PHARMACEUTICAL ENGINEERING CENTRIFUGATION

#centrifugation centrifuge #pharmaceutical #pharmaceuticaltechnology #pharmaceuticalengineering #pharmacy #bpharmacy #bpharm #ultracentrifuge #perforatedbasket #nonperforatedbasket #pharma_e_notes #engg #p'engg #supercentrifugation

1. CENTRIFUGATION. 9150138755 SUBMITTED TO:MR.L.GOPI BY: N.PRADHIKSHA B.PHARMACY SECOND YEAR(SEM-III).

2. SYNOPSIS ■ Definition ■ Methods ■ Principle ■ Objectives ■ Centrifuge ■ Types and composition ■ Applications ■ Instruments ■ Super centrifuge

3. What is centrifugation? ☆ centrifugation is the process which involves the use of centrifugal force for the sedimentation of heterogeneous mixtures with centrifuge. ☆ centrifuge is used to separate particles (or) even macro-molecules such as: ● cells ● lipoprotein ● nucleic acids ● subcellular compounds ● proteins

4. METHODS OF CENTRIFUGATION.

5. DIFFERENTIAL CENTRIFUGATION Differential centrifugation (also known as Differential velocity centrifugation ) is a Very common procedure in biochemistry and cell biology which is used to separate organelles and other sub-cellular particles based on their sedimentation rate. Example: ◇ nanoparticles ◇ colloidal particles ◇ viruses

6. DENSITY GRADIENT CENTRIFUGATION A procedure for separating particles (such as viruses or ribosomes or molecules such as DNA) in which the sample is placed on a preformed gradient as sucrose or caesium chloride. Upon centrifugation either by rate zonal or equilibrium procedures,the macromolecules are “banded” in the gradient and can be collected as a pure fraction. Density gradient centrifugation is of two types: 1.rate zonal 2.isopycnic

7. RATE ZONAL CENTIFUGATION * The sample is applied in a thin zone at the top of the centrifuge tube on a density gradient. * Under centrifugal force ,the particles will begin sedimenting through the gradient in separate zones according to their size,shape,density or the sedimentation co-efficient. * The run must be terminated before any of the separated particles reach the bottom of the tube. Note: This method is useful fir separating particles which differ in size but not in density.Extremely useful for the separation of proteins possessing nearly identical densities but differing only slightly in their molecular weights.

8. ISOPYCNIC CENTRIFUGATION In isopycnic separation , also called buoyant or equilibrium separation, particles are separated solely on the basis of their density . Particle size only affects the rate at which particles move until their density is the same as the surrounding gradient medium.

9. PRINCIPLE OF CENTRIFUGATION The centrifugation involved in the principle of sedimentation.The particle having more than 5 micrometer are separated by sediminate under gravity process,the particle having size 5 micrometer or less ,do not sediminate under the rarity to centrifugal force is used to separate them. F=mv²/r _ _ _ _ _ (1) Let us consider a body of mass(m) rotating in a circular path with radius(r) at a velocity of (v). The force acting on the body is routine direction given by The gravitational force acting upon the same body is given as, G=mg _ _ _ _ _ (2) where, G= gravitational force g= acceleration due to gravity

10. The centrifugal effect (c) or relative centrifugal force is the ratio of the centrifugal force (F) and gravitational force (G) is expressed as C=F/G _ _ _ _ _ _ (3) C=v²/gr _ _ _ _ __(4) By substituting the value of v=2πrn in above equation, C=(2πrn)²/gr = 4π²rn²/g _ _ _ _ _(5) where, n= speed of rotating 2r=d(diameter of rotating) By putting the volume in above equation yield C=2π²dn²/d The value of gravitation is 9.807m/s² Centrifugal force = 2.013n²d RCF=11.18×R× [rpm/1000]²

11. OBJECTIVES OF CENTRIFUGATION ■ To separate the immiscible liquids. ■ To purify the component by removing impurities in the supernatant liquid. ■ To separate crystalline drugs from the mother liquor. ■ To test the emulsion and suspension for creaming and sedimentation at a accelerated speed.

12. CENTRIFUGE A centrifuge is a device that uses centrifugal force to separate various components of a liquid. This is achieved by spinning the fluid at high speed within a container,thereby separating fluids of different densities( e.g. cream from milk) or liquids from solids.

13. COMPOSITION ■ Centrifuge consists of four parts: * Drive and speed * Overspeed systetem * Temperature control * Vacuum system * Rotors Drive : water- cooled electric motor Speed control : 1. Selected hy rheostat 2. Monitored with a tachometer

14. Overspeed system : ■ Prevent operation of a rotor above its maximum rated speed ■ consists of : 1 . A ring of alternating reflecting and nonreflecting surfaces attached to the bottom of the rotor. 2. A small but intense point of source of light 3. A photo cell

15. Temperature control: ■ High speed centrifuge: ~~placing a thermocouple in the rotor chamber ~~monitoring only the rotor chamber temperature ■ Ultra centrifuge : ~~an infrared radiometric sensor placed beneath the rotor ~~continuously monitors the rotor temperature Vacuum system: ■ The speed of centrifuge < 15000 to 20000 rpm ■ The speed of centrifuge >4000rpm [phone]

16. TYPES OF CENTRIFUGE Ultra centrifuge High speed Centrifuge Desktop centrifuge

17. DESKTOP CENTRIFUGE ♤ Very simple and small ♤ Maximum speed of 3000rpm ♤ Do not have any temperature regulatory System. ♤ Used normally to collect rapidly sedimenring substances such as blood cells,yeast cells or bulky precipitates of chemical reactions.

18. HIGH SPEED CENTRIFUGES ■ Maximum speed of 25000rpm,providing 90000gm centrifugal forces. ■ Equipped with refrigeration to remove heat generated. ■ Temperature maintained at 0-4°c by means of the thermocouple. ■ Used to collect microorganism, cell debris, cells, large cellular organelles, precipitates of chemical (nuclei,mitochondria,lysosomes).

19. ULTRA CENTRIFUGE ■ Operate at speed of 75000 ram,providing the centrifugal force of 500,00gm. ■ Rotor chamber is sealed and evacuated by pump to attain Vacuum. ■ Refrigeration system(temp 0-4°c) ■ Rotor chamber is always enclosed in a heavy armor plate

20. APPLICATIONS. ♧ To separate two missile substances. ♧ Purification of mammalian cells. ♧ To analyze the hydronic properties of macromolecules. ♧ Fractions of subcellular organelles (including membranes / membrane fractions) . Fractionation of membrane vesicles.

21. INSTRUMENTS 1.Perforated Basket Centrifuge 2.Non perforated Basket Centrifuge 3.Semi continuous centrifuge 4.Super Centrifuge

22. SUPER CENTRIFUGE Principle: Super centrifuge is a type of sedimentation in which separation is achieved based on the density difference between two immiscible liquid. Construction: It contains small diameter cylindrical bowl . The Bowl is separatable, the bottom of bowl for the feed inlet using crushing system. At different height at the top of the bowl two liquid outlet are collected the separated liquid.

23. Working: The mortar rotation high frequency of about 2000rpm the liquid phase separated during centrifugation with the liquid phase separate during centrifugation with the heavy liquid and light liquid forming an inner layer. The liquid- interface (neutral zone) maintain the hydrophilic balance. The two layer of separating liquid to super centrifuge involve in the continue separation of liquid phase. APPLICATION: ☆ It is used for cleaning liquid , separation of clear liquid from suspension , emulsion. ☆ It is used for clarification of oil , fruit juice.

24. ☆Thankyou☆