LESSON 3: SPECIMEN COLLECTION AND PATIENT PREPARATION

QUESTIONS FOR RESEARCH:

1. What types of specimens are useful for clinical chemistry analysis and what specimen types are
required for specific chemistry tests? How are these specimens collected?

Clinical chemistry makes use of chemical procedures to gauge chemical component

concentration levels in tissues and bodily fluids. Blood and urine are among the most usual
specimens used in clinical chemistry.

To obtain and assess about every type of chemical component in blood or urine, there are lots
of different testing methods. Blood glucose, electrolytes, enzymes, hormones, lipids (fats), other
metabolic substances, and proteins are examples of its components.

Some of the most typical laboratory examinations include the ones listed below:
 A urine analysis is a lab test which then looks for various chemicals and cells, such as
red or white blood cells, infections, or an excessive amount of protein, in urine. Urine is
separated into its component parts in order to detect the presence of drugs, blood,
protein, and other substances. Blood within that urine, or hematuria, can be brought on
by a benign (noncancerous) situation, but it may also be an indication of an infection or
other problem. A high level of protein in the urine, or proteinuria, may indicate kidney or
cardiovascular problems.
 Blood tests are used to examine genetics, various blood chemistries, inflammation
markers, and cell counts.
 Tumor markers are substances that are either produced by the body in response to
cancer cells or substances that are released by cancer cells into the blood or urine.

2. How do different anticoagulants affect chemistry tests? Which anticoagulants are best for specific
chemistry assays?

Anticoagulants and additives in plasma samples can adversely affect sample stability, protein
binding to the target analyte, and the analytical characteristics of the assay. Liquid
anticoagulants may also cause the sample to be improperly diluted.

Due to their special characteristics, some anticoagulants are used for particular assays.
Examples include fluoride, which blocks the activity of phosphorylating enzymes and yields
accurate blood sugar estimates, and citrate, which is used at particular concentrations for
coagulation studies.

 ETHYLENE DIAMMINE TETRAACETIC ACID, or EDTA

In laboratories, it is a chemical anticoagulant that is frequently used. The chelating
agent EDTA is colorless, water soluble, and has the capacity to bind to metal ions like
calcium. Its dosage ranges from 1.25 to 1.75 milligrams per milliliter of blood. Blood
drawn during an EDT procedure can be used for TLC, PS preparation, hemoglobin
estimation, and differential leukocyte count (DLC).
 TRI-SODIUM CITRATE
In the early twentieth century, this was first utilized as an anticoagulant in blood
transfusion. Its application in the blood collection tubes for ESR and coagulation

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 studies, and even for the preservation of blood in blood banks, is all still useful and
relevant to this day.
 OXALATES
They can be utilized as single oxalates, such as sodium oxalate, potassium oxalate, or
ammonium oxalate, however they are frequently used in double oxalate due to the fact
that when it is used alone, potassium oxalate at a concentration of 2 mg/ml of blood
shrinks red blood cells (RBCs), whereas ammonium oxalate at a concentration of 2
mg/ml may end up causing RBC swelling.
 SODIUM FLUORIDE
It is the anticoagulant of choice for the estimation of blood sugar.
 HEPARIN, A BIOLOGICAL / NATURAL ANTICOAGULANT
Heparin is a naturally occurring anticoagulant that cannot be made in a laboratory. It's
gone and got from a leech. It works well as an anticoagulant and maintains the shape of
red blood cells (RBCs).

3. What causes specimen hemolysis, icteria, and lipemia? Which chemistry analytes are most affected
by these?

Hemolysis can occur intravenously or extravenously. Intravascular hemolysis is extremely

uncommon and typically brought on by hemolytic anemia or a blood transfusion reaction.
Extravascular hemolysis is quite common and happens when specimens are handled,
transported, and stored improperly during phlebotomy procedures.

Increased Decreased
Potassium (K+) Troponin T
SGOT/AST Haptoglobin
SGPT/ALT Bilirubin
Lactate Dehydrogenase (LDH) Amylase
Creatine Kinase (CK) Bicarbonate (HCO3–)
Iron
Phosphate (PO4—)
Total Protein
Albumin
Magnesium (Mg++)
Calcium (Ca++)
Alkaline Phosphatase (ALP)

Irregular excretion or increased bilirubin production both contribute to icterus. For instance:
biliary tract obstruction, liver disorders, hemolytic anemia, etc.

Increased Decreased
Magnesium (Mg++) Cholesterol
Triglyceride
Creatinine
Bile Acids
Lipase
Total Protein

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 Uric Acid
GGT

The condition known as lipemia is characterized by an elevated level of triglyceride-rich

lipoproteins in the blood, which gives serum or plasma a cloudy or turbid appearance. The size
of lipoproteins varies, so not all of them equally contribute to turbidity. Chylomicrons, the largest
particles, have the greatest potential to muddy the sample.

Increased Decreased
Bile Acids Sodium (Na+)
Direct Bilirubin Potassium (K+)
TIBC Chloride (Cl–)
Magnesium (Mg++) Bicarbonate (HCO3–)
Lactate Dehydrogenase (LDH)

4. What is "order of draw" and how does this affect chemistry analysis? What other factors of specimen
collection can have an adverse effect on chemistry analytes?

The recommended order for drawing blood samples is referred to as the "order of draw" in effort
to reduce or nullify intervention in testing caused by carryover of additives in tubes. The
additives in one tube may be transferred to another if the order of draw is not generally followed,
which could lead to contaminated samples and inaccurate results. An incorrect diagnosis could
result from inconsistent data, which would keep patients from getting the right treatment.

Analytical changes, which result from mistakes in test procedures and equipment, as well as the
timing of an investigation in the case of illness, can also negatively affect chemistry analytes.
The importance of an investigation depends on how close the time of sample collection was to
the stage of the illness. The stage of the illness can also be impacted by the choice of
investigation. If the sample is drawn too soon after exposure, for example, the serology test may
be negative for syphilis; biological changes—there are some changes in lab test results that we
can anticipate due to immutable factors. These include biological rhythms, aging, and
pregnancy-related physiological changes.

6. Why are preanalytical variables important to be aware of when performing clinical chemistry testing?
Identify and list the pre-analytical, analytical and post-analytical variables that can adversely affect
laboratory analyses and effects on test result.

The pre-analytic stage is crucial to the overall quality of the laboratory. The pre-analytic phase
includes all of the steps taken during the collection, handling, and processing of a patient's
sample of blood or body fluid in order to prepare it for analysis. This phase also includes a

variety of other factors that can influence the outcome for that patient. The pre-analytic phase
includes factors such as pregnancy and menstruation as well as physiological variables like
lifestyle, age, and sex. Endogenous factors, like medications or circulating antibodies, may
interact with a particular technique to produce erroneous analytical results. Variables in the pre-
analytic phase have an impact on a variety of laboratory disciplines.

The pre-analytical stage is the first of these steps, and it can start even before the specimen is
brought to the lab. Thus, strict protocols must be in place to lower the possibility of mistakes
being made when handling and identifying specimens. Sample type, sample timing, sample
handling, and sample identification are pre-analytical different factors that may have an impact
on test outcomes.

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 Analytical phase: The analytical phase is the second stage. When the samples have entered the
laboratory, this starts. The laboratory testing and diagnostic procedures make up this phase.
The analytical phase may involve both human and mechanical errors. During the analytical
phase of laboratory testing, the following mistakes may occur:

 Problems with performance specifications' quality assurance and verification

 Errors in collaboration
 Reagent blunders
 Mistakes caused by manual pipetting
 Interference caused by unknown antibodies
 Mistakes in mathematics
 Clinician mistakes in processing and preparation
 Errors brought on by staff fatigue
 Analytical quality is still a serious problem despite automation. For instance, problems
can occur as a result of improper sample preparation or the unintentional presence of
substances that can affect the precision, accuracy, and sensitivity of results.

Post-analytical Phase: The disseminating of the ultimate result is part of the post-analytical
phase. The necessary person receives the results promptly. When results are manually entered
and stored, data entry mistakes may occur. Data transmission can also result in post-analytical
errors. Verbal information can occasionally be misheard or incorrectly transmitted. Among the
other frequent errors with post-analytical test results are:

 Erroneous calculations
 Delayed results turnaround time
 Results delivered to the incorrect person
 No results are provided.

7. Outline the general steps for processing blood samples. Elucidate the complications of venipuncture
and skin puncture.

It's crucial to collect, handle, and process samples properly. Errors made during sample
collection and transportation account for the vast majority of incorrect results. Because of this,
it's crucial that the right procedures for patient preparation, sample preparation, sample
collection, and sample transport are followed.
 Identify the patient; two active identification methods are needed.
 Assure the patient that only the bare minimum of blood will be drawn to conduct the
test.
 Check to see if any dietary or time restrictions have been adhered to.
 Order of Drawing.

If venipuncture is performed improperly, there may be complications including increased risk of

accidental needlesticks, the possibility that hemoconcentration will affect test results, an
increased risk of hematoma formation, a need for greater patient stability, the possibility that
multiple venipunctures will cause iatrogenic anemia, and difficulty identifying infant nerve
damage.

An improper skin puncture could result in consequences such hemolysis, specimen clotting,
increased patient discomfort, decreased blood yields, the possibility of tissue fluid
contamination, and prolonged procedure times.

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8. What are the aspects of specimen handling and how can mishandling of samples affect clinical
chemistry values?

The suitable technique is always to handle specimens in accordance with evidence-based

guidelines for a specific research design. The exact assays that will be used, nevertheless,
cannot always be predicted. In those circumstances, samples should be taken in accordance
with procedures that are most likely to ensure the efficient preservation of the biospecimen in a
condition that most closely resembles that of the patient before it was removed.

It is essential to have the proper cryovials, vacutainers, and other containers on hand at the
collecting location, and those containers should be capable of supporting the samples' expected
downstream applications. Anticoagulants and other stabilizers of biological samples may be
necessary for some downstream uses but harmful for others. Additionally, some items may have
expiration dates that specify a limited time frame for appropriate use (e.g., vacutainers, tissue
culture media, and antibiotics). To ensure that supplies and reagents are used within the allotted
time range, it is imperative to keep track of expiration dates.

Presumably, the following are the effects of specimen handling errors on care delivery:

 Failure to give a patient the right care at the right time due to insufficiently precise test
results associated to the right patient.
 Wrongly treating a patient based on a test result that wasn't actually from that patient.

REFERENCES/SOURCES:

Indicate the learning materials you used in accomplishing this self-directed activity.

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