Basic Lab Operation
Separation of solids from liquids
A Centrigugation :
Centrifuge – principle- The working of centrifuge is based on the
principle of centrifugal force, which act on a substance in circular
motion, towards the periphery.
The factors which govern the speed of centrifugation are
1. The revolution per minute.
2. Length of Radius- the radius is measured from center of rotation to
the inside bottom of tube.
3. Shape and size of particles
Separation of solids from liquids
• 4. viscosity and specific gravity of fluid under centrifugation.
• 5. Gravitational force acting on the particles.
Uses :- The centrifuge is mainly used for
1. Separation of serum or plasma from red blood cells.
2. Separation of sediment in urine.
3. Separation of protein free filtrate.
4. Washing of red blood cells by normal saline.
Separation of solids from liquids
• Different type of centrifuges –
1. The swing out head type (Horizontal rotar)
2. Angle head type ( fix angle rotor )
A rotor is the device used to hold centrifuge cups and tubes.
The swing out head type (Horizontal rotar): Tubes are in vertical
position when the centrifuge is at rest and assumed a horizontal
position when the centrifuge is at rest. The speed > 5000 RPM is
generally not practical in swing head centrifuge due to excess heat
developed by friction of tubes with air.
Angle head centrifuge : tubes are positioned rigidly at a fix angle to
the vertical axis of rotation. Due to the very low friction of air, angle
head rotor units can attain much higher speed.
• -The usual expression of RPM (Revolution per minute) gives only the
centrifugation speed and does not express the centrifugal force. A
better expression of forces generated by centrifuge is RCF (relative
centrifugal force)which calculated by considering the speed of
rotation (RPM) and the radius from the center of rotation (R ).
• RCF =R X (RPM)2 X 118 X 10 -7+
CARE AND MAINTANANCE
1. Place the centrifuge on firm base. It should not be placed near a
sensitive equipment such as photometer and
spectrophotometerbecause it generate electrical noise and high
current drain, when started up. It should not be run near
combustible fluid.
2. Before the centrifugation, the centrifugation tubes and the cups
should be balance properly. Do not run the centrifuge with buckets
or tubes missing from the units.
3. The chamber should be kept clean. All the spills should be cleaned
immediately.
CARE AND MAINTANANCE
• 4. Always make sure that the lid is closed while the centrifuge is
operating. Never open lid until the rotor has come to a complete stop.
• 5. Observe for usual normal vibration noise during operation. The
centrifuge may vibrate excessively, if the tubes inside are not
balanced properly. This should check immediately.
• Keep tachometer to measure RPM of the machine.
Separation of solids from liquids
• B. Filtration : is the process separating solid particals from the liqude
in which the are suspended. In this process the suspension is poured
on a porous material called filter. The liquid passes through (called
filtrate) the filter retains the solid particales ( called residue) filer
paper is most commonly used for [Link] id folded and placed on a
funnel
• Funnels : these are available in verity of the separation of
• Solid from liquids
Weighing
• Balance : a balance is an important instrument in the laboratory
which measures the weight of a substance.
• Balances are of different sensitivities depending on there use.
• The physical balance is less sensitive than the analytical balance.
• For routine lab purposes, the sensitivity of a balance can be consider
to be the smallest mass that can be weight accurately.
BALANCES
• Type of physical balances balances.
1. Two pan balance
2. Mono-pan balance
BALANCES
• 1. Physical balances – (open two pan type) are used for weighing out
substances, where great accuracy is not required. These are useful for
the preparation of qualitative reagents.
• . Two pan analytical balances are useful for the preparation of
quantitative reagents. Weighing range from 0.001 to 100 gms can be
obtained on this balance.
• The mono-pan balances are most sensitive and weighing accuracy
can be obtained up to 0.001 mg.
BALANCES
• Analytical balance These are mainly used to weight chemicals
accurately for the preparation of analytical reagent.
• It is an instrument for determining mass of a body by equilibrium of
weights suspended from the opposite sides of a bar having fulcrum at
its center and two arm of equal length.
• Electrical balances. It consist of 3 basic component systems a. a null
detector b. a feedback loop to control the balancing force and a read
out device.