CLINICAL CHEMISTRY: BASIC PRINCIPLES & PRACTICE - Calibrated for volume it delivers (more accurate)

2. Drainage
Clinical Laboratory Supplies  Self-draining
1. Thermometer (units, types) - Contents drain via gravity
 Thermistor – electronic  Blow-out
 Liquid-in-glass – alcohol with dye - Makes use of an aspirator bulb to blow out
2. Glassware contents
 Should be Class A - Has mark near mouth piece (top) of an etched ring
 Borosilicate / double ring / frosted band
- Brands: Pyrex, Kimax Types of Pipettes
- Incorporated with Boron Oxide 1. Measuring / Graduated
- Free from magnesia-lime-zinc group of elements  Dispenses several different volumes
(contaminants)  With many calibration / graduation lines
- High thermal resistance glassware  510 C  Measures several fractional volumes as long as
- Temperature transitioning should be slow calibrations suffice
- Not chemically strong (can’t handle very acidic /  Mohr (with residual liquid)
alkaline solutions) - TD, self-draining, calibrated between two marks
 Aluminosilicate  Serologic (full pipette)
- Acid and alkaline resistant - TD, blow-out, calibrated down to the tip (end)
- High thermal resistance 2. Transfer
- Chemically strengthened  Measure only one volume
- Brands: Corex (TR: 672 C), Vycor (TR: 900-1200 C)  Has a bulb on the stem
 High Silica  Volumetric
- Same thermal resistance as Vycor - Bulb in the middle, TD, self-draining, for aqueous
- Optically pure (clear glass) solutions
 Low Actinic  Ostwald Folin
- Glass w/ color (red or amber) - Bulb on lower part of stem, TC/TD, blow-out, for
- For vitamins, metabolites, bilirubin, and other viscous solutions
photosensitive substances 3. Pasteur
 Flint  Droppers
- Soda-lime glass 4. Automatic
- Thinnest material and least expensive  Air Displacement
- For making disposable glassware (droppers) - Principle of operation: air displacement
3. Plasticware - Air is aspirated and pushed out
 Flexibility, resistance to corrosion & breakage - Has disposable delivery tip per solution to avoid
 Can’t withstand high temperature contamination
 Polysterene, polyethylene, polypropylene, tygon,  Positive Displacement
Teflon, polycarbonate, PVC - Very much like needles
 Teflon - Found on machines
- Polycarbon - Machine washes itself every time tests are run,
- TR: 255 C thus washes the pipettes as well
- Used for extremes of temperature  Dispenser or Diluter / Dispenser
- Can be frozen to temperatures as low as -270C - Has a container for dilution + capillary pipette
 Cleaning 5. Micropipette
- General use: soapy water, acid dichromate  Small capacity
mixture  Transfer of 1ml or less
- Blood clots: 10% NaOH  1 ml = 1000 µm
- New pipettes: 5% HCL; 5%HNO3  Ex: RBC and WBC pipettes (graduated)
- Metal ion determinations: 20% HNO3; 1:2 HNO3;
1:2 HCl Reagents
- Grease: Organic solvent; KOH & Ethanol solution; 1. Chemicals
Contrad 70  Analytical procedures are prone to contaminations
- Bacteriologic: 2-4% ethanol  Grades: Analytical Reagent Grade, Ultrapure, Chemically
- Permanganate stains: 50% HCl; 1% Ferrous Pure, United States Pharmacopoeia, National Formulary,
Sulfate in 25% H2SO4 technical / commercial
4. Laboratory Vessels  Analytical Reagent Grade
 Flasks, beakers, graduated cylinders - Certified by the American Chemical Society
5. Pipettes (checks purity of chemicals)
 Transfer of liquids; glass or plastic; TC or TD; may or may  Ultrapure
not be blown out; may or may not be rinsed - Higher purity than ARG
6. Burettes - For special procedures: chromatographic grades
7. Balances  Chemically Pure
8. Desiccators / Desiccants - Not appropriate for analytical use
- Not certified (self-certification)
Classifications of Pipettes - Purity is checked by boiling / melting point analysis
1. Calibration / design - Variable checking of purity
 TC (to contain) or rinse-out  United States Pharmacopoeia and National Formulary
- Calibrated for the volume it contains - Safety for human consumption (manufacturing of
- Residual liquid changes final volumes when drugs)
transferred
 Technical / Commercial
- Rinsing technique: to increase accuracy
- Least purity
 TD (to deliver) - Not used for analytical procedures
 - Available commercially
2. Reference Materials Basic Separation Techniques
 Aka “standards” (have known conc.) 1. Extraction
 All lab tests are based on standards (calibrations)  Done in research labs (not clinical)
 Primary Standards  Uses immiscible solvents to extract
- Highest purity level 2. Lyophilization
- ACS certified (almost 100% purity)  Freeze-drying
- Expensive  Exposes solution/substance to sub-zero temp. to dehydrate
 Secondary Standards and conserve
- Not as expensive  Becomes powder in form which is reconstructed with a
- Values are based on primary standards solvent
 Standard Reference Materials / Certified Reference  Ex: powdered milk
Materials 3. Dialysis
- Routinely used in the lab  Uses dialyzing membrane to separate small particles from
- NIST (National Institute of Standards and bigger ones in a solution
Technology) certified 4. Filtration
5. Centrifugation
Water: most commonly used reagent in the laboratory (reagent grade water)  Used in the clinical chemistry lab
 For obtaining serum and plasma
Types of Water (based on purity level)  Uses gravitational force to separate blood components based
1. Type 1 Water on density
- Most pure  RCF = relative centrifugal force: 1.118 x 10-5 x r x rpm2
- Minimal contaminants  Tachometer: inducts speed and force of centrifuge
- For preparing standards  Rotor: spinning part with buckets / carriers
- Cannot be stored because of contamination  Carriers: where test tubes are placed
possibilities
 Standard: 10 minutes; 850-1000 x g
2. Type 2 Water
- Good for most lab procedures
Types of Centrifuge
- Storage: short duration only
1. Refrigerated or Unrefrigerated
3. Type 3 Water
2. Floor type: big
- Used for qualitative procedures
3. Bench type: small
- For glassware washing
4. SeroFuge: calibrated to use for serum
5. Ultra-Centrifuge: based on speed (faster)
Type 1 Type 2 Type 3 6. Swinging Bucket / Horizontal Head
Max. Colony Count <10 1000 NS (not - Movable, horizontally inclined, uniform sediment
-bacterial contam. specified) production
-colony forming units per mL 7. Fixed / Angle Head or Fixed Head
Silicate 0.05 0.1 1.0 - Angled carriers at 15-40°, non-movable, subjected
-glass particles (contamination) to more air friction, slanted
-mg/L
Resistivity 10 1.0 0.1
-inverse of conductivity to
electricity
-the purer the water, the less
conductive
-megOhms/cm
Ph NS NS 5-8
(neutral) (neutral)

Methods of Water Purification

1. Distillation
- Oldest method
- Vaporization & condensation
- Removal of organic contaminants
2. Deionization
- Removal of ion impurities
- Principle involved: ion exchange via resins
- Anion Exchange Resins: remove anionic
impurities in exchange for OH ions
- Cation Exchange Resins: remove cationic
impurities in exchange for H ions
- Mixed Bed Resins: both anionic and cationic
exchange
3. Reverse Osmosis
- Water forced to pass through semi-permeable
membrane
4. Filtration
- Submicron filtration
- Filter pore size: <0.22 µm
5. UV Light and Ozone Treatment
- Removal of microorganisms
6. Sterilization: autoclave