Bi0chEm LAB - Lipids

polymers

LIPIDS is water insoluble biomolecule associated into __

hydrophobic or amphipathic

tend to form surface monolayer, bilayer, micelles, or vesicles, when in contact with water.

Phospholipid bilayers

Glycolipids

Cholesterol

LIPIDS Consist of

Glycolipids

Lipid connected to a glyco- group or carbohydrate molecule

Cholesterol

Provides fluidity to the membrane.

Glycolipids

Provides stability to the membrane

Phospholipid bilayers

polar heads and nonpolar tail

Phospholipid bilayers

Lipid connected to a phosphate group

50%

how many % of our cell membrane is composed of lipids

LIPIDS

It serves as our structural framework of cell membranes/plasma membranes.

true

tof. lipids are oils and very important to our diet (moderation)

Castor oil (Cosmetic & excipient)

● Good hair grower

● Use as an pharmaceutical excipient

Cod Liver Oil (Emulsion)

● Emulsion

Total Parenteral Nutrition (TPN)

For people who are not able to take

food as source of their energy

For compounding semisolid dosage

forms

Anhydrous lanolin (as a excipient) is for

For compounding semi-solid products

(cream, ointments)

Beeswax (Common excipients) is for __

true

tof. Steroids are considered lipids

Fluticasone Propionate

Nasal Corticosteroids - steroid example

thermal insulator

Lipids serve as a thermal insulator in our body. More fat(lipids) in the body, 'di kaagad giniginaw

lipids

If the body has insufficient carbs intake, the body will use __ as an alternative

low-carb

LIPIDS most undergo a __ diet if they want to lose weight.

INSULATION

ENERGY STORAGE

STRUCTURAL COMPONENT OF CELL MEMBRANE

ACT AS SIGNALING MOLECULES (stero, arachi, leptin)

Give functions of lipids

Arachidonate

responsible for producing prostaglandin (signaling molecule)

Leptin

endogenous molecule in the body; regulates several body functions (blood pressure, menstrual cycle)

PROTECTION

INSULATION

ENERGY STORAGE

SIGNALING MOLECULE

REMEMBER BASIC FUNCTION OF LIPIDS: P I E S

FATTY ACIDS/FATTY ACYLS

Basic units of lipids

Linear

FATTY ACIDS/FATTY ACYLS is a __ carboxylic acid molecules

Triacylglycerols

Glycerophospholipids

Sphingolipids

Glycoglycerolipids

Waxes

SAPONIFIABLE - With Fatty acids

Eicosanoids

Steroids

Lipid-soluble vitamins (Vit. A,D,E,K)

Terpenes (extracted from plants)

NONSAPONIFIABLE - Without Fatty acids

These are signaling molecules;

vit. ADEK

Lipid-soluble vitamins

saturated

lipids that comes from fats of animals

& also plants; SOLID in nature

UNSATURATED

Fixed oils extracted from plants; LIQUID in nature

false. it is

Soluble in non-polar organic solvent (chloroform, ether, benzene)

tof. FATTY ACIDS is insoluble in ether

increases

As the number of Carbon increases, solubility in organic solvent __ (more lipophilic)

increase

As the number of Carbon increases, MP & BP __ (C-C), more lipophilic.

false. Presence of unsaturation decreases MP

tof. Presence of unsaturation increases melting point

UNSATURATED fatty acyls

Compact; exist as solid - SATURATED

Not compact; exist as liquids

ESTER FORMATION

HALOGENATION

HYDROGENATION

OXIDATION

REACTION OF FATTY ACIDS

Acyls (carboxylic acids)

subjected to Ester formation

true

tof. trans fat increases cardiovascular risk.

Rancidification

oxidation reaction produces what?

Rancidification (OXIDATION reaction with Kmno4- mild oxid)

slow & spontaneous reaction of double bonds with atmospheric O2 or w/ certain oxygenases to form short chain aldehydes responsible for objectionable odor & unpleasant taste

aldehydes & carboxylic acids.

Ozonolysis is a reaction for double bond w ozone to form __ and __

Chloroform: methanol mixture (2:1)

Anhydrous Na2SO4

ISOLATION AND EXTRACTION

REAGENTS:

Anhydrous Na2SO4

used to remove residual water (desiccant)

Chloroform: methanol mixture (2:1)

Extracting solvent

oxidation & peroxidation

In isolation of lipids, it is susceptible to __ and __

1. penetrate the tissue

2. break protein-lipid bond.

In extraction, SOLVENTS to be used must :

- Homogenization by triturating with sand

-or using an osterizer/blender

breaks the integrity of the cell's membranes

true

TOF. Non-lipid components can be removed by SIMPLE WASHING while lipids remain DISSOLVED in the organic solvent

true

tof. Non-lipids are most likely water soluble, thus can be removed by simple washing

below 4 degrees

Tissues can be cooled to below __ degrees immediately following excision

differences in solubility.

lipids are separated based on their __

Lipids are water-insoluble but SOLUBLE in ORGANIC solvents.

tof. Lipids are water-insoluble and insoluble in organic solvents.

TRIACYLGLYCEROLS

PHOSPHOGLYCERIDES

GLYCOLIPIDS

CHOLESTEROL

SOLUBILITY OF LIPIDS IN ORGANIC SOLVENTS which are:

extracted w/acetone or diethyl ether

TRIACYLGLYCEROLS is extracted with

Lecithins

PHOSPHOGLYCERIDES is also called

-insoluble in acetone

-soluble in ether, chloroform, benzene, hot alcohol, carbon disulfide

PHOSPHOGLYCERIDES is

insoluble in ___

soluble in ___

insoluble in ETHYL & METHYL ALCOHOLS

Cephalins has same solubility as lecithins but are insoluble in _ _AND __

ether, benzene, chloroform * hot alcohol and easily crystallize from such solutions

CHOLESTEROL is soluble in

true. Triglycerides are long-term energy storage molecules because they will not mix with water and break down.

tof. Triacyglyceros cannot be extracted while the remaining can be.

false. they do not USUALLY. kaya we dont use acid and enztme hydrolysis

tof. lipids undergo hydrolysis.

Requires two solvent system

TWO DIMENSIONAL TLC has how many system

90° clockwise

SECOND DEVELOPMENT

After letting the used tlc dry (yung may spot na) rotate it __

IODINE VAPOR

NINHYDRIN SPRAY

VISUALIZING AGENTS

IODINE VAPOR

It detected yellow-brown spots which also detects unsaturated lipids. What visualizing agent is used?

Purple spots

TLC results to purple spots

- Lipids containing free amino groups

○ Phosphotidylethanolamine (PE)

○ Phosphotidylserine (PS)

what visualizing agent is used?

iodine vapor chamber (REMEMBER!!)

(Beaker with iodine crystals covered by watch glass)

alternative for uv light in visualizin

Ninhydrin

commonly used in the analysis of amino acids and peptides, specifically for their detection and visualization.

COLUMN CHROMATOGRAPHY OF LIPIDS

Used to separate the different components of the extract

1.) Petroleum ether: Ethyl ether

2.) 5% Methanol in DCM

3.) DCM: CH3OH: H2O (1:3:1)

By the end of the Column Chromatography, you will have 3 eluate:

Qualitative Analysis

1.) Petroleum ether: Ethyl ether

2.) 5% Methanol in DCM

3.) DCM: CH3OH: H2O (1:3:1)

○ These 3 eluates will undergo __

Glycerophospholipids

Sphingolipids

Eluent : DCM : Methanol : Water (1:3:1)

for?

Cholesterol

Eluent: 5% Methanol in DCM

for?

Triacylglycerides (TAGs) ACROLEIN TEST

eluent: Petroleum Ether: Ethyl Ether (9:1)

for?

Ferric Hydroxamate Test

TEST FOR ESTER is also called

Ethanol:1-butanol (3:1) NH2OH ·HCl; NaOH 5% FeCl3 ·6H2O

TEST FOR ESTER

REAGENT

Burgundy color of ferric hydroxamate complex

VISUAL POSITIVE RESULT

hydroxamic acid

For Ester, upon exposure to Hydroxylamine, will be converted to __

Ferric Chloride,

Then once Hydroxamic acid is exposed with __, this will form a complex which we can see as Burgundy in color (+).

Ester's exposure to Hydroxylamine, converted to hydroxamic acid. Hydroxamic acid is exposed with Ferric Chloride = burgundy

causes burgundy color

ACROLEIN test

TEST FOR GLYCEROL also called

KHSO4 (Potassium Bisulfate)

100°C water bath

TEST FOR GLYCEROL/acrolein REAGENTS

Triacylglycerides (TAGs)

Glycerophospholipids

TEST FOR GLYCEROL/acrolein

is test for?

DEHRYDRATION

● Nawalan ka ng hydrogen and nag oxidize

OXIDATION

TEST FOR GLYCEROL/acrolein

principle:

TEST FOR GLYCEROL/ACROLEIN

TEST with VISUAL POSITIVE RESULT of:

Burnt fat/Acrid/Irritating odor

KRAUT'S TEST

TEST FOR CHOLINE aslo called

Bismuth nitrate

HNO3 (nitric acid)

Kl (potassium iodide)

TEST FOR CHOLINE/Krauts REAGENTS

Choline in Phosphadyl choline Sphingolipids

ST FOR CHOLINE/Krauts test for?

1. Proposed condensation, Dehydration and epimerization

2. Modification of Dragendorff's test, which is a test for alkaloids

TEST FOR CHOLINE KRAUT'S TEST

principle

TEST FOR CHOLINE/KRAUT'S TEST

VISUAL POSITIVE RESULT of Orange precipitate. what test?

Vitamin C or ASCORBIC ACID

TEST FOR PHOSPHATE (ASCORBIC ACID) also called

HNO3

NaOH

Molybdate reagent

Ascorbic Acid Solution

TEST FOR PHOSPHATE ASCORBIC ACID reagents

Phospholipids

TEST FOR PHOSPHATE ASCORBIC ACID is a test for __

orthosphospate

Ammonium Molybdate, Antimony Potassium Tartrate→ They react in acid medium with __ to form a heteropoly acid-phosphomolybdic acid- that is reduced to intensely colored molybdenum blue by ascorbic acid

LIEBERMANN BURCHARD TEST

TEST FOR CHOLESTEROL also called

Concentrated H2SO4

Acetic anhydride

TEST FOR CHOLESTEROL/LIEBERMANN BURCHARD TEST reagents:

Cholesterol

Cholesteryl esters

TEST FOR CHOLESTEROL/LIEBERMANN BURCHARD test for?

Complexation reaction

principle of TEST FOR CHOLESTEROL LIEBERMANN BURCHARD TEST

Emerald green Solution

VISUAL POSITIVE RESULT of CHOLESTEROL LIEBERMANN BURCHARD TEST