1. Chapter Clinical Laboratory Chemistry Copyright ©2011 by Pearson Education, Inc. All rights reserved. Clinical Laboratory Chemistry Sunheimer Graves Laboratory Basics 1

2. Copyright ©2011 by Pearson Education, Inc. All rights reserved. Clinical Laboratory Chemistry Sunheimer Graves Objectives Level I 1.Identify two methods used to produce clinical laboratory-grade water for use in the clinical laboratory. 2.List three items that should be monitored during the water-purification process. 3.Identify four types of glassware available for laboratory use.

3. Copyright ©2011 by Pearson Education, Inc. All rights reserved. Clinical Laboratory Chemistry Sunheimer Graves Objectives Level I 4.Identify four types of plastics used in laboratory plasticware. 5.Define the following terms: to contain (TC) and to deliver (TD) in reference to types of pipettes, molarity, molality, normality, thermocouple, percent solution, and hydrates.

4. Copyright ©2011 by Pearson Education, Inc. All rights reserved. Clinical Laboratory Chemistry Sunheimer Graves Objectives Level I 6.Cite three types of balances used to weigh substances in the laboratory. 7.Complete the mathematical calculations presented in this chapter correctly. 8.Convert results from one unit format to another. 9.Calculate the volumes required to prepare1:2, 1:5 and a 1:10 dilution.

5. Copyright ©2011 by Pearson Education, Inc. All rights reserved. Clinical Laboratory Chemistry Sunheimer Graves Objectives Level II 1.Explain the difference between air- displacement and positive-displacement micropipettes. 2.Distinguish swinging-bucket, fixed-angle- head, and ultra centrifuges from one another. 3.Explain the usefulness of liquid-in-glass, total-immersion, and partial-immersion thermometers.

6. Copyright ©2011 by Pearson Education, Inc. All rights reserved. Clinical Laboratory Chemistry Sunheimer Graves Objectives Level II 4.Identify three alternative thermometers that do not contain mercury. 5.Distinguish density, specific gravity, and assay by weight from one another. 6.Identify a source of calibration material for balances, thermometers, and pipettes.

7. Copyright ©2011 by Pearson Education, Inc. All rights reserved. Clinical Laboratory Chemistry Sunheimer Graves Water Purification Reverse Osmosis –Process by which H2O is forced through a semipermeable membrane that acts as a molecular filter Distillation –Process by which a liquid is vaporized and condensed to purify or concentrate a sustance or separate volatile substances from less-volatile substances Resistivity –Is the electrical resistance in ohms measured between opposite faces or a 1.00-cm of an aqueous solution at a specified temperature. –Used to assess the ionic content of purified water –↑ ion concentration, ↓ resistivity

8. Copyright ©2011 by Pearson Education, Inc. All rights reserved. Clinical Laboratory Chemistry Sunheimer Graves Laboratory Chemicals Chemical grades –ACS Reagent Analytical Pharmaceutical grades –The United States Pharmacopoeia (USP) –The National Formulary and The Food Chemical Index

9. Copyright ©2011 by Pearson Education, Inc. All rights reserved. Clinical Laboratory Chemistry Sunheimer Graves Laboratory Chemicals IUPAC –Grades A – E National Institute of Standard and Testing (NIST)

10. Copyright ©2011 by Pearson Education, Inc. All rights reserved. Clinical Laboratory Chemistry Sunheimer Graves Laboratory Glassware and Plasticware Glassware –Borosilicate (High degree of thermal resistance, low alkali content, free of heavy metals) –Low actinic (High thermal resistance, amber or red color protect light sensitive materials) Plasticware –Polypropylene (plastic pipette tips) –Polystyrene (capped graduated tubes, test tubes) –Polycarbonate (tubes for centrifuge, graduated cylinders, flasks; -100 to +160C) Teflon (stirring bars, tubing, cryogentics vials, bottle cap liners)

11. Copyright ©2011 by Pearson Education, Inc. All rights reserved. Clinical Laboratory Chemistry Sunheimer Graves Figure 1-1 Types of glass containers. A. Transparent glass; B. low actinic glass.

12. Copyright ©2011 by Pearson Education, Inc. All rights reserved. Clinical Laboratory Chemistry Sunheimer Graves Volumetric Laboratoryware Pipettes –TD (to deliver) –TC (to contain) –Micropipettes (air-displacement and positive-displacement) –Pipette calibration (accuracy&precision; weighting of water) Volumetric flasks –Calibration of volumetric flasks

13. Copyright ©2011 by Pearson Education, Inc. All rights reserved. Clinical Laboratory Chemistry Sunheimer Graves Figure 1-2 Several examples of glass pipettes. A. 0.2 mL TC; B. 1.0 mL TD serologic (blowout); C. 2.0 mL TD volumetric; and D. 10.0 mL TD Mohr. Note the two frosted- or etched-glass rings on pipette B.

14. Copyright ©2011 by Pearson Education, Inc. All rights reserved. Clinical Laboratory Chemistry Sunheimer Graves Figure 1-3 Several examples of micropipettes used in clinical laboratories.

15. Copyright ©2011 by Pearson Education, Inc. All rights reserved. Clinical Laboratory Chemistry Sunheimer Graves

16. Copyright ©2011 by Pearson Education, Inc. All rights reserved. Clinical Laboratory Chemistry Sunheimer Graves Balances Types –Unequal-Arm Substitution Balances –Magnetic Force Restoration Balance –Top-Loading Balances –Electronic Balances Calibration

17. Copyright ©2011 by Pearson Education, Inc. All rights reserved. Clinical Laboratory Chemistry Sunheimer Graves Figure 1-4 ASTM standard weight set used to calibrate laboratory balances.

18. Copyright ©2011 by Pearson Education, Inc. All rights reserved. Clinical Laboratory Chemistry Sunheimer Graves Centrifuges Relative centrifugal force and revolutions per minute Types –Swinging bucket centrifuge –Fixed angle rotors –An ultracentrifuge –Refrigerated centrifuge Maintenance and calibration

19. Copyright ©2011 by Pearson Education, Inc. All rights reserved. Clinical Laboratory Chemistry Sunheimer Graves Figure 1-5 Schematic showing the three major components of a typical laboratory centrifuge.

20. Copyright ©2011 by Pearson Education, Inc. All rights reserved. Clinical Laboratory Chemistry Sunheimer Graves Figure 1-6 Examples of two types of laboratory centrifuges. A. Swing-out rotor with buckets.

21. Copyright ©2011 by Pearson Education, Inc. All rights reserved. Clinical Laboratory Chemistry Sunheimer Graves Figure 1-6 (continued) Examples of two types of laboratory centrifuges. B. fixed-angle rotor.

22. Copyright ©2011 by Pearson Education, Inc. All rights reserved. Clinical Laboratory Chemistry Sunheimer Graves Water Baths Types –Circulating –Non circulating Maintenance and quality control

23. Copyright ©2011 by Pearson Education, Inc. All rights reserved. Clinical Laboratory Chemistry Sunheimer Graves Mixing Types –Single-tube mixers –Multiple-tube mixers

24. Copyright ©2011 by Pearson Education, Inc. All rights reserved. Clinical Laboratory Chemistry Sunheimer Graves Figure 1-7 Two types of tube mixers. A. vortex mixing.

25. Copyright ©2011 by Pearson Education, Inc. All rights reserved. Clinical Laboratory Chemistry Sunheimer Graves Figure 1-7 (continued) Two types of tube mixers. B. rocking motion of single and multiple tubes.

26. Copyright ©2011 by Pearson Education, Inc. All rights reserved. Clinical Laboratory Chemistry Sunheimer Graves Thermometry Types –Mercury free thermometers Organic red-spirit and pressurized with nitrogen gas Blue biodegradable liquid (isoamyl benzoate and dye) Red liquid thermometer filled with kerosene Bimetal digital thermometers Digital thermometers with stainless steel stems –Partial immersion thermometers –Full immersion thermometers

27. Copyright ©2011 by Pearson Education, Inc. All rights reserved. Clinical Laboratory Chemistry Sunheimer Graves Thermometry Thermistor Thermocouple Thermometer calibration

28. Copyright ©2011 by Pearson Education, Inc. All rights reserved. Clinical Laboratory Chemistry Sunheimer Graves Figure 1-8 Four examples of commonly used temperature monitoring devices. A. Mercury-filled partial immersion thermometer; B. red-spirit–filled partial-immersion thermometer; C. red-spirit–filled, total-immersion refrigerator thermometer.

29. Copyright ©2011 by Pearson Education, Inc. All rights reserved. Clinical Laboratory Chemistry Sunheimer Graves Figure 1-8 (continued) Four examples of commonly used temperature monitoring devices. D. digital type with thermocouple.

30. Copyright ©2011 by Pearson Education, Inc. All rights reserved. Clinical Laboratory Chemistry Sunheimer Graves Laboratory Mathematics Systems of measurement –Metric Length= meter Mass = gram Volume =liter Le Système International d’ Unitès

31. Copyright ©2011 by Pearson Education, Inc. All rights reserved. Clinical Laboratory Chemistry Sunheimer Graves Laboratory Mathematics Temperature –Celsius –Fahreheit –Kelvin Pressure –PO 2 and PCO 2 –Torr; Pascal

32. Copyright ©2011 by Pearson Education, Inc. All rights reserved. Clinical Laboratory Chemistry Sunheimer Graves Metric Prefixes PrefixAbbreviationFactor Femtof10 -15 Picop10 -12 Nanon10 -9 Microµ10 -6 Centic10 -2 Decid10 -1 Decada10 1 Hectoh10 2 Kilok10 3 MegaM10 6 GigaG10 9

33. Copyright ©2011 by Pearson Education, Inc. All rights reserved. Clinical Laboratory Chemistry Sunheimer Graves The Base Units of the Le Système International d’ Unitès QuantityNameSymbol LengthMeterM MassKilogramKg TimeSeconds Electric currentAmpere A Thermodynamic temperature KelvinK Amount of substanceMoleMol Luminous intensitycandelaCd

34. Copyright ©2011 by Pearson Education, Inc. All rights reserved. Clinical Laboratory Chemistry Sunheimer Graves Solution Concentrations Dilutions Density; Specific Gravity; assay by weight Atomic Mass Unit; Moles Molarity Normality Percent solutions Hydrates Enzymes

35. Copyright ©2011 by Pearson Education, Inc. All rights reserved. Clinical Laboratory Chemistry Sunheimer Graves Acids and Bases pH Buffer solutions