CLINICAL CHEMISTRY
 assays or tests are done to determine
increased or decreased levels of chemicals in
the body (by comparing to normal values)
Biochemical in the body need Homeostasis
(balance) = Normal Values
 Biochemicals – chemicals that make up living
things; that are found in life
1. Carbohydrates
2. Proteins
3. Lipids
4. Nitric Acid
 chemical levels in our body have been
determined and these levels are called Normal
Values (NV)
a. Hypo – if the chemical analyte determined
in the test goes below the normal values
b. Hyper – if the chemical analyte determined
in the test goes beyond the normal values
 these results provide vital evidences of what
is causing the disease of a person
Examples:
 Glucose level (normal value = 5.6 – 6.4 mmol/L)
 Hyperglycemia – higher than the NV
 Hypoglycemia – lower than the NV
 Sodium level
 Hypernatremia
 Hyponatremia
 Potassium level
 Hyperkalemia
 Hypokalemia
To maintain quality results in Clinical Chemistry
procedures, tests are run simultaneously with
standards and controls.
a. Standards
- basis for the concentration of the test
- used as a constant variable to which the
tested analyte is computed and controls are
used to determine accuracy of the test
b. Controls
- Normal and Abnormal Controls to determine
the acceptability of the test
These provide a guide to determine whether the
test performed is reliable or not.
METHODS USED IN CLINICAL CHEMISTRY
ANALYSIS
1. Spectrophotometry
 spectro = visible/seperated light; photo =
light; metry = measurement
 make use of prisms or gratings to isolate a
narrow range of visible (400 – 700 nm) and
invisible light
 prism – separate different components
of white light
 each light has its own wavelength
 wavelengths – measured in
nanometers (nm)
Spectrophotometer
 equipment used in measuring concentration
of analyte
 uses the Beer-Lambert Law; states that the
loss of light intensity when it propagates in a
medium is directly proportional to intensity
and path length
 it means that the higher the
concentration of an analyte in a tube,
the higher its absorbance and the
lower is its transmittance
Cuvette
 Tungsten light – common source of light
 Incident light – light that strikes to the test tube
 Absorbed light – light that is blocked by
particles
 Transmitted light – light that passed through
the test tube
 this transmitted light is now detected by
photodetector or detector which convert
the intensity of light into concentration
 measures absorbance not transmittance
(T = 2-logA)
Performing Test with Quality
 minimum of five test tube
 test tube 1 = blank
 to zero the machine
 test tube 2 = normal control
 test tube 3 = abnormal control
 tube 2 & 3 to check the acceptability of
the test
 used predetermine concentrations
 test tube 4 = standard
 basis for the measurement of
concentration of the test
 standard serve as a divisor
 concentration of standard serve as a
multiplier
 test tube 5
 serum/plasma
2. Nephelometry
 opposite of turbidimetry, both use lens but
measures scattered light not the transmitted
light
 detection of light energy scattered or
reflected toward a detector that is not in the
direct path of the transmitted light
3. Turbidimetry
  similar to spectrophotometry, both
measures transmitted light but use lens
instead of prisms
 process of measuring the loss of intensity of
transmitted light due to the scattering effect
of particles suspended in it
 measures the amount of light that is pass
through (transmitted light), but it will
compute for the turbidity based on the
amount of light that is blocked
 Light is passed through a filter creating a
light of known wavelength which is then
passed through a cuvette containing a
solution

4. Atomic Absorption Spectrophotometry

 atomic absorption methods measure the
amount of energy absorbed by the sample
 can measure even smallest quantity/
concentration of analyte
 very sensitive method of elemental analysis
particularly trace metals, allowing the
determination of metals in a variety of
samples at the picogram level
 Hollow Cathode Lamp – instrument uses a
light source
 emits specific wavelengths of light that
are ideally only absorbable by the
analyte

5. Flame Emission Spectrophotometry

 light source is flame; used in electrolytes
and substances that emit color when burn
 based on the characteristic emission of light
by atoms of many metallic elements when
given sufficient energy, such as that
supplied by a hot flame
 burn the solution which gives different
colors
 Lithium = red flame
 Sodium = yellow flame
 Potassium = violet flame
 Magnesium = blue flame

6. Electrophoresis
 phoresis = movement
 general term that describes the migration
and separation of charged particles (ions)
under the influence of an electric field
 movement of substances depending on the
charges of particles called ions
 cations (positive charge) move towards the
cathode (negative electrode) while anions
(negative charge) move towards the anode
(positive electrode)
 the movement of substances is dependent
on the charges of analyte