EXPT 4B

HYDROLYSIS OF NUCLEIC ACID

Luzviminda Gonzales-Ranay
 HYDROLYSIS OF DNA MOLECULE
• Acid Hydrolysis (Lysis – break down)
– Depurination
• break down of glycosidic bonds only if pH > 3
• Products are: purine base and deoxyribose with
phosphate group

– Total breakdown
• break down of glycosidic bond with phosphodiester
bond if pH < 2
• Extremely low pH hydrolyses DNA completely
• Products are: phosphate group, the purine or
pyrimidine base, and deoxyribose
 QUALITATIVE ANALYSIS
• Benedict’s Test
– General test for reducing
sugars
– Reagents: CuSO4, Na2CO3 and
sodium citrate
– Sugars are oxidized while
Cu2+ is reduced to Cu +
forming Cu 2O precipitate
– Green to orange (depending
on the amount)
– Test for presence of
deoxyribose
 QUALITATIVE ANALYSIS
• Test for Purine Bases
– Test for the presence of Adenine and Guanine
– Reagents: Silver Nitrate and Ammonium
Hydroxide
– Silver ion in ammoniacal solution precipitates
purine bases reacting to the nitrogen of purine
base
 QUALITATIVE ANALYSIS
• Bial’s Test
– Test for pentoses
– Reagents: Orcinol,
hydrochloric acid,
FeCl3
– Blue or green
signifies positive
result
 QUALITATIVE ANALYSIS
• Test for Phosphate
– Reagents: Ammonium
hydroxide, nitric acid,
ammonium molybdate
– Yellow precipitate is the
positive indicator

PO4 -3 + 12(NH4 ) 2 MoO4 + 21HNO 3 (NH 4 ) 3 PMo12O40•6H2 O + 21NH 4 NO3 + 6H 2 O

(yellow precipitate)
 EXPT 5A
LIPIDS
Luzviminda Gonzales-Ranay
 LIPIDS
Substances from living organism that are
soluble in non-polar solvents (ether, chloroform,
benzene, & acetone) but not in water.
 BIOLOGICAL FUNCTIONS OF LIPIDS
1. Lipids in form of a bilayer are essential components of
biological membranes.
a. Structural components of cell membranes
b. Protective functions in bacteria, plants, insects, & vertebrates,
serving as a part of the outer coating between the body of the
organism and the environment.
2. Lipids containing hydrocarbon side chains serve as
energy stores
3. Many intra- and intercellular signaling events involve lipid
molecules
a. Regulatory substances
b. Transport form of neurotransmitters
c. Determinants of immunological specificity
d. Enzyme cofactors
e. Receptors in nerve ending membranes
f. Chemical messengers.
 CLASSES OF LIPIDS
I. SIMPLE
A. Triglycerides (Fats and
Oils)
– esters of fatty acids and
glycerol
B. Waxes
– esters of fatty acids and
monohydric alcohol

Cetyl Palmitate
 CLASSES OF LIPIDS
II. COMPOUND
A. Phospholipids
– esters of fatty acids with
glycerol containing an
esterified phosphoric acid and
a nitrogen
B. Glycolipids
- contain an amino alcohol
(sphingosine or iso-
sphingosine) attached with an
amide linkage to a fatty acid
and glycosidically to a
carbohydrate (sugars, amino
sugar, sialic acid)
 CLASSES OF LIPIDS
III. DERIVED
- Substances derived from
hydrolysis of simple and
compound lipids
A. Fatty Acids
B. Steroids
C. Terpenes
D. Carotenoids
 SAMPLES
• Olive oil – predominantly composed of oleic acid

• Coconut oil – predominantly composed of lauric acid

 SAMPLES
• Lecithin - Phosphoglycerides that contains the aminoalcohol

• Butter – fatty acids composition are: myristic, palmitic, and

oleic
 SAMPLES
• LOTION
– Mineral Oil
– Glycerol
– Water
– Additives
 TRANSLUCENT SPOT TEST
A translucent spot test is also a
preliminary test for the lipids
which can be detected by the
appearance of a translucent and
greasy spot. Principle: The lipid
will not wet the filter paper, unlike
water. The lipid will form a greasy
spot as they are having a greasy
texture that will penetrate into the
 SUMMARY

SAMPLE RESULTS

Olive Oil Positive

Coconut Oil Positive

Lecithin Positive

Butter Positive

Lotion Negative
 SOLUBILITY
Principle: Solubility test is based on the property of
lipid to dissolve in different solvents. Lipids are readily
miscible in non-polar solvents like chloroform, partially
soluble in a polar solvent like ethanol and immiscible
in a polar solvent like water.

Positive result: Lipids are soluble in

non-polar solvent i.e. chloroform and
partially soluble in ethanol which can
solubilize upon heating.

Negative result: Lipids are insoluble in

a polar solvent i.e. water.
 SUMMARY

Samples Water Dichloro- Cyclo- Dilute HCl Dilute

(Polar) methane hexane (Ionic) NaOH
(Slightly (Nonpolar) (Ionic)
Polar)

Olive Oil Insoluble Soluble Soluble Insoluble Insoluble

Coconut Oil Insoluble Soluble Soluble Insoluble Insoluble

Lecithin Insoluble Soluble Soluble Insoluble Insoluble

Butter Insoluble Soluble Soluble Insoluble Insoluble

Lotion Soluble Insoluble Insoluble Soluble Insoluble

 UNSATURATION TEST
• Unsaturation test is used to detect the
presence of unsaturated fatty acids or the
amount of double bond in a lipid sample.
 SUMMARY

SAMPLE RESULTS

Olive Oil Positive

Coconut Oil Negative

Lecithin Negative

Butter Positive

Lotion Positive
 ACROLEIN TEST
• Detects the presence of glycerol or fat.
• For positive result, addition of lipid sample
potassium bisulfate (KHSO4), the glycerol portion of
the molecule is dehydrated to form an unsaturated
aldehyde, acrolein that has a pungent irritating
odor.
 SUMMARY

SAMPLE RESULTS

Olive Oil Positive

Coconut Oil Positive

Lecithin Positive

Butter Positive

Lotion Positive