Southern Technical University
College of Health and Medical Technology/ Basra
Department of Medical Laboratory Technology
Clinical Biochemistry Subject
Fourth Class
Lecture No. Two
Specimens Collection
Lecturer
Abdullah Abbas Al-Rubaye
2023/2024
Fourth Class
Lecture No. Two Lec. Abdullah Abbas Al-Rubaye
Specimens Collection
Broad Objective
To be able to correctly collect specimen, put the specimen in the right container and
transport specimen to the laboratory correctly and at the right time.
Specimen
It is any material taken from the patient and sent to the laboratory for analysis.
Types of Specimens in clinical chemistry
• The biological fluids employed in the clinical biochemistry laboratory
include blood, urine, saliva, sputum, tissue and cells, cerebrospinal fluid,
peritoneal fluid, synovial fluid, pleural fluid, seminal fluid and stones.
• The process of specimen collection, handling and processing remains one of
the main causes of pre-analytical errors, in some studies they cause about
68% of laboratory errors.
Blood specimen
blood (whole blood or in the form of plasma or serum) is frequently used for
the investigations in the clinical biochemistry laboratory.
Sites of blood specimen collection
1- Skin punctures
Skin punctures are performed when small quantities of capillary blood are needed for
analysis or when the client has poor veins. Capillary puncture is also commonly
performed for blood glucose analysis. The common sites for capillary puncture are:
1- Fingertip is the inner aspect of palmer fingertip used most commonly in children
and adults.
2- Heel is the most common and minimally invasive method to draw capillary blood
from an infant for medical testing.
3- Earlobe is used when the patient is in shock or the extremities are edematous.
Fourth Class
Lecture No. Two Lec. Abdullah Abbas Al-Rubaye
2- Vein puncture
Wrap an elastic band (tourniquet) around patient's upper arm to stop the flow of
blood. This makes the veins below the band larger so it is easies to put a needle into
the vein.
Clean the needle sit with alcohol.
Put the needle into the vein.
Attach a vacutainer tube to the needle
Remove the band from patient's arm when enough blood is collected.
Apply a gauze pad or cotton ball over the needle site as the needle is removed.
Apply pressure to the site and then a bandage to fill it with blood.
Blood collection Tubes
The most widely used tubes for blood collection are evacuated tubes (Vacutainers)
• Negative pressure facilitates collection
• Easy to use
• Sterile
• Universally used color-coded rubber stoppers to denote tube type.
• Tubes can contain various anticoagulants for the collection of whole blood or
plasma.
• Tubes can have additives for specific tests (glucose, metals)
Fourth Class
Lecture No. Two Lec. Abdullah Abbas Al-Rubaye
Red-top blood tubes
• Red-top tubes contain no anticoagulants or preservatives.
• Red-top tubes are used for collecting serum.
• 10-15 minutes is required to allow blood to clot before centrifuging.
• Used for blood bank specimens, some chemistries.
Gold-top blood tubes
• Gold (and “tiger”) top tubes contain a gel that forms a physical barrier between the
serum and cells after centrifugation.
• No other additives are present.
• Gel barrier may affect some lab tests.
Fourth Class
Lecture No. Two Lec. Abdullah Abbas Al-Rubaye
Gray-top blood tubes
• Used for Glucose measurement.
• After blood collection, glucose concentration decreases significantly because of
cellular metabolism.
• Gray-top tubes contain either: Sodium fluoride and potassium oxalate, or
Sodium iodoacetate.
• Both preservatives stabilize glucose in plasma by inhibiting enzymes of the
glycolytic pathway:
▪ NaF/oxalate inhibits enolase.
▪ Iodoacetate inhibits glucose-3-phosphate dehydrogenase.
Green-top blood tubes
• Green-top blood tubes contain either the Na, K, or lithium (Li) salt of heparin.
Most widely used anticoagulant for chemistry tests.
• Should not be used for Na, K or Li measurement.
• Can affect the size and integrity of cellular blood components and not
recommended for hematology studies.
• Heparin accelerates the action of antithrombin III, which inhibits thrombin, so
blood does not clot (plasma).
• The advantage of plasma is that no time is wasted waiting for the specimen to
clot.
Fourth Class
Lecture No. Two Lec. Abdullah Abbas Al-Rubaye
Lavender-top blood tubes
• Lavender-top tubes contain the K salt of ethylenediaminetetraacetic acid
(EDTA), which chelates calcium (essential for clot formation) and inhibits
coagulation.
• Used for hematology, and some chemistries.
• Cannot be used for K or Ca tests.
Blue-top blood tubes
• Blue-top tubes contain sodium citrate, which chelates calcium and inhibits
coagulation.
• Used for coagulation studies because it is easily reversible.
Fourth Class
Lecture No. Two Lec. Abdullah Abbas Al-Rubaye
Brown and Royal Blue-top blood tubes
• Brown and Royal Blue top tubes are specially cleaned for trace metal studies
• Brown-top tubes are used for lead (Pb) analysis.
• Royal blue-top tubes are used for other trace element studies (acid washed).
Biochemical investigations
Biochemical investigations can be performed on 4 types of blood specimens - whole
blood, plasma, serum and red blood cells.
l. Whole blood (usually mixed with an anticoagulant) is used for the estimation of
glycated haemoglobin (HbA1c), carboxyhemoglobin, pH, glucose, urea, non-
protein nitrogen, pyruvate, lactate, ammonia etc.
2. Plasma obtained by centrifuging the whole blood collected with an
anticoagulant, is employed for the parameter- fibrinogen, glucose, bicarbonate,
chloride, ascorbic acid act.
3. Serum is the supernatant fluid that can be collected after centrifuging the clotted
blood. It is the most frequently used specimen in the clinical biochemistry
laboratory. The parameters estimated in serum include proteins, (albumin/
globulins), creatinine, bilirubin, cholesterol, uric acid, electrolytes (Na +, K+, Cl,
Ca¯), enzymes, vitamins and hormones.
4. Red blood cells are employed for the determination of glucose 6-phosphate
dehydrogenase, pyruvate kinase ect.
Fourth Class
Lecture No. Two Lec. Abdullah Abbas Al-Rubaye
Urine specimen
Urine specimen remains an important tool for clinical diagnosis. A correct urine
result is influenced by the collection method, timing and handling. The laboratory
test ordered determines the type of container to be used for collecting a specimen.
Types of urine collection
Laboratory urine specimens are classified by the type of collection conducted or by
the collection procedure used to obtain the specimen.
1- Random urine specimen
This is the specimen most commonly sent to the laboratory for analysis, primarily
because it is the easiest to obtain and is readily available. This specimen is usually
submitted for urinalysis and microscopy analysis. Although there are no specific
guidelines on how to collect the specimen, avoiding the introduction of
contaminants into the specimen is recommended, by advising the patient not to
touch the inside of the cup or cup lid.
2- First morning urine specimen
This is the specimen of choice for urinalysis and microscopic analysis, since the
urine is generally more concentrated due to the length of time urine is allowed to
remain in the bladder and therefore, contains relatively higher levels of cellular
elements and analytes such as protein, if present. Also called an 8-hour urine
specimen, the first morning specimen is collected when the patient first wakes up in
the morning, having emptied the bladder before going to sleep.
NOTE: any urine that is voided from the bladder during the eight hours collection
period is pooled and refrigerated, so that a true 8hour specimen is obtained.
3- Midstream clean catch urine specimen
This is the preferred type of specimen for culture and sensitivity testing because of
the reduced incidence of cellular and microbial contamination. The patient should
then void the first portion of the urine stream into the toilet. These first steps
significantly reduce the opportunities for contaminants to enter into the urine
Fourth Class
Lecture No. Two Lec. Abdullah Abbas Al-Rubaye
stream. The urine midstream is then collected into a clean container, (any excess
urine should be voided into the toilet). This method of collection can be conducted
at any time of day.
Sampling errors
There are a number of potential errors which may contribute to the success or
failure of the laboratory to provide the correct answers to the clinician's question.
Some of these problems arise when a clinician first obtains specimens from the
patient.
1- Blood sampling technique
Difficulty in obtaining a blood specimen may lead to haemolysis with consequent
release of potassium and other red cells constituents. results for these will be falsely
elevated.
2- Prolonged stasis during vein puncture
Plasma water diffuses into the interstitial space and the serum or plasma sample
obtained will be concentrated. Proteins and protein-bound components of plasma
such as calcium or thyroxine will be falsely elevated.
3-Insufficient specimen
Each biochemical analysis requires a certain volume of specimen to enable the
test to be carried out it may prove to the impossible for the laboratory to measure
everything requested on a small volume specimen.
4- Errors in timing
The biggest source of error in the measurement of any analyte in a 24-hour urine
specimen is in the collection of an accurately timed volume of urine.
5- Incorrect specimen container
For many analyses the blood must be collected into a container with anticoagulant
and preservative. For example, samples for glucose should be collected into a
Fourth Class
Lecture No. Two Lec. Abdullah Abbas Al-Rubaye
special container containing fluoride which inhibits glycolysis; otherwise, the time
taken to deliver the sample to the laboratory can affect the result.
If a sample is collected into the wrong container, it should never be decanted into
another type of tube. for example, blood which has been exposed even briefly to
EDTA (an anticoagulant used in sample containers for lipids) will have a markedly
reduced calcium concentration, approaching zero.
6- Inappropriate sampling site
Blood samples should not be taken 'down-stream' from an intravenous drip. It is not
unheard of for the laboratory to receive a blood glucose request on a specimen taken
from an intravenous drip. It is not unheard of for the laboratory to receive a blood
glucose request on a specimen taken from the same arm into which 5% glucose is
being infused. Usually, the results are biochemically incredible but it is just possible
that they may be acted upon, with disastrous consequences for the patient.
Case study: a blood sample had been taken from 44 years; old
woman the results were as follows:
-Sodium 144 mmol/L (135-145 mmol/L).
-Urea 4.1 mmol/L (2.5-7.0 mmol/L).
-Creatinine 78 mmol/L (70-110 mmol/L).
-Calcium 0.5 mmol/L (2.5-2.55 mmol/L).
Repeat blood sampling on the same day gave the following results.
-Sodium 139 mmol/L (135-145 mmol/L).
-Urea 4.2 mmol/L (2.5-7.0 mmol/L).
-Creatinine 78 mmol/L (70-110 mmol/L).
-Calcium 2.43 mmol/L (2.5-2.55 mmol/L).
Discuss these results?