INTRODUCTION

TO CLINICAL
BIOCHEMISTRY
LAB
OBJECTIVE

Students are expected to:
Explain types of biochemistry test with examples.
Describe types of specimen available.
Understanding sources of sampling error.
Explain the factors contributing variation of results.
Understanding the importance of the Clinical
biochemistry lab.
Expecting General Risk
Describe Clinical Biochemistry Process
Elaborate Quality Assurance (QA) & Quality Control
(QC)
Types of Biochemistry Test
Core Test Special Test Emergency Test
- test which are commonly
requested that are value in
many patients on a frequent
basis.
- test which are needed
in diagnosing rare
disease.
-test which are requested
urgently.
- sodium, potassium, chloride
& bicarbonate
- urea & creatinine
- calcium & phosphate
- bilirubin & alkaline
phosphatase
- alanine aminotransferase
- creatin kinase
- acid-base imbalance
- glucose
- total protein & albumin
- hormones
- specific proteins
- trace elements
- vitamins
- drugs
- lipids & lipoprotein
- DNA analysis

- urea & electrolyte
- blood gases
- amylase
- glucose
- salicylate
- paracetamol
- calcium
Type of Specimen
Types of Specimen
Blood
Serum
- blood which is collected into plain tube that allowed to
clot in room temp.
- recommended for biochemical analysis
Plasma
- used when the analyte questioned is unstable
- obtained when blood collected in an anticoagulant tube
- when centrifuged, the supernatant is called plasma
Urine
Cerebrospinal fluid
Others?
SPECIMEN COLLECTION
The test request
The specimen for analysis must be collected and
transported to the laboratory according to a specified
procedure if the data are to be of clinical value. This
procedure begins with the clinician making a test
request, either on a computer terminal or on paper. The
completed request should include:
patient's name, sex and date of birth
hospital or other identification number
ward/clinic/address
name of requesting doctor (telephone/pager number for
urgent requests)
clinical diagnosis/problem
test(s) requested
type of specimen
date and time of sampling
relevant treatment (e.g. drugs).

It is essential that sufficient information be provided
to identify the patient. In practice, vital information is
often omitted and this may either cause delay in
analysis and reporting or make it impossible to
interpret the results.

Sampling Error
Blood sampling technique
-improper technique lead to hemolysis and give false
positive result with consequent increase of potassium
and red cell constituents.

Prolong stasis during venipuncture
- stasis causes plasma water diffuse into interstitial
space and serum or plasma sample obtained will be
concentrated.
- protein and protein bound components of plasma such
as calcium or thyroxine will be falsely elevated

Insufficient specimen
- certain volume of specimen is required to enable the
test to carried out

Errors in timing
- collection of an accurately time volume of urine such as
24 hrs urine

Incorrect specimen storage
- blood sample stored overnight before being sent to lab
will show falsely high K+, phosphate and red cell
enzymes such as LDH because of leakage into the
extracellular fluid (ECF) from the cells.

Inappropriate sampling site
- blood taken downstream from IV drip may affected by
the drip content

Incorrect specimen container
-anticoagulant/ preservative. Eg: glucose in fluoride tube
which inhibits glycolysis.
-never decanted into another tube.
Variation Result
Variation Result
Due to biological factors:
Gender-muscle mass
Age- increase in ALP
Diet-fasting,malnutrition,alcoholic
Pregnancy- insulin resistance
Time the sample taken- cortisol
Stress and anxiety
Medical and drug history
Importance of Biochemistry Lab
Screening detection of subclinical disease
Diagnosis confirmation or rejection of
clinical diagnosis
Prognosis information regarding the likely
outcome of disease
Monitoring monitoring progression or
response to treatment
General Risk in Lab Test
Determination of the right test
- to decide the most sensitive, specific, accurate and
precise with reasonable price

Sample collection procedure
- collected with right manner. Eg: avoid hemolysis during
blood collection
- collected from right source. Eg: CSF to determine
meningitis

Sample transportation
- use the right tubes. Eg: tubes with sodium citrate for
glucose

Sample storage
- serum separated from blood cells before storage
- urine should not be left more than 2 hours unless
refrigerated

Human error
- unskilled lab personnel

Interpretation of result
- use the right reference range
Clinical Biochemistry Process
Clinical questions
Request form & clinical data
Specimen collection
Transit sample to lab
Reception at lab and ID
Analysis
QC
Collation
Result interpretation
Reporting to doctor
Biochemistry answer
Quality Assurance (QA) & Quality
Control (QC)
Pre Analytical Quality Assurance
Analytical Quality Assurance
Post Analytical Quality Assurance

Quality Control
Quality Assurance (QA)
QA - all activities that ensure the validity of
chemical testing and analysis and all relevant
technical support
Lab personnel required to follow QA plans and
SOP
These documents and staff vigilance help
ensure the analytical data and other test results
will be acceptable as the bases for making
significant decision.
Pre Analytical Error
Patient preparation
- nutrition status, recent meal, alcohol, drugs, smoking,
exercise, stress

Sample collection and handling
- right specimen collection tubes usage
- sample match with anticoagulant or additive
- specimen transportation and separation
- avoid contamination
-effects of infusion
- hemolysis
Storage
- to know the characteristics of each analyte and store
with right manner in order to preserve the analyte
amount
- Evaporated analyte to store with tightly capped. Eg:
electrolytes
- Light sensitive analyte to be stored in dark place. Eg:
bilirubin
- Temperature sensitive analyte to be refrigerated. Eg:
Lactate dehydrogenase

Analytical QA
To decide on the best instrumentation or
analyzer
To choose the right and suitable reagents
Post Analytical QA
System to record all patients profile to avoid
losing data
System to report the reliable lab data to the
doctors in specific time given
Quality Control (QC)
Refers to the measures that must be included
during each test run to verify that the test is
working properly.
Aims to recognize and minimize analytical
errors.
Management requirement
ISO 9000
-certification:
-documented quality system is in place
-Auditors are skilled in Quality Assurance but
may not have the expertise in service.
ISO 15189
-more specific for lab operation
-technically competant, have laboratory
experience, skilled in lab management, QA
technique
ANALYSIS OF BLOOD
BIOCHEMISTRY
Learning objectives:
Describe blood composition
Differentiate types of blood sample and its
source
Explain causes lead to hemolysis and how to
avoid hemolysis
Identify the blood collection tubes and the
usage of the tubes
Blood Composition
Cells
Erythrocytes
Leucocytes
Platelet

Plasma vs serum?? - nutrient, metabolism
product
Plasma vs serum
Type of Blood Sampling
Venous blood
Capillary blood
Drawn from finger tip and heel
Mixed between arterial and venous blood
Arterial blood- for blood gases test

Blood collection on babies heel punctures

Hemolysis
Due to - high speed centrifugation
- high pressure during draw the blood

Hemolysis avoided by:
Use dry and clean needle and syringe
Apply minimal constriction on arm
Let blood flow slowly into syringe
Use dry blood tube
Mix well blood with anticoagulant
Low velocity centrifugation

Blood Tubes

Type of Blood Collection Tube
Stopper
Colour
Additive

Resulting
Sample
Additive
Action
Assay Comments
Red None Serum None Enzymes,
electrolytes,
iron, lipids,
drugs level
Some tube have
silicone coating
to accelerate
clotting
Red/
Grey
Inert
polymer
barrier
Serum Cleaner
separation of
serum and
cell
Same as red
stopper
Require for CO2
analysis
Orange Thrombin Serum Accelerate
clotting
Stat
chemistry
Emergency
purposes
Grey Iodoacetate Serum Inhibit
glycolysis
Glucose,
lactose
No interference
with enzymatic
glucose/BUN
Stopper
Colour
Additive Resulting
Sample
Additive
Action
Assay Comments
Grey Na fluoride &
K oxalate
Plasma Inhibit
glycolysis
Glucose,
lactose
Not for Na & K
analysis
Green Na/Li/NH3
Heparin salts
Plasma Enhance
antithrombin
III
Cortico
steroids,
electrolytes
Antithrombin
III inactivates
clotting factors
Brown Na heparin Plasma Enhance
antithrombin
III
Lead Tube has only
minute
quantity of
lead
Royal
blue
None Serum None Trace
elements
Tube has only
minute
quantity of
trace
elements
Lavender EDTA Plasma Binds Ca CEA,
hematology
test
Ca is needed
for clotting
Blue Na citrate Plasma Binds Ca Coagulation
test
Ca is needed
for clotting
TUTORIAL
1. Define core test, specialized test and emergency test.
Give 2 examples of each.
2. List out types of specimen should be received in a
clinical biochemistry lab.
3. Discuss on how error could happen during sampling
procedures.
4. Explain the significant of Quality Assurance practices.
5. Elaborate preanalytical, analytical and post analytical
quality assurance in order to produce a reliable result.
NEXT CLASS
ACTIVITY 1

Student need to form group of 4 and number
each group member. Propose a procedure on
drawing blood sample and decide on the best
tube used for these situations:
a) To monitor blood glucose level
b) To investigate bleeding tendency
c) To determine cardiac enzyme level in
blood
Justify your answer.