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CHEMISTRY
Julius P. Mario, RMT, MS Chem.
2.
3.
Establishing a prognosis
4.
5.
Clinical Chemistry
The systematic study of
biochemical processes associated
with health & disease &
the measurement of constituents in
body fluids or tissues to
facilitate diagnosis of disease.
Instrumentation
CLINICAL CHEMISTRY
Pharmacology
Immunology
Endocrinology
Toxicology
Analytical Chemistry
Point-of-care Testing
(POCT)
Also known as near-patient testing, alternate-site
testing or patient-focused testing
Specimens or samples
are analyzed
while
substances in them are
measured or quantitated.
2.
glucose
electrolytes
TP
TAG, cholesterol
albumin
hormones
individual proteins
vitamins
creatinine
uric acid
bilirubin
3.
4.
antibodies
therapeutic drugs
substances of abuse
poisons
Quick Quiz
Laboratory Tests
Chemical tests
Direct or Indirect
Colorimetric
Redox
Condensation
Degradative tests (Enzymatic)
One-step or Multi-step
NADH Consumption
Measurement of Activity (not
concentration)
Multi-method
Titrimetric
Electrochemical
Quick Quiz
LAB REQUESTS
Glasswares
Plasticwares
Highly shock proof;
shatterproof
Breakable
Reusable
Disposable
1.
Corex
Vycor
reagent bottles
and
disposable laboratory glasswares
Quick Quiz
PLASTICWARES
1.
Polyolefins (polyethylenes,
polypropylenes)
Unique group of resins with relatively inert
chemical properties
Polypropylene is sterilizable
however it absorbs pigment and tends to
become discolored
PLASTICWARES
2. Polycarbonate resin
tubes and
graduated cylinders
3. Tygon
tubings
Flexible
Quick Quiz
PIPETS
Two General Types
1.
liquid
Consists of a cylindrical
narrower glass tubing
measurements of aliquots
of nonviscous samples, filtrates, controls,
and standard solutions
OSTWALD-FOLIN PIPET
Mohr pipet
- calibrated between two marks on
the stem
Serological pipet
- has graduation marks down on
the tip and blown out to deliver the entire
volume of the pipet if etched ring or double
rings are evident near the mouth of the pipet
Serological pipet
MICROPIPETS
Sahli (TC)
Kirk (TC)
Lang-Levy (TD/ TC)
Overflow (TC)
Capillary (TC)
Sanz Pipet (TD=TC)
Unopette (TC)
Semiautomatic pipet Eppendorf (TD/TC)
Quick Quiz
CHEMICALS
spectrograde,
nanograde, and
STANDARDS
Quick Quiz
WATER
Reagent grade water (RGW) is water
Type I
Type II
Type III
<10
102 NA
NA NA
0.05
0.1
0.1 1.0
activated carbon
NA
NA
NA
NA
Quick Quiz
Chemical Hazards
Caustic can cause burns
Flammable also combustible, that is,
can
easily catch fire
Corrosive harmful to mucous
membranes,
skin, eyes or tissues
Carcinogen can produce or incite cancer
Mutagen cause changes in RNA or
DNA
Teratogen cause birth defects
Quick Quiz
CENTRIFUGE
The relative centrifugal force (RCF) is obtained by
using a nomogram or by using this formula:
strobe light.
pipetting is strictly
prohibited.
Quick Quiz
BIOLOGIC SAFETY
C.
D.
E.
F.
G.
routes of entry;
exposure limits;
carcinogenic potential;
emergency first-aid.
MSDSs contain information on the nature of the chemical,
precautions if spilled and disposal recommendations.
Special Considerations
Ox
Water Reactive
Oxidizing Agent
Radioactive
Poison
ELECTRICAL SAFETY
RADIATION SAFETY
Quick
Quiz
FIRE SAFETY
Fires have been divided into four classes on
the nature of the combustible material and
requirements for extinguishments.
Class A- ordinary combustible solid materials
such as paper,
wood,
plastic and rubber
Class B- flammable liquids/ gases and
combustible petroleum products
Class C- energized electrical equipment
Class D- combustible/reactive metals such as
Mg, Na and K
Class E nuclear reaction
Class K cooking media
Quick Quiz
burial
recycling
Biohazardous Wastes
Quick Quiz
UNITS OF MEASUREMENT
Basic Unit
Symbol
Length
meter m
Mass
kilogram
kg
Time
second s
Electric current
ampereA
Temperature
Kelvin K
Luminous intensity Candela
Amount of substance
mole
Catalytic amount
katal
cd
mol
Symbol
Conductance
Siemens
Resistance
Activity (radionuclide)
Becquerel
m3
Bq
Quick Quiz
EFFECT ON LABORATORY
RESULTS
Increased Decreased
Exercise
Immediate effects:
Alanine and lactate
FFA
Long-term effects
CK, LD, AST, platelets,
testosterone,
androstenedione and LH
Fasting Most analytes
(8-12 hrs for common
bilirubin, FFA, TAG
metabolites and
12-14 hrs for lipid profile)
profile)
Glucose
Increased
Decreased
After eating
K, TAG (chylomicrons),
turbidity of serum and plasma
Diet
High meat
NPNs (esp. urea, ammonia,
and urate except creatinine)
High ratio of unsaturated
to saturated fat
Purine-rich
Serum
cholesterol
Urates
Serotonin in blood
5HIAA in urine
Tobacco smoking
Acute effects
Chronic effects
Increased
Decreased
Catecholamines, cortisol
neutrophils, monocytes
and FFA
Eosinophil
Stress
Quick Quiz
BLOOD COLLECTION
Venipuncture (Phlebotomy)
movement techniques.
Sites of venipuncture includes:
Complications of venipuncture-classified as
immediate or delayed and local or systemic
Hematoma due to missed vein
Collapsed small veins due to excessive pull of
plunger
Syncope
Excessive bleeding
Thrombosis of vein
Infection is situ
Blood-borne infection like hepatitis B and
AIDS
Considerations
Quick
Quiz
Arterial puncture
Radial arteries
Brachial arteries
Femoral arteries
Considerations
consumption of oxygen.
Quick Quiz
Infants
Lateral plantar heel surface
Median plantar heel surface
Children
Plantar surface of the big toe
Plantar surface of the last digit of the 2 nd, 3rd or 4th fingers
may be used in older infants
Lateral side of the finger adjacent to the nail
Quick Quiz
VACUTAINER/EVACUATED TUBES
COLOR
ADDITIVE
Lavender
Ethylenediaminetetraacetic
chelates calcium
Acid (EDTA)
* Versene (disodium salt)
* Sequestrene (dipotassium salt)
Red
None
Red Gray or
Red Black
separator material
and serves as a
barrier between cells
and serum
Yellow
Citrate dextrose
Green
ACTION
preserves RBCs
inhibits thrombin
activation
USE
hematologic assays
lead assay, CEA
Determination and
cell counts
Most chemistry,
immunologic and
blood bank tests
Most chemistry tests
Blood culture
ammonia CO-Hb
and methemoglobin
COLOR ADDITIVE
Orange Thrombin
Blue
ACTION
USE
accelarated clot
Coagulation assays
binds calcium
Westergren ESR
citrate
Gray
NaF/K2C2O4
Inhibits glycolytic
enzyme enolase and
determination
act as anticoagulant
Iodoacetate inhibits glycolytic
3-phosphate
dehydrogenase
Glucose
Glucose
enzyme glyceraldehyde
determination
Anticoagulant Interference
Quick Quiz
Specimen Interference
collection
volume of specimen
Inappropriate
collection tube
Hemolysis
Improper
transportation
Interferences
QUALITY MANAGEMENT
Quality Assurance (QA)
of specimen
entering patient information
centrifuging
separating aliquoting specimen
delivery to proper laboratory location
involves
and precision
of control samples
instrument maintenance
statistical data analyses
proficiency testing survey
Quick
Quiz
Which
QC chart is a graphic representation
of the acceptable limits of variation in the
results of an analytical method?
a. Gaussian
b. Youden
c. Levey-Jennings d. Cusum
Errors
Analytical errors are usually systematic
errors or determinate errors that are
caused by some factors in the analytical
system such as erroneously calibrated
pipettor, deteriorating reagent and
fluctuating electrical current.
Personnel or operator errors are usually
called random errors or indeterminate
errors that usually affect several analyses.
Examples are mislabeling the specimen,
wrong number entry and instability of
needle due to electronic component of
instrument.
Shift
Dispersio
n
Trend
Quick Quiz
Westgard Multi-Rule
Technic
Rule
Meaning
1:2s = one control observation exceeds the control
limit set at
M2SD; warning rule
1:3s = reject when one observation exceeds M3SD;
suggests
random error
2:2s = reject when two consecutive observations
exceed the same
M+2SD or the same M-2SD;
suggests systematic
error
R:4s = reject when one control observation in the run
exceeds its
M+2SD and another exceeds M-2SD;
suggests
random errors
4:1s = reject when four consecutive control
observations exceed
the same M+1SD limit or
the same M-1SD limit; suggests
systematic error
10:Mean= reject when 10 consecutive control
observations fall on one side of the mean; suggests
systematic error
Quick
Quiz
What
does the preparation of a Levey-Jennings
Standard deviation =
(M x)2
n-1
Variance = (SD)2
Coefficient of variation =
100%
M
SD x
Quick Quiz
Quick Quiz
Quick Quiz
(standard)
Figures of Merit of
Analytical Methods
Major
1. Accuracy
2. Precision
3. Limit of detection
4. Applicable concentration
range or
linear range
5. Sensitivity
6. Selectivity
Minor
1. Speed
2. Ease and convenience
3. Skill required of the
operator
4. Cost of analysis and
availability of equipment
5. Per sample cost
End