CLINICAL CHEMISTRY

LECTURE
CLINICAL CHEMISTRY
CLINICAL CHEMISTRY
 Clinical chemistry (also known as clinical
biochemistry, chemical pathology, medical
biochemistry or pure blood chemistry) is the area of
pathology that is generally concerned with analysis of
bodily fluids.
 Late 19th century
ROLE OF MEDICAL TECHNOLOGIST
 Electrolytes
 Blood gases
 Enzymes assays
 Toxicology
 Lipid screening
 Therapeutic drug monitoring & Protein
 Quality control
 Hormone testing
VARIABLES AFFECTING CHEMISTRY ANALYSIS


UNITS OF MEASUREMENT

SI CONVERSIONS
To convert between SI units, move the decimal the difference
between the exponents represented by the prefix of the base
unit.
 If converting from smaller unit to larger unit, then
move decimal to the left the exponent difference.
 If converting from larger unit to smaller unit, then
move decimal to the right the exponent difference
Example 1: Convert 1.0 L to μL 1.0 L (1 × 10 0 ) μL
(micro = 10 –6 )
The difference between the exponents = 6.
The conversion is from a larger unit to a smaller unit,
so the decimal will move 6 places to the right.
1.0 L = 1,000,000 μL

REAGENTS

CHEMICALS

Analytic chemicals exist in varying grades of purity:

 Reagent grade or analytic reagent (AR);
- suitable for use in most analytic laboratory
procedures.

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 - The labels on reagents should clearly state the  instrument feed water,
actual impurities for each chemical lot or list the  water supplied by method manufacturer,
maximum allowable impurities.  autoclave and wash water,
- label should also include one of the following  commercially bottled purified water.
designations: AR or ACS or For laboratory use or
ACS Standard Grade Reference Materials. LAB WATER GRADES
 Type 3
 ultrapure, - lowest grade
- chromatography, - glassware rinsing
- immunoassays, - heating baths and filling autoclaves
- molecular diagnostics, - feed type 1 lab water systems
- standardization,  Type 2
- other techniques that require extremely pure - general lab applications (buffers)
chemicals. - feed to type 1 water systems
- free of contaminants - prep of reagents for chem analysis or synthesis
- have designations of HPLC (high-performance liquid  Type 1
chromatography) or chromatographic on their labels. - reagent grade
- ultrapure
 chemically pure (CP); -HPLC mobile phase preparation
- impurity limitations are not stated, and preparation of - blanks and sample dilution in GC, HPLC
these chemicals is not uniform - other advanced analytical techniques
- pure grade  HPLC
- not use for reagent preparation High-performance liquid chromatography
(HPLC) is a broad analytical chemistry
 United States Pharmacopeia (USP); National technique used to separate compounds in a
Formulary (NF); chemical mixture.
- used to manufacture drugs
- not harmful to humans SOLUTION PROPERTIES
- potable:dilute  Solute
- substance that is dissolved in a liquid
 Analyte
REFERENCE MATERIALS - biologic solute
 Primary Standard  Solvent
- highly purified chemicals that can be measured - liquid in which the solute is dissolved
directly to produce a substance of exact concentration Together, solute and solvent represent a Solution.
and purity Substances found in biologic fluids, including
- 100 +/- 0.02% purity tolerance serum, plasma, urine, and spinal fluid, are quantified
 Secondary standard
- lower purity
WATER SPECIFICATION CONCENTRATION
 Water – most common reagent
 Distilled water PERCENT SOLUTION
- been purified to remove almost all organic materials - expressed as the amount of solute per 100 total units of
- by distillation solution.
 Deionized water  weight per weight (w/w),
- Ions removed but may still contain organic material. - number of grams of solute per 100 g of solution.
- Purified from treated water using ion-exchange resins.  volume per volume (v/v),
- Deionized water is produced using either an anion- or a - used for liquid solutes and gives the milliliters of
cation-exchange resin, followed by replacement of the solute in 100 mL of solution.
removed ions with hydroxyl or hydrogen ions. - recommended that grams per deciliter (g/dL) be
 Reverse osmosis used instead of % v/v.
- uses pressure to force water through a semipermeable  weight per volume (w/v).
membrane, producing a filtered product. - most commonly used percent solution
- pre-treatment of water - grams of solute in 100 mL of solution.
- separation of impurities MOLARITY (M)
 Filtration - expressed as the number of moles per 1 L of solution.
- can remove particulate matter from municipal water supplies - One mole of a substance equals its gram molecular weight
before any additional treatments. (gmw)
 Ultrafiltration - customary units of molarity (M) are moles/liter.
- ultraviolet radiation, sterilization, ozone treatment - Molarity depends on volume, and any significant physical
-Microfiltration, Ultrafiltration, Nanolization, Reverse osmosis changes that influence volume, such as changes in
 Water Quality Testing: temperature and pressure, will also influence molarity
- Tests include resistance, pH, bacterial contamination, and MOLALITY (m)
chemical levels (e.g., chlorine, ammonia).
- represents the amount of solute per 1 kg of solvent.
- Pure water has high resistance but resistance alone doesn't
- moles per kilogram
indicate full purity.
 Reagent-grade water is classified into six categories
NORMALITY
- number of gram equivalent weights per 1 L of solution
based on its intended use, not the purification
 An equivalent weight
method:
is equal to the gmw of a substance divided by its
 clinical laboratory reagent water (CLRW),
valence.
 special reagent water (SRW),

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  The valence pH – 0-14 0-7 -acidic
is the number of units that can combine with or 7 – neutral
replace 1 mole of hydrogen ions for acids and 7- 14 – basic
hydroxyl ions for bases and the number of electrons
exchanged in oxidation–reduction reactions. HENDERSON-HASSELBACH EQUATION
- always equal to or greater than the molarity of the compound. which mathematically describes the dissociation characteristics
- Chemical titrations and chemical reagent classification of weak acids (pKa ) and bases (pKb ) and the effect on pH:

A solution is considered saturated when no more solvent can

be dissolved in the solution.
 Dilute solution
- as one in which there is relatively little solute or one
that has a lower solute concentration per volume of SIGNIFICANT FIGURES
solvent than the original, such as when making a are the minimum number of digits needed to express a
dilution. particular value in scientific notation without loss of accuracy.
 Concentrated
- has a large quantity of solute in solution. There are several rules in regard to identifying significant
 Saturated solution figures:
- A solution in which there is an excess of undissolved 1. All nonzero numbers are significant (1, 2, 3, 4, 5, 6, 7, 8, 9).
solute particles. (more concentration) 2. All zeros between nonzero numbers are significant.
 Supersaturated 3. All zeros to the right of the decimal are not significant when
- even greater concentration of undissolved solute followed by a nonzero number.
particles than a saturated solution of the same 4. All zeros to the left of the decimal are not significant.
substance. Examples:
COLLIGATIVE PROPERTIES 814.2 has four significant figures
 Vapor pressure 0.000641 has three significant figures,
-is the pressure exerted by the vapor when the liquid 10.00 has four significant figures.
solvent is in equilibrium with the vapor.
 Freezing point PERCENT SOLUTION
-is the temperature at which the first crystal (solid) of solvent Percent implies “parts per 100,” which is represented as
forms in equilibrium with the solution. percent (%) and is independent of the molecular weight of a
 Boiling point substance.
- is the temperature at which the vapor pressure of the solvent
reaches atmospheric pressure (usually 1 atmosphere).
 Osmotic pressure
- is the pressure that opposes osmosis when a solvent flows
through a semipermeable membrane to establish equilibrium
between compartments of differing concentration.

REDOX POTENTIAL
- oxidation–reduction potential, is a measure of the ability of a
solution to accept or donate electrons.
 lose electrons oxidized
 gain electrons reduced
 donate electrons are called reducing agents
 accept electrons are considered oxidizing agents.
oxidizing agents gain electrons and are reduced, while
reducing agents lose electrons and are oxidized.

CONDUCTIVITY
a measure of how well electricity passes through a solution.
 Resistivity
- reciprocal of conductivity, is a measure of a substance’s
resistance to the passage of electrical current.
- assessing the purity of water.

pH AND BUFFERS
 Buffers
are weak acids or bases and their related salts that minimize
changes in the hydrogen ion concentration.
 Hydrogen ion concentration
is often expressed as pH.
the term pH represents the negative or inverse log of the
hydrogen ion concentration.

Mathematically, pH is expressed as where [H+ ] equals the

concentration of hydrogen ions in moles per liter (M).

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  Bases: The valence is the number of hydroxide ions
(OH⁻) a base can donate. For example, sodium
hydroxide (NaOH) donates 1 OH⁻ ion, so its valence
is 1.
 Salts: For salts, the valence depends on the reaction
they undergo. For example, sodium chloride (NaCl)
has a valence of 1 because it forms a 1:1 ratio in
reactions.

MOLARITY
- expressed in units of moles per liter (mol/L) or sometimes
millimoles per milliliter (mmol/mL).
- 1 mol of a substance is equal to the gram molecular weight
(gmw) of that substance. DILUTIONS
represents the part(s) of concentrated material to the total final
volume of a solution.
ratio of concentrated or stock material to the total final volume
of a solution and consists of the volume or weight of the
concentrate plus the volume of the diluent, with the
concentration units remaining the same.
-Dilution factor
Ratio of consecrated or stock solution to the total solution
volume
A ratio of the concentrate to the lotal (final) volume.
A 1.4 dilution has a 1 volume of sample and 3 volumes of
diluent mixed together.
Any volume can be used to create this dilution, but It must be
the same unit of volume
Keep in mind the sample size when making your dilution
For example: a 2:3 dilution could contain
2 ml serum: 1 ml pure water
20 µl of serum 10 µl of pure water
0.2 mL of serum: 0.1 mL of pure water

NORMALITY

How do you find the valence electrons? For neutral atoms, the
number of valence electrons is equal to the atom's main group
number. The main group number for an element can be found
from its column on the periodic table. For example, carbon is in
group 4 and has 4 valence electrons. The number of valence
electrons for any atom is equal to the number of electrons in its
outermost shell. Determine the Equivalent Factor:
 Acids: The valence (or equivalent factor) is the
number of protons (H⁺ ions) an acid can donate. For
example, sulfuric acid (H₂SO₄) can donate 2 protons,
so its valence is 2.

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 of the scientific notation expression

pH AND POH
[H ^ n] = acid concentration
• [OH] = base concentration
• [H^ + ]*[OH^ - ]= 1 * 10 ^ - 14
• pH + pOH = 14

SERIAL DILUTIONS
Serial dilution involves making multiple, progressive dilutions to
reduce the concentration of a highly concentrated solution.

LOGARITHMS
inverse of exponential functions and can be related as such:
- Character
number to the left of the decimal point in the log and is derived
from the exponent
Mantissa
right of the decimal point and is derived from the number itself

x is negative exponent base 10 expressed without the minus

sign
N is the decimal portion

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