Key Takeaways

  • Centrifugation separates blood into its components — serum (from clotted blood) or plasma (from anticoagulated blood) is separated from cells
  • Serum separator tubes (SST/gold top) must clot for 30 minutes at room temperature before centrifugation; plain red tops require 60 minutes
  • Aliquoting is the process of transferring separated serum or plasma to a secondary tube for testing or storage
  • Common specimen rejection criteria include: hemolysis, lipemia, clotted anticoagulated specimens, wrong tube type, insufficient volume, and expired tubes
  • Quality assurance (QA) is a comprehensive program ensuring the entire testing process produces reliable results — includes preanalytical, analytical, and postanalytical phases
  • Delta checks compare a patient's current test results with their previous results to identify potential errors or significant clinical changes
  • The most common preanalytical errors are: patient misidentification, hemolyzed specimens, clotted anticoagulated specimens, and incorrect tube selection
  • Lab requisition forms must include: patient name, DOB/MRN, ordering physician, tests requested, date/time ordered, diagnosis/ICD code, and specimen source
Last updated: February 2026

Specimen Processing & Quality Assurance

Specimen processing is the bridge between collection and laboratory analysis. Proper processing techniques ensure the integrity of the specimen and the accuracy of test results. Quality assurance programs provide systematic oversight of the entire testing process.


Centrifugation

Centrifugation uses centrifugal force to separate blood into its components based on density. The heavier cellular components (RBCs, WBCs, platelets) settle to the bottom, while the lighter liquid component (serum or plasma) rises to the top.

Serum vs. Plasma

TypeSourceTubeContains Fibrinogen?Clotting Required?
SerumClotted bloodRed top, Gold SSTNo (consumed during clotting)Yes — must clot before centrifuging
PlasmaAnticoagulated bloodGreen (heparin), Lavender (EDTA), Light blue (citrate)Yes (fibrinogen preserved)No — anticoagulant prevents clotting

Clotting Times Before Centrifugation

Tube TypeClotting TimeReason
Gold SST (clot activator + gel)30 minutes at room temperatureClot activator accelerates clotting
Red top (no additive)60 minutes at room temperatureNo clot activator; natural clotting process takes longer
Anticoagulated tubes (green, lavender, light blue)No clotting time — centrifuge immediately if neededAnticoagulant prevents clotting

Centrifugation Guidelines

  • Standard speed: 1,000-2,000 x g (relative centrifugal force) for 10-15 minutes
  • Ensure the centrifuge is properly balanced (equal tube weights on opposite sides)
  • Never open the centrifuge lid while it is spinning
  • After centrifugation, the gel separator in SST tubes forms a barrier between serum and cells

Aliquoting

Aliquoting is the process of transferring separated serum or plasma from the original tube to a secondary (aliquot) tube:

  • Use a disposable transfer pipette — never pour from one tube to another
  • Label the aliquot tube with the same patient information as the original
  • Aliquoting is performed when:
    • Multiple departments need the same specimen
    • The specimen must be stored or frozen for later testing
    • A send-out laboratory requires a specific container
  • Avoid transferring any cells or gel into the aliquot tube

Serum vs. Plasma — Understanding the Difference

This is a commonly tested concept on the NHA CPT exam:

Serum

  • Produced from clotted blood (red top, gold SST)
  • Blood is allowed to clot, then centrifuged
  • Does NOT contain fibrinogen or clotting factors (they were consumed during clotting)
  • Appears as a clear, straw-yellow liquid above the clot
  • Used for most chemistry tests, serology, and immunology

Plasma

  • Produced from anticoagulated blood (green, lavender, light blue)
  • Blood is prevented from clotting by the anticoagulant
  • DOES contain fibrinogen and clotting factors
  • Appears as a clear, straw-yellow liquid above the packed cells
  • Used for coagulation tests, stat chemistry, some immunology tests

Key Differences at a Glance

FeatureSerumPlasma
ClottingBlood must clot firstClotting is prevented
FibrinogenAbsent (consumed)Present
Tube typesRed, gold SSTGreen, lavender, light blue
Processing timeLonger (30-60 min clotting + centrifuge)Shorter (centrifuge immediately)
Gel separatorGold SSTGreen/gray PST (plasma separator tube)
Common testsBMP, CMP, lipid panel, thyroidPT/INR, PTT, stat electrolytes, CBC plasma

Specimen Integrity Indicators

Modern laboratory analyzers can detect specimen quality issues automatically:

IndicatorAbbreviationAppearanceCause
HemolysisHPink to red serum/plasmaRBC destruction
IcterusIDeep yellow/orange serum/plasmaElevated bilirubin (jaundice)
LipemiaLMilky/turbid serum/plasmaElevated triglycerides (non-fasting)

These indicators (collectively called the HIL index) are reported with test results when they may affect accuracy. The laboratory may request a recollection if interference is significant.


Specimen Rejection Criteria

Laboratories reject specimens that do not meet quality standards. Common rejection reasons:

Reason for RejectionDescriptionPrevention
HemolysisPink/red serum or plasma (ruptured RBCs) — #1 cause of rejectionProper technique: correct gauge, don't shake, allow alcohol to dry
LipemiaMilky/turbid serum (high fat content)Ensure proper fasting; note non-fasting on label
Clotted anticoagulated specimenClots in EDTA, citrate, or heparin tubesMix tubes immediately by inversion; check for proper fill
Wrong tube typeTest ordered requires a different tube than what was collectedVerify order of draw and tube selection before drawing
Insufficient volumeNot enough blood for the test (especially critical for citrate tubes)Ensure proper tube fill; redraw if underfilled
Expired tubesTubes past their expiration date may have degraded vacuum or additivesCheck expiration dates before use
Mislabeled or unlabeledCannot confirm patient identityLabel at bedside; use two identifiers
Improper transportWrong temperature, excessive delay, broken tubeFollow transport protocols for each specimen type
QNS (Quantity Not Sufficient)Sample volume too low to perform the requested testCollect adequate volume; check tube fill

Laboratory Test Panels

Phlebotomists should be familiar with common test panels (groups of tests ordered together):

Basic Metabolic Panel (BMP) — 8 Tests

TestTubeWhat It Measures
GlucoseGold SST or GrayBlood sugar level
BUN (Blood Urea Nitrogen)Gold SSTKidney function
CreatinineGold SSTKidney function
Sodium (Na+)Gold SSTElectrolyte balance
Potassium (K+)Gold SSTElectrolyte balance
Chloride (Cl-)Gold SSTElectrolyte balance
CO2 (Bicarbonate)Gold SSTAcid-base balance
CalciumGold SSTBone/metabolic function

Comprehensive Metabolic Panel (CMP) — 14 Tests

  • All 8 BMP tests PLUS:
  • Albumin, total protein, alkaline phosphatase (ALP), ALT, AST, and total bilirubin
  • Assesses liver function in addition to kidney function and electrolytes

Complete Blood Count (CBC)

TestTubeWhat It Measures
WBC (White Blood Cells)Lavender (EDTA)Infection/immune response
RBC (Red Blood Cells)Lavender (EDTA)Oxygen-carrying capacity
Hemoglobin (Hgb)Lavender (EDTA)Oxygen transport protein
Hematocrit (Hct)Lavender (EDTA)Percentage of blood that is RBCs
Platelet countLavender (EDTA)Clotting ability
DifferentialLavender (EDTA)Types of WBCs (neutrophils, lymphocytes, etc.)
MCV, MCH, MCHCLavender (EDTA)RBC size and hemoglobin content

Lipid Panel

TestTubeFasting?
Total cholesterolGold SSTRecommended
HDL cholesterolGold SSTRecommended
LDL cholesterol (calculated)Gold SSTRecommended
TriglyceridesGold SSTYes (most affected by fasting)

Coagulation Panel

TestTubeWhat It Measures
PT (Prothrombin Time)Light blue (citrate)Extrinsic coagulation pathway; monitors warfarin
INR (International Normalized Ratio)Light blue (citrate)Standardized PT ratio
PTT / aPTT (Activated Partial Thromboplastin Time)Light blue (citrate)Intrinsic coagulation pathway; monitors heparin
FibrinogenLight blue (citrate)Clotting factor I level
D-dimerLight blue (citrate)Fibrin degradation products (DVT/PE screening)

Quality Assurance (QA)

Quality assurance encompasses all activities that ensure reliable laboratory results throughout the total testing process:

Three Phases of the Testing Process

PhaseActivitiesCommon Errors
Preanalytical (before testing)Patient ID, specimen collection, labeling, transport, processingPatient misidentification, hemolysis, wrong tube, mislabeling, transport errors
Analytical (during testing)Instrument calibration, reagent quality, running controls, test performanceEquipment malfunction, expired reagents, human calculation errors
Postanalytical (after testing)Result reporting, interpretation, documentation, critical value notificationReporting errors, delayed results, wrong patient association

Key Fact: The preanalytical phase accounts for 46-68% of all laboratory errors — this is the phase that phlebotomists directly control.

Delta Checks

A delta check compares a patient's current test results with their previous results:

  • If the current result differs significantly from the historical result, the system flags it for review
  • Delta checks help identify:
    • Specimen mislabeling (results don't match the patient's history)
    • Significant clinical changes (genuine medical deterioration or improvement)
    • Technical errors (wrong tube, hemolysis, contamination)

Lab Requisition Forms

Every blood collection must be accompanied by a lab requisition form (paper or electronic) that includes:

Required InformationDetails
Patient nameFull legal name
Date of birth / MRNPatient identifier
Ordering physicianWho ordered the tests
Tests requestedSpecific tests or panels
Date/time orderedWhen the order was placed
Diagnosis / ICD codeMedical justification for the test
PriorityRoutine, STAT, ASAP, timed
Special instructionsFasting, timed draw, special handling
Specimen sourceVenous, arterial, capillary

Quality Control (QC) vs. Quality Assurance (QA)

Understanding the difference between QC and QA is important for the exam:

ConceptDefinitionExamples
Quality Control (QC)Specific procedures that monitor the accuracy of the testing process itselfRunning control samples, calibrating instruments, checking reagent lot numbers
Quality Assurance (QA)The comprehensive program that monitors the ENTIRE testing process from specimen collection to result reportingPatient ID verification, specimen labeling audits, turnaround time monitoring, proficiency testing, delta checks

Preanalytical Quality Indicators

IndicatorTargetHow to Monitor
Patient identification errors0% (zero tolerance)Wristband audits, labeling audits
Specimen rejection rate<1% of total specimensTrack rejection reasons monthly
Hemolysis rate<2% of specimensMonitor HIL index on analyzers
Specimen labeling errors0% (zero tolerance)Random audits, barcode verification
Turnaround time (TAT)Per test-specific targetsTrack from collection to result
Recollection rate<1%Monitor reasons for recollection
Patient complaintsMinimizeTrack and investigate all complaints

CLIA (Clinical Laboratory Improvement Amendments)

CLIA regulates all clinical laboratory testing in the United States:

CLIA CategoryComplexityExamplesRequirements
Waived testsLow complexityGlucose meters, urine dipsticks, rapid strep, pregnancy testsManufacturer instructions; minimal oversight
Moderate complexityModerateAutomated hematology, chemistry, urinalysisDocumented training, QC, proficiency testing
High complexityHighManual cell differentials, flow cytometry, molecular testingAdvanced training, extensive QC, proficiency testing

Key Fact: Many POCT (point-of-care) tests that phlebotomists may perform are CLIA-waived, but they still require proper training, quality control, and documentation.


Critical Values

Critical values (also called panic values or alert values) are test results that indicate a life-threatening condition requiring immediate physician notification:

Common Critical Values

TestCritical LowCritical High
Glucose<40 mg/dL (hypoglycemia)>500 mg/dL (hyperglycemia)
Potassium<2.5 mEq/L>6.5 mEq/L
Sodium<120 mEq/L>160 mEq/L
Hemoglobin<7 g/dL>20 g/dL
Platelet count<20,000/mcL>1,000,000/mcL
WBC<2,000/mcL>30,000/mcL
PT/INRINR >5.0
TroponinAbove reference range (varies by assay)

Critical Value Reporting Protocol

  1. Verify the result — rerun the test if there is any question
  2. Call the ordering physician or authorized provider within 30 minutes of result verification
  3. Document the following:
    • Date and time of the call
    • Who was notified (name and title)
    • The critical value reported
    • Read-back confirmation (the recipient must repeat the value back)
  4. If the physician cannot be reached, follow the facility's escalation protocol

Medical Terminology for Phlebotomists

Phlebotomists should know common medical terminology related to blood and laboratory testing:

Blood-Related Prefixes and Suffixes

TermMeaningExample
Hem/o, Hemat/oBloodHematology (study of blood)
Ven/o, Phleb/oVeinVenipuncture, phlebotomy
Arteri/oArteryArterial blood gas
-emiaBlood conditionAnemia (low RBCs), bacteremia (bacteria in blood)
-osisAbnormal conditionThrombosis (blood clot)
-lysisDestructionHemolysis (RBC destruction)
-peniaDeficiencyThrombocytopenia (low platelets)
-cytosisIncrease in cellsLeukocytosis (high WBCs)
-stasisStoppingHemostasis (stopping of bleeding)
-centesisPuncture to withdraw fluidAmniocentesis, arthrocentesis

Common Abbreviations

AbbreviationMeaning
CBCComplete blood count
BMPBasic metabolic panel
CMPComprehensive metabolic panel
PTProthrombin time
PTT / aPTT(Activated) partial thromboplastin time
INRInternational normalized ratio
ESRErythrocyte sedimentation rate
GTTGlucose tolerance test
ABGArterial blood gas
H&HHemoglobin and hematocrit
STATImmediately (from Latin "statim")
QNSQuantity not sufficient
POCTPoint-of-care testing
FBSFasting blood sugar
PKUPhenylketonuria

Point-of-Care Testing (POCT)

Point-of-care testing (also called bedside testing or near-patient testing) provides rapid results at or near the patient:

Common POCT Tests

TestMethodTurnaround
GlucoseGlucometer (finger stick)5-10 seconds
Hemoglobin / HematocritHemoCue or i-STAT1-2 minutes
INR / PTCoaguChek1-2 minutes
HbA1cDCA Vantage5-6 minutes
Blood gasesi-STAT or ABL2-3 minutes
Pregnancy (hCG)Urine or blood test strip3-5 minutes
Strep ARapid antigen test5-10 minutes
TroponinPoint-of-care immunoassay10-15 minutes

POCT Quality Requirements

  • Operator training required — documented competency assessment
  • Quality control (QC) must be run per manufacturer guidelines (daily, with each new lot, etc.)
  • Results must be documented in the patient's medical record
  • Proficiency testing — POCT instruments must participate in external quality assessment programs
  • Instrument maintenance — cleaning, calibration per manufacturer schedule
Distribution of Laboratory Errors by Phase (Approximate)
Test Your Knowledge

How long must a gold SST tube clot before centrifugation?

A
B
C
D
Test Your Knowledge

Which phase of the total testing process accounts for the MOST laboratory errors?

A
B
C
D
Test Your Knowledge

What is the MOST COMMON reason for specimen rejection by the laboratory?

A
B
C
D
Test Your KnowledgeMulti-Select

Which of the following tests are included in a Basic Metabolic Panel (BMP)? (Select all that apply)

Select all that apply

Glucose
Complete blood count (CBC)
Sodium (Na+)
ALT (liver enzyme)
Potassium (K+)
Creatinine
Test Your Knowledge

A delta check flags a patient whose potassium level is 7.2 mEq/L today but was 4.1 mEq/L two days ago. This could indicate:

A
B
C
D
Test Your KnowledgeMatching

Match each test panel to the tube color required for collection:

Match each item on the left with the correct item on the right

1
Complete Blood Count (CBC)
2
Basic Metabolic Panel (BMP)
3
PT / INR (Coagulation)
4
Fasting Glucose
Test Your KnowledgeOrdering

Arrange the steps for processing a gold SST tube in the correct order:

Arrange the items in the correct order

1
Centrifuge the tube at the appropriate speed
2
Allow the tube to clot for 30 minutes at room temperature
3
Aliquot serum into a secondary tube if needed
4
Collect the blood and gently invert 5 times
5
Verify the gel barrier has formed between serum and cells
Test Your KnowledgeFill in the Blank

The preanalytical phase accounts for approximately ___% of all laboratory errors.

Type your answer below

Test Your Knowledge

What is aliquoting?

A
B
C
D
Test Your Knowledge

What is the main difference between serum and plasma?

A
B
C
D
Test Your Knowledge

A laboratory reports that a specimen has a high "H" index. This indicates:

A
B
C
D
Test Your KnowledgeMulti-Select

Which of the following are reasons a laboratory would REJECT a specimen? (Select all that apply)

Select all that apply

Hemolyzed specimen
Specimen collected in the correct tube
Clots found in a lavender (EDTA) tube
Specimen labeled at the bedside
Unlabeled specimen tube
Under-filled light blue (citrate) tube
Test Your Knowledge

A physician calls the laboratory because a patient's potassium level is reported as 7.0 mEq/L. This value is:

A
B
C
D
Test Your Knowledge

Which test is MOST affected by the patient not fasting, potentially causing lipemia?

A
B
C
D
Test Your Knowledge

A plain red-top tube (no additive) must clot for how long before centrifugation?

A
B
C
D
Test Your KnowledgeFill in the Blank

Quality control in the laboratory refers to specific procedures that monitor the _____ of the testing process.

Type your answer below

Test Your KnowledgeOrdering

Arrange the three phases of the total testing process in the correct order:

Arrange the items in the correct order

1
Postanalytical phase (result reporting and interpretation)
2
Analytical phase (running the test on the instrument)
3
Preanalytical phase (collection, labeling, transport, processing)
Test Your Knowledge

Which of the following POCT (point-of-care) tests provides results in under 10 seconds?

A
B
C
D
Test Your Knowledge

Which CLIA category includes most glucose meters and urine dipstick tests?

A
B
C
D
Test Your KnowledgeMatching

Match each laboratory abbreviation to its meaning:

Match each item on the left with the correct item on the right

1
QNS
2
STAT
3
POCT
4
HIL
Test Your Knowledge

What information must be included in a critical value notification to a physician?

A
B
C
D
Test Your Knowledge

A specimen tube arrives at the laboratory with a label that reads "John Smith, DOB 3/5/1960." However, the requisition form shows "John Smith, DOB 3/15/1960." What should the lab do?

A
B
C
D
Test Your KnowledgeMulti-Select

Which of the following tests are included in a Comprehensive Metabolic Panel (CMP) but NOT in a Basic Metabolic Panel (BMP)? (Select all that apply)

Select all that apply

Sodium (Na+)
ALT (liver enzyme)
Total protein
Glucose
Albumin
Total bilirubin
Test Your Knowledge

A clot is found in a lavender (EDTA) tube submitted for a CBC. What is the MOST likely cause?

A
B
C
D
Test Your Knowledge

Which of the following is NOT a standard component of a CBC (Complete Blood Count)?

A
B
C
D
Test Your KnowledgeOrdering

Arrange the following steps for handling a STAT specimen in the correct order:

Arrange the items in the correct order

1
Transport the specimen to the laboratory by hand immediately
2
Label the specimen at the bedside
3
Collect the blood following proper technique
4
Verify patient identity with two identifiers
5
Note "STAT" on the specimen label and requisition
Test Your Knowledge

What does the term "hemolysis" literally mean?

A
B
C
D
Congratulations!

You've completed this section

Continue exploring other exams