Experiment 1: Cell Membrane Diffusion – movement of SOLUTE to higher to lower

concentration
Cell Membrane – lipid-based structure that separates a
cell’s interior from the environment surrounding the cell Osmosis – movement of SOLVENT from an area of higher
concentration to an area of lower concentration and vice
- Also commonly called as the “plasma membrane”
versa
Functions:
Solution:
1. Barrier for keeping the constituents of the cell in
0.1% saline – hypotonic
and unwanted substances out
2. Acts as a gate allowing transport of essential 0.9% saline – isotonic
molecules into the cell and waste products from
25% saline – hypertonic
the cell
Hypotonic – the extracellular fluid has a lower solute
Structure:
concentration (hypo=less than)
1. Membrane lipids which are insoluble in water
- Net flow of solvent will be into the cell
2. ‘head and 2 tails’ structure
3. Lipid bilayer Visible result: swelling of cell
Extracellular – hydrophilic tail Hypertonic – the extracellular fluid has a higher solute
concentration (higher=greater than)
Biomolecules present:
- Solvent will move out from the cell towards region
1. Fatty acid
of higher solute concentration
2. Cholesterol
3. Proteins Visible result: shrinking of cell
4. Oligosaccharides
Isotonic – extracellular fluid has the same solute
Diffusion – water – solvent; KMnO4 – solute (sa concentration to that inside of the cell (iso=same)
experiment)
- No net movement
Result – faster diffusion sa warm water because mas
mataas kinetic energy Visible result: no change in cell size

3 factors: Dialysis – dia(through); lysis(lowering)

1. Temperature – directly proportional sa rate of Dissolution – dialysis

diffusion - Process by which particles are selectively
2. Molecule size and shape – inversely proportional removed from a liquid by consequence of their
bc larger particles moves slower leading to lower differing capacity to pass through a membrane into
rate of diffusion another liquid
3. Magnitude of concentration diffusion – molecules
from higher concentration moves to lower Dialysate – water outside the semipermeable membrane
concentration Dialyser or dialysing membrane – filter (cellophane)
Higher concentration difference – faster movement and - Protein and sugar are not capable of passing
higher diffusion rate through the dialyser due to their large size
Lower concentration difference – slower movement and (macromolecules)
lower diffusion rate (directly proportional) 1. Heller’s Test (proteins) negative
o Negative result: absence of white ring
Equilibrium = equal o Positive: formation of white ring at the
junction
HNO3 – denature the protein Galactose – monosaccharide that forms the disaccharide
lactose
A white ring appears if protein is present
- Patients with galactosemia cannot metabolize this
2. Fehling’s test (sugar) – negative sugar
o Negative result: no red ppt - Buo parin yung carbohydrate na lalabas pag
o Positive result: brick red ppt kumain sila (lactose intolerance)
- Sucrose – non reducing sugar - In severe cases, it can be abnormally stored in
- Reagent: Fehling’s A – copper (II) sulfate (blue) pancreas and liver which can cause liver cirrhosis
Fehling’s B – Rochelle salt made in (scarring)
NaOH - Can also form cataracts; cornea will be filled with
o General test for monosaccharides and blood sugar; bumabagal kase blood flow mo
other reducing sugars - Commonly associated with diabetes
- Can detect galactosemia through newborn
3. Barium Chloride Test (salts) – positive
screening
o Negative: no white ppt
o Positive: white ppt (or) turbidity To prevent galactosemia – physician will supplement foods
- Sodium sulfate was able to pass through the without sugars that you cannot metabolize
membrane (galactose/lactose dietary sources)
- Barium chloride will react against sodium sulfate Frucose – fruit sugar
forming a white ppt (barium sulfate)
- Barium chloride + sodium sulfate = sodium - Found in honey and is twice as sweet as normal
chloride + barium sulfate (white ppt) table sugar
- Can be utilized sperm cells for energy which
Experiment 2: Monosaccharides and Disaccharides provides motility; Less semen fructose is less
motility; often leads to infertility
Carbohydrates – commonly referred to as sugars and 2. Disaccharides – carbohydrates composed of 2
starches monosaccharides
- Are polyhydroxy aldehydes and ketones, or o Maltose – glucose and glucose
compounds that can be hydrolyzed to them o Lactose – glucose and galactose
- Cn(H2O)n – hydrates of carbon o Sucrose – glucose and fructose

Classification of Carbohydrates Maltose – gets its name from malt, the liquid obtained from
barley used in the brewing of beer
1. Monosaccharides – simplest carbohydrates which
contains only a single aldehyde or ketone - Produced from the hydrolysis of starch
functional group. Lactose – found in milk from both humans and cows
o Glucose, galactose, fructose
o Also classified according to number of - Not relatively sweet
carbon in its chain - Lactose intolerance – patients lacking the enzyme
o # of carbon – triose, tetrose, pentose, “Lactase” cannot metabolize the sugar properly
hexose
Sucrose – also known as cane sugar
Glucose – also called as dextrose
- Most common disaccharide found in nature
- Component of polysaccharides cellulose, starch, - Derivatives of sucrose are present in artificial
and glycogen as its building block sweeteners
- In medicine, glucose is called blood sugar - Examples of these derivatives include sucralose,
- Normal: (70-110 mg/dL) aspartame, saccharin
- Primary source of energy for living cells
- Glycogen (storage form of glucose); pancreas and General laboratory tests
livers 1. Molisch’s Test – test detects the presence of
carbohydrates in a given solution
Reagent: - Aldehyde carbonyl groups form new C=O bonds
through addition of oxygen atoms by oxidizing
 Alpha Naphthol dissolved in ethanol (main agents
reagent) purpose: color indicator - Carbohydrates reduce the oxidizing agents
 Concentrated HSO4 – dehydrating agent - Aldoses are oxidized into aldonic acids
Glucose, Lactose, Sucrose – purple ring at the junction of - Aldehydes oxidized under alkaline conditions into
2 liquids; so they are all positive for carbohydrates a carboxylate anion

2. Schiff’s Test – to detect the presence of aldehyde Tests for reducing property
functional group 1. Tollen’s Test
Reagent: Para-rosanilin with hydrochloride o Principle: Oxidation by silver ion
o Reagent: Silver Nitrate in Ammonia Water
Positive result: purple/magenta colored solution (ammoniacal AgNO3) as an oxidizing
agent
Glucose and Lactose: purple solution (positive)
o Positive Result: Formation of Silver Mirror
Sucrose: no purple solution (negative)
Glucose + Tollen’s reagent – gluconic acid + ammonium
Oxidation Reduction reactions Test nitrate
LEORA GEROA or LEO says GER Silver ions lose its positive charge so it becomes silver
metal
Loss of electrons is oxidation – gain of oxygen; from ion to
metal Carbohydrate reduced the tollen’s reagent
Gain of electrons is reduction – loss of oxygen; from metal
to ion
2. Fehling’s test – oxidation by copper ion
Oxidation – aldehyde carbonyl groups present in Reagents: Fehling’s A and Fehling’s B
carbohydrates are oxidized to form a carbonyl group Fehling’s A: Hydrated CuSO4 solution
Fehling’s B: Potassium sodium tetrahydrate or
- Increase in C=O bonds or decrease in C-H bonds Rochelle salt in NaOH solution
- RCHO to COOH
- Fehling’s and Benedict’s Test Glucose reduced the Fehling’s reagent; Fehling’s reagent
converts glucose into gluconic acid
Reduction – aldehyde carbonyl groups present in
carbohydrates are reduced to primary alcohols using H2 in 3. Benedict’s Test
the presence of palladium metal (Pd)
Reagents:
This alcohol is termed as “alditol” or sugar alcohol
Anhydrous sodium carbonate (complexes with Cu+2 during
- Decrease in C=O bonds, increase in C-H bonds storage to avoid deterioration)
Reducing Agents – compounds that causes the loss of Sodium citrate (stabilizer or buffer)
oxygen from an oxidizing agent
Copper (II) sulfate pentahydrate (source of copper ion)
- Oxygen is gained by the reducing agent
Clinitest – tablet test then mix with urine, andun na lahat
- CuSO4 – oxidizing agent
ng components ng reagent
- CuSO4 is reduced in the process by glucose by
taking the oxygen Negative result: blue solution
Oxidizing agents – oxygen is lost Positive: brick red precipitate
- Compounds that causes the gain of oxygen to a - Detection of reducing sugars in a pathologic urine
reducing agent samples
- Commonly observed in diabetes mellitus type 2
Carbohydrates as a reducing agent
(glucose in urine)
High Blood Glucose Kidneys – nephrons (basic functional Antigens are also called as agglutinogen
units)
Antibodies – agglutinins
160-180 mg/dL – glucose renal threshold
Sugar markers – 2 molecules of D-galactose, L-fucose, N-
- Pag nagexceed sa limit dun na madedetect sugar acetyl-D-glucosamine
sa urine
4. Mooore’s Test – siya lang may odor sa results Type A – N-acetyl-D-galactosamine
- Carbohydrates react with alkaline reagents like
Type B – D-galactose
NaOH
- Oxidation of glucose into gluconolactone The ABO blood type of each person is determined by a
- Chemical caramelization of sugars single gene
- Gluconolactone is found in cosmetics and food
additives For type A, there is a gene for GTA, a glycosyltransferase
- Positive result: yellow/orange/golden brown that adds N-acetyl-D-galactosamine
solution with caramel odor
5. Picric Acid Test For type B, there is a gene for GTB, a different
glycosyltransferase that adds galactose.
Reagents include:
Type O – neither enzyme is made
 Picric Acid
 NaCO3 (increases pH) The A gene codes for a protein that adds N-acetyl-D-
 Picric Acid (yellow) is reduced into picramic acid galactosamine in the a1,3 glycosidic bond to the N-acetyl-
D-galactosamine residue at the end of H antigen.
Positive result: mahogany red
The B gene codes for a protein that adds galactose in the
a1,3 glycosidic bond to the galactose residue at the end of
6. Nylander’s Test H antigen.
Reagent: Since both A and B genes are present in AB individuals,
some H antigens get a N-acetyl-D-galactosamine residue
 Bi(OH)2NO3 or Bismuth subnitrate
while others get a galactose residue
 potassium sodium tartrate
 KOH O have neither A nor B gene so no additional carbohydrate
- Sugars reduced Bi(OH)3 from Bismuth subnitrate is added to the H antigen
into black metallic bismuth on heating
H antigen – precursor
Positive result: Metallic Bismuth black precipitate
AB – universal recipient because wala siya antibodies
Experiment 3: Monosaccharides in blood groups
O – universal donor cause wala siya antigen
Monosaccharides – building blocks for polysaccharides
Type A – Antigen A and antibodies B
- Functions as receptors and markers
- Also serves as biochemical bases for various Type B – Antigen B and Antibodies A
blood groups on the surface of RBC’s
AB – Antigen A and B, no antibodies
Agglutination – when antibodies (proteins) in the plasma
O – no antigens, antibodies A and B
are mixed with their corresponding antigens on the surface
of large, easily sedimented particles such as animal cells, Major – PRDS (patient’s red cells and donor’s serum)
erythrocytes or bacteria the antibodies cross link the
Minor – PSDR
paticles forming visible clumps.
RBC’s – antigen
Plasma – antibodies