CLINICAL CHEMISTRY LECTURE

PRELIMS | 2ND SEMESTER

2-E | PROF LACIA

○​ Calibrating medium:
INTRODUCTION mercury

★​ Area of chemistry that deals w/body DRAINAGE CHARACTERISTICS

★​ BLOWOUT
fluids like blood and serum
○​ w/etched rings
○​ Using an aspirator to blow
TYPES OF GLASSWARES out the remaining amount
★​ SELF DRAINING
○​ w/out etched ring
★​ BOROSILICATE GLASS ○​ Liquid is allowed to drain by
○​ Most common, made up of gravity
boron trioxide and silica
○​ Used for heating and TYPES
sterilization ★​ TRANSFER - transfer liquids
○​ Has high resistance to alkali ○​ Volumetric pipette: used for
○​ Brand (pyrex&kimax) non viscous fluid such as
water, a self draining pipette
★​ BORON-FREE GLASSWARE ○​ Ostwald folin: used for
○​ Has high resistance to heat viscous fluid such as blood
(blowout)
★​ COREX ○​ Pasteur pipette: disposable,
○​ Type of borosilicate glass for practice purposes
(aluminum silicate) ○​ Automatic
micro/macropipettes: most
★​ VYCOR commonly used in the
○​ Resistant to high laboratory
temperature
○​ Can be heated up to 900’c ★​ GRADUATED OR MEASURING -
exact amount
★​ FLINT GLASS ○​ Serological pipette:
○​ Temperature sensitive w/graduation to the tip
(marks)
PIPETTE ○​ Mohr pipette: opposite to
serological, w/out graduation
to the tip (self draining)
★​ Transfer liquid from one container to ○​ Bacteriologic pipette: used
other container in bacteriology, unique
○​ Ball, kolmer and khan: type
CALIBRATION MARKS/DESIGN of graduated and measuring
★​ TO DELIVER (TD) ○​ Micropipette: used when we
○​ Delivers the exact amount of transfer liquid w/less than
liquid into a container 1ml (<1ml)
○​ Calibrating medium:
distilled water
★​ TO CONTAIN (TC)
○​ Does not dispense the exact
amount, only holds the liquid

CLIN CHEM 1 GWYN 1

 CLINICAL CHEMISTRY LECTURE
PRELIMS | 2ND SEMESTER
2-E | PROF LACIA

○​ Should never be used in

REAGENTS AND SUPPLIES clinical laboratory setting

★​ ANALYTICAL REAGENT GRADE

REFERENCE MATERIALS
(AR)
○​ Important for quantitative
and qualitative analysis ★​ Used for calibration, quantification
○​ Used in most analytical and verification of analytical
laboratory procedure standards
○​ Meets the specifications
established by the american ★​ PRIMARY STANDARD
chemical society (ACS) ○​ Highly purified chemical that
○​ Use: for trace metal analysis can be measured directly to
and preparation of standards produce a substance of exact
known concentration and
★​ ULTRAPURE REAGENT purity
○​ Put through additional
purification steps ★​ SECONDARY STANDARD
○​ Use: chromatography, ○​ Lower purity
atomic absorption and
immunoassays
WATER SPECIFICATIONS
★​ CHEMICALLY PURE (CP) / PURE
GRADE
★​ Most frequently used reagent in the
○​ Preparation of these
laboratory
chemicals is not uniform
★​ Distillation process - involves
○​ Not recommended for
conversion of a liquid into a vapor,
research and analytical
subsequently condensed back to
chemistry, unless a reagent
liquid form
blank (used to measure
background interferences
★​ DISTILLED WATER
and contamination from the
○​ Solely purified by distillation
chemicals) is included
★​ DEIONIZED WATER
★​ UNITED STATES PHARMACOPOEIA
○​ Water purified by ion
(USP) and NATIONAL FORMULARY
exchange
(NF)
○​ Also called demineralization
○​ Approved for human
(removal of ions)
consumption
○​ May not be applicable for
★​ RO WATER
laboratory analysis
○​ Produced by reverse osmosis
○​ Use: drug manufacturing
which pumps water across a
semipermeable membrane ​
★​ TECHNICAL OR COMMERCIAL
GRADE
○​ Used primarily in
manufacturing

CLIN CHEM 1 GWYN 2

 CLINICAL CHEMISTRY LECTURE
PRELIMS | 2ND SEMESTER
2-E | PROF LACIA

THREE GRADES OF REAGENT WATER ★​ Desiccator and desiccants (used to

★​ TYPE 1 REAGENT WATER store hydroscopic substances)
○​ Used for test methods ★​ Balance
requiring minimum
interferences
BASIC SEPARATION TECHNIQUES
○​ Most pure
○​ Require maximum water
purity for accuracy and ★​ CENTRIFUGATION
precision ○​ Process in which centrifugal
○​ Use: flame photometry, AAS, force used to separate solid
blood gases and pH matter from a liquid
suspension
★​ TYPE 2 REAGENT WATER ○​ 3 variables: mass,speed and
○​ Used for hematology, radius
microbiology, immunology ○​ Speed of centrifuge is easily
and chemistry (MICH) checked using a tachometer
○​ Acceptable for preparation of (measure speed of
reagents and quality centrifuge) or a strobe light
materials (for the calibration of
centrifuge)
★​ TYPE 3 REAGENT WATER
○​ Utilized for preparation of
TYPES OF DESCRIPTION
reagents for urinalysis,
CENTRIFUGE
parasitology and histology
(UPAH)
Horizontal-hea -​ Cups holding the
○​ Used for washing glassware
d swinging tubes
bucket -​ Vertical position
NOTES TO REMEMBER centrifuge when at rest
➔​ Tests for water purity: microbiological -​ Horizontal position
content, pH, resistivity, ammonia, when revolves
ions and metals
➔​ Water may be distilled more than Fixed-angle -​ Cups are held in a
once head/angle-hea rigid position at a
➔​ Water can also be purified by d centrifuge fixed angle
ultrafiltration -​ More rapid than
➔​ Waters can be classified in terms of the horizontal
types
ultracentrifuge -​ High speed
centrifuge,
CLINICAL LABORATORY SUPPLIES separate layers of
different specific
gravity
★​ thermometers/ temperature
-​ Used to separate
★​ Laboratory vessels
lipoproteins
★​ Pipette
★​ Burette
Cytocentrifuge -​ Uses a very
★​ Syringes
high-torque and

CLIN CHEM 1 GWYN 3

 CLINICAL CHEMISTRY LECTURE
PRELIMS | 2ND SEMESTER
2-E | PROF LACIA

low inertia motor LABORATORY TESTING CYCLE

-​ Uses for blood,
urine, body fluid
★​ PRE-ANALYTICAL
and other spx that
○​ This is the phase that
can be spread on a
happens before the actual
slide
testing
○​ Patient preparation, patient
★​ FILTRATION identification and spx
○​ Utilizes filter papers to collection
separate solids from liquids ○​ End on centrifugation
○​ Liquid that passes through
the filter paper is called ★​ ANALYTICAL
filtrate ○​ Actual testing phase

★​ DIALYSIS ★​ POST-ANALYTICAL PHASE

○​ Separate macromolecules ○​ Encoding of results
from a solvent or smaller ○​ Happens after the actual
substance testing ​

PHLEBOTOMY AND SPECIMEN

CONSIDERATIONS

PATIENT PREPARATION

PRE-ANALYTICAL VARIABLES-FACTORS
CONTRIBUTING TO THE VARIATION OF
RESULTS:
★​ Exercise
★​ Fasting
○​ FBS 8-10hrs (to determine
blood glucose in the body)
/OGTT 8-10hrs (determine
how well our body
metabolizes sugar) and lipid
profile 10-12hrs
★​ Diet
★​ Tourniquet application
★​ Tobacco smoking
★​ Alcohol ingestion
★​ Stress
★​ Drugs
★​ Physiologic variations ​

CLIN CHEM 1 GWYN 4

 CLINICAL CHEMISTRY LECTURE
PRELIMS | 2ND SEMESTER
2-E | PROF LACIA

○​ Identified by asking the

attending nurse or relative
TYPES OF SAMPLES
★​ INFANTS AND CHILDREN
○​ A nurse/ relative may
★​ ARTERIAL BLOOD identify the patient; ID
○​ Oxygenated blood w/ a bracelet ​
bright red color
○​ Uses: blood gas analysis, ph ★​ OUTPATIENT/AMBULATORY
measurement ​ PATIENT
Primary sites: radial, ○​ Verbally ask their full names,
brachial and femoral arteries address or birth date
○​ Blood sample is collected
w/out a tourniquet GENERAL METHODS OF BLOOD
○​ Collected by a doctor. COLLECTION
Respiratory therapist ​

★​ WHOLE BLOOD (venous blood) ★​ ARTERIAL PUNCTURE

○​ Uses both the plasma and ★​ VENIPUNCTURE
cellular components ○​ Blood is obtained from a
○​ Requires an anticoagulant patient’s vein
○​ Spx for A1C test (HbA1C) ○​ The deoxygenated blood w/
determine blood glucose dark red color
control in the span of ○​ Sites: antecubital fossa, veins
2-3months on the wrist and dorsal
aspect of hands, veins on the
ankle
SPECIMEN COLLECTION AND
HANDLING
CAP ANTICOAGULAN INVERSIO
★​ Proper patient identification is the COLOR T/ADDITIVES NS
first step in sample collection ​
Yellow top SPS (sterile) 8-10x
★​ CONSCIOUS sodium
INPATIENTS/HOSPITALIZED polyanethol
PATIENTS sulfonate
○​ Verbally ask their full names
○​ Verify the name using the ID Light blue Trisodium citrate 3-4x
bracelet top
○​ 3-way ID (patient
identification, ID band, visual Red top w/wout clot 0 (glass)
comparison) activator or gel 5x (plastic)
separator
★​ SLEEPING PATIENTS
○​ Must be awakened before Green top Heparin (Li, Na, 8-10x
blood collection NH4)

★​ UNCONSCIOUS, MENTALLY lavender/ EDTA 8-10x

INCOMPETENTS purple top

CLIN CHEM 1 GWYN 5

 CLINICAL CHEMISTRY LECTURE
PRELIMS | 2ND SEMESTER
2-E | PROF LACIA

○​ Cardiovascular complications
Grey top Sodium fluoride 8-10x
○​ Dermatologic complications
and potassium
oxalate
★​ CAUSES OF HEMOLYSIS
(hemolyzed specimen)
○​ Using a needle that is too
NOTES TO REMEMBER
small
➔​ Tourniquet application (3-4 inches
○​ Pulling a syringe plunger
above the site, <1 minute)
back too fast
➔​ Cleanse site w/ friction using 70%
○​ Expelling the blood strongly
isopropyl alcohol
into a tube
➔​ Benzalkonium chloride (zephiran):
○​ Forcing the blood from a
used for ethanol testing
syringe into an evacuated
➔​ 70% alcohol followed by iodophor:
tube
most common form of skin cleansing
○​ Shaking or mixing the tubes
for blood culture
vigorously
➔​ Chlorhexidine gluconate:
○​ Performing blood collection
recommended for skin disinfectant by
before the alcohol has dried
CLSI for infants, children and acults
at the collection site
➔​ Traumatic draw as a result of vessel
wall in injury can cause increase CK,
★​ CAUSES OF HEMATOMA
myoglobin and potassium
○​ The vein is too fragile or too
small for the needle size
★​ Green - 21 gauge ○​ The needle penetrates all the
★​ Black - 22 gauge way through the vein
★​ Blue - 23 gauge ○​ The needle is partly inserted
★​ Purple - 24 gauge into the vein
★​ Orange - 25 gauge ○​ The needle is removed while
the tourniquet is still on
★​ SITES TO BE AVOIDED ○​ Excessive probing
○​ Intravenous lines in both ○​ Pressure is not adequately
arms applied after venipuncture
○​ Burned or scared areas
○​ Areas w/ hematoma ★​ SKIN PUNCTURE
○​ Thrombosed veins ○​ Fingerstick to obtain blood
○​ Edematous arms for routine lab analysis
○​ partial/radical mastectomy ○​ Useful in adults w/extreme
on one or both arms obesity, severe burns, and
○​ Arms w/arteriovenous (AV) thrombotic tendencies
shunt or fistula PREFERRED SITES
○​ Casts on arms ​ ○​ Lateral plantar heel surface
(newborn)
★​ COMPLICATIONS OF ○​ Palmar surfaces of the
VENIPUNCTURE fingers (3rd and 4th fingers)
○​ Vascular complications ○​ Plantar surface of the big toe
○​ Infections ○​ Earlobes (least site)
○​ Anemia
○​ Neurologic complications

CLIN CHEM 1 GWYN 6

 CLINICAL CHEMISTRY LECTURE
PRELIMS | 2ND SEMESTER
2-E | PROF LACIA

○​ hemolysis/lipemia
○​ Hemoconcentration
INTERFERING CONDITIONS IN THE
○​ Exposure to light/extreme
MEASUREMENT OF ANALYTES
temperatures
○​ Improperly timed
★​ HEMOLYSIS spx/delayed delivery to the
○​ Ruptured of RBCs laboratory
○​ Processing errors
★​ ICTERIC SAMPLE
○​ Increased bilirubin pigment
REASONS FOR SPX REJECTION
★​ LIPEMIA
○​ Increased lipids ○​ hemolysis/lipemia
○​ Clots in an anticoagulated
STORAGE OF SPECIMENS tube
○​ Non Fasting specimen
★​ Samples should be analyzed within ○​ Wrong blood collection tube
4hrs ○​ Short draws
★​ Serum or plasma must be stored at ○​ Improper transport (temp)
4’c to 6’c if analysis is to be delayed ○​ Discrepancies between
than 4hrs requisition and spx label
○​ Unlabeled or mislabeled
specimen
○​ Contaminated spx/leaking
OTHER BODY FLUIDS
container

★​ URINE
○​ Next most common sample
in clinical chemistry

★​ CEREBROSPINAL FLUID (CSF)

○​ Ultrafiltrate of plasma
★​ SYNOVIAL FLUID
★​ PLEURAL, PERICARDIAL,
PERITONEAL FLUID
★​ AMNIOTIC FLUID

TEN COMMON ERRORS IN

SPECIMEN COLLECTION

○​ Misidentification of px
○​ Mislabeling of spx
○​ Short draw/wrong
anticoagulant: blood ratio
○​ Mixing problem/clot
○​ Wrong tubes/wrong
anticoagulant

CLIN CHEM 1 GWYN 7