 2 2 'one 'one "olorless "olorless

#i$id ellow **t  Yellow solids

'one 'one

4 3  5 =  F G <

SAP('%/%"AT%(' TEST Acid or Base GR P

OIL

"ELTE! $%T

H#O

 4 3  5 =  F G <  4

Acid Acid Acid Acid Acid Acid Acid Acid Acid Acid Acid Acid

Acid Acid Acid Acid Acid Acid Acid Acid Acid Acid Acid Acid

Acid Acid Acid Acid Acid Acid Acid Acid Acid Acid 2 Acid

>'SAT>RAT%(' TEST BrK in :"M GR P

GL&CEROL

OIL

"ELTE! $%T

 4 3  5 =  F G <  4

2 4 45 2  4< 44 = 45 3< F 45

2 55 'SAT>RAT%(' TEST 'et L o dro*s GR P

GL&CEROL

OIL

"ELTE! $%T

 4 3  5 =  F G <  4

4 < < 4< F < 44 < < < < <

F< 3 5 3< 5F < 3  5 
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