Characterization of Saponiable 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
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%ntroduction RE&%'(, Paul Andrew R. 3PSY5
)i*ids •
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)i*ids are naturall organic com*ounds t+at are insoluble in water but soluble in organic sol$ents. Animals li*ids ma-e u* most o t+e ats in t+e orm o adi*ose tissues w+ic+ +el*s in insulation and t+ermal regulation. /ats are -nown to store +eat. Plants li*ids are seen as t+e wa0
"lassi1cation o li*ids •
(*en2c+ain com*ounds wit+ *olar +eads and long non*olar tails . 4. 3. . 5.
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/att acids Triacglcerols S*+ingoli*ids P+os*+oli*ids &lcoli*ids
/used2ring com*ounds . Steroids
"lassi1cation o li*ids •
(n t+e basis o w+et+er t+e will undergo +drolsis in al-aline solution 1. Saponiable lipids
4. 'onsa*oni1able li*ids
/att Acids Polar end6 t+e carbo0l grou* 'on*olar tail6 t+e +drocarbon c+ain /att acids are am*+i*at+ic T+e usuall +a$e e$en number o carbon atoms 7 t+e +drocarbon c+ain is unbranc+ed
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Two main t*es Saturated 7 unsaturated
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Triaclglcerols T+e 3 alco+ol grou*s in glcerol 8glcerol contains 3 +dro0l grou*s9 orms ester lin-ages wit+ att acids. /unction storage 8es*eciall in animals9
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P+os*+oli*ids T*e o li*id t+at contains a *+os*+orus atom attac+ed to t+e glcerol bac-bone T+e are -nown to be t+e main li*id com*onent o t+e cell membrane 4 t*es o *+os*+oli*ids
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. P+os*+oaclglcerols2 glcerol is t+e bac-bone 4. S*+ingomelins2 s*+ingosine instead o glcerol
&lcoli*ids •
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A li*id to w+ic+ a sugar moiet is bonded A carbo+drate is bound to an alco+ol grou* o a li*id b a glcosidic lin-age "eramides are t+e *arent com*ounds or glcoli*ids
S*+ingoli*ids •
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:o not contain glcerol but instead t+e contain sphingosine. /ound bot+ in *lants and animals 7 t+e are abundant in t+e ner$ous sstem
(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 $egetable ats 5. Sesame oil6 +ig+ *ro*ortion o *olunsaturated linoleic att acid =. "orn oil6 +ig+ smo-e *oin
/our tests or li*ids . &rease2s*ot test 4. Sa*oni1cation test 3. Acrolein test . >nsaturation test
&rease2s*ot test •
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&eneral test or t+e *resence o li*ids Positi$e oil 7 lecit+in 'egati$e water 7 dic+loromet+ane
Sa*oni1cation test •
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&eneral test or ester bonds Positi$e oil 7 melted at 'egati$e water
Sa*oni1cation •
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?drolsis reaction ?drolsis using a base suc+ as sodium +dro0ide or *otassium +dro0ide Products are . &lcerol 4. Sodium@Potassium salts2 salts are soa*s
Sa*oni1able li*ids •
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"ontains an ester unctional grou* "an be +drol;ed under basic conditions %ncludes triacglcerols, *+os*+oli*ids, glcoli*ids, s*+ingoli*ids and t+e wa0es
Acrolein test •
&eneral test or ats or glcerin
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Positi$e result6 strong odor
>nsaturation test •
&eneral test or t+e *resence o double bonds and t+eir degree o saturation
(becti$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 Ater 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 Ater +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 Ater s+a-ing GR P
OIL
"ELTE! $%T
H#O
Bubbles wit+ ellow inter*+aseI cloud solution Micelle
)ig+t ellow soln
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Bubbles wit+ w+ite inter*+aseI cloud solution Emulsion
3
Mur- ellow emulsion
"loud ellow micelle
Yellow cloud liCuid
"loud w+ite liCuid
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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 soln "olorless Solution "olorless Solution "lear Solution "lear Solution /aint brown soln
SAP('%/%"AT%(' TEST Material on to* GR P
OIL
"ELTE! $%T
H#O
:irt w+ite liCuid H+ite **t Bubbles Bubbles Bubbles, w+ite 1lm )ig+t ellow "loud w+ite H+ite substance H+ite **t :irt w+ite liCuid
Yellow solids
'one
Yellow **t Yellow s*ots Bubbles ellow laer Bubbles, ellow *owder Yellow Yellow Yellow ormation
'one 2 2 'one 'one "olorless "olorless
#i$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 << 2 == 35 G5 4< 4< 3< 5F 4FG
2 2 2 4= =< 4 =< F 4G 5< 44
>'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 << 5<
45 2 = F < 5 2 42 23
:iscussion REP(RTER Terrence Julian B. Pestao
PESTAN(,
&rease O S*ot Test A sim*le test in determining t+e *resence o li*ids Rea'ents( :"M, ?4(, )ecit+in, (il sam*le
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Principle( #a*ori;ation o $olatile
substances )isible Res*lt( Translucent Stains Positi+e res*lts in( (il Sam*le
and )ecit+in
&rease O S*ot Test •
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#olatilit >nsaturation 'umber o "arbons
PESTAN(,
Triglceride
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)E"%T?%'
Sa*oni1cation Test •
%n$ol$es saponifcation O +drolsis using a strong base
Lipids (at, oil)
Glycerol and Soap PESTAN(,
Sa*oni1cation Test
PESTAN(,
PESTAN(,
Sa*oni1cation Test Rea'ents( 'a(? 8strong
base9, ?4S( Principle( base +drolsis )isible Res*lt( /ormation o
bubbles @ *reci*itate Positi+e res*lts in( Melted
/at and (il PESTAN(,
Saponication *,ber- )al*e a./.a. 0oettstorfers *,ber2 •
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O is t+e needed mg o 'a(? 8or !(?9 to neutrali;e t+e att acids rom t+e +drol;ed g o t+e li*id "an gi$e notions regarding t+e structure o att acids •
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Molecular weig+t "+ain )engt+
The hi'her the saponication n*,ber 324 the lo5er the ,olec*lar 5ei'ht 62
8more att acids are reed *er gram o at ater +drolsis9 PESTAN(,
PESTAN(,
Reerences Retri$ed+tt*@@www.orinag.c+@inde0.*+*Q sid6(#5cSt&'Eb!)s)3baR5=>0g55C$P(lReb7c5*6G 57c5l6en. 8'o$ember 4=, 4<59. "am*bell, M.!. 7 /arrell, S.(. 84<4@. Bioc+emistr. 8t+ ed.9. "aliornia, >SA Broo-s@"ole, "engage )earning.
Concl*sion )illan*e+a4 0er+in 7.
Ob8ecti+es To
characterize a fat or an oil sa,ple *sin' the 'rease9spot test4 saponication test4 acrolein test4 and *nsat*ration test.
The
'ro*ps 5ere able to characterize a fat or an oil sa,ple *sin' the 'rease9spot test4 saponication test4 acrolein test4 and *nsat*ration test.
The
'rease9spot test as a 'eneral test for the presence of lipids4 5o*ld 'i+e a positi+e res*lt in both the oil sa,ple and lecithin b*t a ne'ati+e res*lt in 5ater and !C"!ichloro,ethane2.
The
saponication test as a test for the presence of ester bonds4 5o*ld 'i+e a positi+e res*lt in both the oil sa,ple and ,elted fat b*t a ne'ati+e res*lt in 5ater.
The
*nsat*ration test as a test for the presence of do*ble bonds and the de'ree of sat*ration4 5o*ld ha+e di:erent n*,ber of drops of bro,ine9 dicholoro,ethane in *nsat*ratin' the 'l;cerol4 oil and ,elted fat *ntil the; colorize to a red9bro5n sol*tion.
The
acrolein test as a test for the presence of 'l;cerol4 5o*ld 'i+e a positi+e res*lt in lecithin.
So*rces of Error inaccurate measurement o t+e reagents used obser$ational error
