Sampling

Assistant instructor. Jinan Hameed

Source by:-Practical clinical biochemistry
• Specimen Collection

 What should we know about collection Samples ?

 How Care during and after Blood Sample Collection?
Blood Sample.
Types of Anticoagulants .
Urine Specimen .
Stool Specimen .
Cerebrospinal Fluid Sample (CSF) .
Complete specimen collection and handling requirements
are provided in the alphabetical test listing of the online
test catalog like:

specimen type
minimum volume
storage temperature, and other special handling notes
 Storage temperature

– specified as room temperature (18° to 22°C)

– refrigerated at (4° to 7°C)
– frozen at (-20°C or colder).
 Sample Collection

 The good way of sample collection and handling decided the correct and
incorrect results and the degree of errors. So, some results of some
patients do not related to the condition of patients, but appear due to
wrong sample collection.
 Frozen sample should not be submitted in a glass container, it should be
submitted in a plastic container.
For Collection of Sample we should
know the following:
– 1. Sample collection depends on the test requested and the method of analysis
as manual spectrophotometric method or automated analyzer method.
– 2. Use of preservative ex: sodium florid to prevent glycolysis in RBCs.
– 3. Should do sample labeling.
– 4. Patient previous diet as some tests need dietary preparation of the patient.
– 5. Patient treatment or self-medication and its effects on the measured analytes.
– 6. Time of the day for sampling is very important, as the concentration of some
analyte may be changed during the day (ex: cortisol and triglyceride levels the
early morning sample is preferred).
 Care during Blood Sample Collection

– a. The skin antiseptics should not interfere with the measured

analyte.
– b. Choosing the right vein for blood sampling.
– c. Fasting sample should be overnight (12 hours) is the best for
correct result.
– d. Patient should be at rest and lying for at least 20 minutes before
getting blood sample. It was found that there are 13% differences in
concentration of analyte between the sampling in case of lying and
standing due to the distribution of fluid in the extracellular space.
e. Prevent venous stasis, which will increase the analyte concentration
as in case of plasma protein, calcium and others. This occurs due to the
use of tourniquet for a long time before getting blood sample.

– f. Do not take sample from infusion arms.

– g. Prevent blood hemolysis which may be arises from the using of too large or too
small needle, moisture in the syringe, vigorous mixing of the blood, and rapid
expansion of the blood into the tube or due to the separation process.
Hemolysis may lead to mix the intracellular with the extracellular components.
Hemoglobin may directly interfere with a chemical determination by inhibiting an
enzyme such as lipase, by interfering with a reaction as that of diazo with bilirubin,
or by yielding a significant color and thus interfere with colorimetric analysis.
Drawing of blood in a color-coded vacuum tube is one of the important alphabetical things of
the specimen handling requirement, as shown in figure(1). Blood drawn from a patient by
syringe and transfer to clean dry tube after removal of the needle to prevent break down of
RBCs
Care After Blood Collection

– This includes the way of releasing blood from the

syringe, cooling and freezing,
– using of preservative and anticoagulant.
– Chemical investigations mainly run on the
specimens.
 Blood Sample
 Blood is a suspension of cells in a protein-salt matrix.

 The non-cellular portion of blood contains series of proteins some of which are
involved in the coagulation process; this fluid is called plasma. While the fluid
that is separated from the clotted material is called serum.
 Blood used for biochemical analysis is collected from the veins, arteries or
capillaries
 For most testing venous blood is utilized because of easiness of collection, but for
limited number of analyte such as blood gases and lactate there is a significant
difference arise between arterial and venous blood samples, in this conditions
arterial blood samples are preferred.
 How can you prepared the serum and plasma
sample?
 For serum preparation, allow the blood to clot at least for fifteen minutes,
as serum does not contain fibrinogen and clot formed without using anti-
coagulant and separated by centrifugation.
 Plasma contains fibrinogen so thoroughly mix the blood with anticoagulant
to stop clot formation, and gently invert the tube five to ten times and
prepared by centrifugation.
Blood clot formation
Chemical substances
that prevent or
reduce coagulation
of blood ,prolonging
the clotting time.
Types of Anticoagulants
Different types of anticoagulants are used in the medical laboratory tests depending on
the analyte measured. They are differing in their way of preventing coagulation

– Heparin: α-mucoitin polysulphuric acid inhibits formation of thrombin

from prothrombin. It is usually available as Na, K, NH4 and Li salts.
Oxalate and Citrate: as sodium oxalate, potassium oxalate (the most soluble
one), ammonium oxalate and lithium oxalate. Oxalate acts by precipitating
the calcium.
Ca++ precipitate

while sodium citrate converts calcium into non-ionized form.

Sodium Fluoride: It is also used as preservative for glucose
determination by inhibiting red cell metabolism through inhibiting
enzyme enolase that is involved in glycolysis
Ethylenediamine Tetra Acetic Acid (EDTA): It chelates calcium ions which are
essential for clotting mechanism. It is usually used as dipotassium and dilithium
salts.
 Urine Specimen

– Single fresh urine sample is used for general urine Stool Specimen
examination
– for most qualitative tests. For quantitative work, – Fresh stool specimens are
24 hour specimen collection is best employed at
important for general stool
which the patient first empties the bladder and
discards the urine, thereafter, all specimens passed examination
during the day and night are saved, added to them the – microscopic diagnosis of
sample of the following morning. different parasites, stool culture
 The sample is collected in a clean covered container and sensitivity tests. It can be
and kept in a cool place preferably in the refrigerator. used for occult blood test
It is necessary to prevent the effect of bacteria by
adding proper preservative such as hydrochloric
acid, chloroform or formalin to the urine.
Cup urine
 Cerebrospinal Fluid (CSF) Sample

 Cerebrospinal fluid is
formed by secretion from the
cells of choroid plexuses,
vascular structure lying
within the ventricles of the
brain and reabsorbed into
the blood stream by the
arachnoid villi.
 Normal &abnormal composition of CSF
appearance Clear, colorless Turbidity Meningitis & after hemorrhage
Excess of WBC(pus cells)

Lymphocytes 0-4*106 /L Increase in total protein All types meningitis

due to the increase in cell content and the
consequence inflammatory reaction
Total volume 125-150ml
clotting when there is an excess of due to high total protein content, also high protein
Specific gravity 1.005 fibrinogen in the specimen concentration arises from blockage of spinal canal
leading to stasis and fluid reabsorption

Total protein 0.15-0.45 gm/L

• infection like pyogenic and tuberculosis meningitis this lead to decrease in the glucose
Total globulins 6-15 mg/dl level in the CSF.
albumin 10-30 mg/dl • In viral meningitis the glucose level is normal
Glucose 40-75 mg/dl • In hyperglycemia, glucose level is elevated in the CSF but remains lower than glucose
of the blood
2.2-4.2 mmol
Thank you for
your attention