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TAGUM DOCTORS COLLEGE, INC.
Mahogany St., Rabe Subd., Visayan Village, Tagum City
BACHELOR’S IN MEDICAL LABORATORY SCIENCE
PRELIM
HANDOUTS
SUBJECT: CLINICAL CHEMISTRY LABORATORY
TOPIC: BASIC PRINCIPLES AND PRACTICE OF CLINICAL CHEMISTRY
NAME:
PLASTICWARES
POLYSTYRENE • Rigid clear, should not be autoclaved
• Will crack and splinter
• Used for test tubes, graduated tubes
POLYETHYLENE • Relatively inert chemically
• Resistant to most acids (except concentrated H2SO4), alkali and salts
• Used for test tubes, bottles, disposable transfer pipets, test tube racks
• Should not be autoclaved
POLYPROPYLENE • Relatively inert chemically
• Resistant to most acids, alkali and salts
• Can be autoclaved
• Used for pipette tips and test tubes
TEFLON • Almost totally chemically inert and resistant to a wide range of temperature
POLYCARBONATE • Very clear and ideal for graduated cylinder
POLYVINYLCHLORIDE • Soft and flexible materials used frequently to construct tubing
POLYOLEFIN • Noted for their strength and resistance to elevated temperature
• Some determinations requires plastic rather than glass because it can absorb metal ions
GLASSWARES
HIGH THERMAL • High resistance to thermal shock and chemical shock
• Heavy wall to minimize breakage
BOROSILICATE • Used for most beakers, flasks and pipets
(PYREX/KIMAX) • Minimal contamination of liquids by elements in glass
• Can be heated and autoclaved
HIGH SILICA • Acid and alkali resistant, heat and electrical tolerance
• Excellent optical pipettes
(VYCOR) • Used for high precision analytic work and optical reflectors and mirrors
LOW ACTINIC • Red or amber in color to prevent exposure of photosensitive light
FLINT GLASS • Soda-lime glass containing oxides of sodium, silicon and calcium
• Least expensive but poor resistance to high temperature and sudden changes of temperature
• Only fair resistance to chemicals
• Can release alkali and affect some determinations
• Used for some disposable glassware
ALUMINUM • Six (6) times stronger than borosilicate
SILICATE (COREX) • Better able to resists clouding due to alkali and scratching
HIGH THERMAL • Resist high temperature only
SOFT GLASS • Used for highly alkaline solutions, alkali resistant
BORON FREE • Poor heat resistance
LABORATORY VESSEL
VOLUMETRIC FLASK • Has round flat lower portion and a long thin neck with the calibration line etched into the neck
ERLENMEYER FLASK • Has a wide bottom that gradually evolves into a smaller, short neck
GRIFFIN BEAKER • Has A flat bottom, straight sides and an opening as wide as the flat base with a small spout in the lip
GRADUATED CYLINDER • Long cylindrical usually held upright by an octagonal or circular base
By: CLA, RMT
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● PIPETS
1. Utensils made of glass or plastic that are used to transfer liquids: may be reusable or disposable
2. clean by soaking to a soapy water with pipet tips up
3. used for volume of 20mL or less
4. with different tolerances depending on the volume
● CLASSIFICATION AND TYPES
1. DESIGN
2. DRAINAGE CHARACTERISTICS
3. TYPES (MEASURING/GRADUATED AND TRANSFER)
Tip should be immersed in the liquid to be transferred to a level that will allow it to remain in the solution after the volume of
DESIGN: liquid has entered the pipet.
TO CONTAIN • holds a particular volume but dies not dispense that exact volume
• delivers the amount by washing it and emptying
TO DELIVER • will dispense the volume indicated: transfer exact amount
• do not blow the last drop
• rate of flow is gravity
DRAINAGE CHARACTERISTICS
BLOW-OUT • ETCHED RING!!!!! BY THE MOUTH
SELF-DRAINING • NO ETCHED RING!!!!! DOOOOONT TOUCH TIP TO LIQUID
MEASURING OR GRADUATED TRANSFER
Capable of dispensing several different volumes Designed to dispense one volume without further subdivisions
Serologic (graduated up to the tip) Pasteur pipet
Mohr (has dead space) Volumetric pipet
Ball, Kolmer or Khan Ostwald Folin
Micropipet Micropipet/Automatic Pipet
By: CLA, RMT
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MICROPIPETTES ( mao na ni ginagamit nowadays sa lab class dli natung mga mhor, volumetric etc…)
H HOW TO USE MICROPIPETTES??
• Mechanical suction > wipe outside with gauze > adjust meniscus read at bottom
MENISCUS: The downward curve at the surface of a liquid.
PIPET CALIBRATION
GRAVIMETRIC METHOD - Done by delivering and weighing a solution of known specific gravity
such as water
- Most desirable method for pipet calibration
- Using analytical balance
PHOTOMETRIC METHOD - For automatic pipetting devices
- Absorbance of colored solution K dichromate or Paranitrophenol
- calibrate auto pipette
▪ BASIC SEPARATION TECHNIQUES
CENTRIFUGE – process whereby centrifugal force is used to separate solid matter from a liquid suspension
o Supernatant – liquid
o Sediment – solid
o Parts: HEAD/ROTOR, CARRIERS, SHIELDS
o Speed: Tachometer/ Strobe light
o Rule of thumb: Placing the tube by exact positioning or by the exact opposite
o Purpose of the hole in centrifuge machine: for speed verification
o RPM – revolution for minute
o RCF – relative centrifugal force
o s(Svedberg) – unit used for ultracentrifuge (lipoprotein analysis)
FILTRATION
o Used to separate serum from a blood clot used in place of centrifugation
o Filter materials is made of paper cellulose and its derivatives, polyester fibers, glass and a variety pf resin column
material
o Filter papers differs in terms of pore size and should be collected according to separation needs
o Filter paper should not be used when using strong acids or bases
o FILTRATE: liquid that passes through the filter paper
DIALYSIS – method for separating macromolecules from a solvent or smaller substance
By: CLA, RMT
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TYPES OF CENTRIFUGE
REFRIGERATED CENTRIFUGE 1. For thermally labile analytes
ULTRACENTRIFUGE 2. Separate layers of specific gravities
3. -15 to 20 degree Celsius
4. Rapid high speed
5. 100,000 RPM (actual speed)
6. Confirmatory for lipoproteins
FIXED ANGLE/ ANGLE HEAD 1. Capable of higher speed with much less heat build up
2. Decrease air friction/resistant
3. 25 – 40-degree angle
4. Max: 7,000 RPM only
5. 9,000 RCF (force acting on the sample)
SWINGING BUCKET 1. Spinning horizontal, vertical when at rest
2. For slow flat tightly packed sediments
3. Increase air friction/resistant
4. 1,650 RCF (force acting on the sample)
5. 3,000 RPM (actual speed)
CYTOCENTRIFUGE 1. Uses high torque
2. Low inertia motor to separate monolayers of cells for morphologic studies (body fluids analysis)
3. 200 – 2000 RPM (actual speed)
CLEANING OF GLASSWARES AND PLASTICWARES
ROUTINE WASHING may be done by: a. Soaking in dilute bleach followed by drying in an oven
b. Soaking in 20% nitric acid solution for 12-24hrs
c. Soaking in ACID DICHROMATE SOLUTION
FOR BLOOD CLOTS Soak in 10% NaOH
FOR NEW PIPETS Soak in 5% HCl or 5% HNO3
FOR METAL ION DETERMINATION Soak in 20% HNO3
FOR GREASE Soak in any organic solvent or 50% KOH or contrad 70
FOR PERMANGANATE STAINS Soak in 50% HCl or a mixture of 1% FeSO4 in 25% H2SO4
FOR BACTERIOLOGIC GLASSWARES Soak in 2% to 4% cresol solution followed by autoclaving and thorough washing
FOR IRON DETERMINATION Soak in HCl solution or HNO3 solution followed by thorough washing
FIXED ANGLE CENTRIFUGE SWINGING BUCKET CYTOCENTRIFUGE
ULTRACENTRIFUGE
By: CLA, RMT
