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
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
| Type | Source | Tube | Contains Fibrinogen? | Clotting Required? |
|---|---|---|---|---|
| Serum | Clotted blood | Red top, Gold SST | No (consumed during clotting) | Yes — must clot before centrifuging |
| Plasma | Anticoagulated blood | Green (heparin), Lavender (EDTA), Light blue (citrate) | Yes (fibrinogen preserved) | No — anticoagulant prevents clotting |
Clotting Times Before Centrifugation
| Tube Type | Clotting Time | Reason |
|---|---|---|
| Gold SST (clot activator + gel) | 30 minutes at room temperature | Clot activator accelerates clotting |
| Red top (no additive) | 60 minutes at room temperature | No clot activator; natural clotting process takes longer |
| Anticoagulated tubes (green, lavender, light blue) | No clotting time — centrifuge immediately if needed | Anticoagulant 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
| Feature | Serum | Plasma |
|---|---|---|
| Clotting | Blood must clot first | Clotting is prevented |
| Fibrinogen | Absent (consumed) | Present |
| Tube types | Red, gold SST | Green, lavender, light blue |
| Processing time | Longer (30-60 min clotting + centrifuge) | Shorter (centrifuge immediately) |
| Gel separator | Gold SST | Green/gray PST (plasma separator tube) |
| Common tests | BMP, CMP, lipid panel, thyroid | PT/INR, PTT, stat electrolytes, CBC plasma |
Specimen Integrity Indicators
Modern laboratory analyzers can detect specimen quality issues automatically:
| Indicator | Abbreviation | Appearance | Cause |
|---|---|---|---|
| Hemolysis | H | Pink to red serum/plasma | RBC destruction |
| Icterus | I | Deep yellow/orange serum/plasma | Elevated bilirubin (jaundice) |
| Lipemia | L | Milky/turbid serum/plasma | Elevated 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 Rejection | Description | Prevention |
|---|---|---|
| Hemolysis | Pink/red serum or plasma (ruptured RBCs) — #1 cause of rejection | Proper technique: correct gauge, don't shake, allow alcohol to dry |
| Lipemia | Milky/turbid serum (high fat content) | Ensure proper fasting; note non-fasting on label |
| Clotted anticoagulated specimen | Clots in EDTA, citrate, or heparin tubes | Mix tubes immediately by inversion; check for proper fill |
| Wrong tube type | Test ordered requires a different tube than what was collected | Verify order of draw and tube selection before drawing |
| Insufficient volume | Not enough blood for the test (especially critical for citrate tubes) | Ensure proper tube fill; redraw if underfilled |
| Expired tubes | Tubes past their expiration date may have degraded vacuum or additives | Check expiration dates before use |
| Mislabeled or unlabeled | Cannot confirm patient identity | Label at bedside; use two identifiers |
| Improper transport | Wrong temperature, excessive delay, broken tube | Follow transport protocols for each specimen type |
| QNS (Quantity Not Sufficient) | Sample volume too low to perform the requested test | Collect 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
| Test | Tube | What It Measures |
|---|---|---|
| Glucose | Gold SST or Gray | Blood sugar level |
| BUN (Blood Urea Nitrogen) | Gold SST | Kidney function |
| Creatinine | Gold SST | Kidney function |
| Sodium (Na+) | Gold SST | Electrolyte balance |
| Potassium (K+) | Gold SST | Electrolyte balance |
| Chloride (Cl-) | Gold SST | Electrolyte balance |
| CO2 (Bicarbonate) | Gold SST | Acid-base balance |
| Calcium | Gold SST | Bone/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)
| Test | Tube | What 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 count | Lavender (EDTA) | Clotting ability |
| Differential | Lavender (EDTA) | Types of WBCs (neutrophils, lymphocytes, etc.) |
| MCV, MCH, MCHC | Lavender (EDTA) | RBC size and hemoglobin content |
Lipid Panel
| Test | Tube | Fasting? |
|---|---|---|
| Total cholesterol | Gold SST | Recommended |
| HDL cholesterol | Gold SST | Recommended |
| LDL cholesterol (calculated) | Gold SST | Recommended |
| Triglycerides | Gold SST | Yes (most affected by fasting) |
Coagulation Panel
| Test | Tube | What 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 |
| Fibrinogen | Light blue (citrate) | Clotting factor I level |
| D-dimer | Light 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
| Phase | Activities | Common Errors |
|---|---|---|
| Preanalytical (before testing) | Patient ID, specimen collection, labeling, transport, processing | Patient misidentification, hemolysis, wrong tube, mislabeling, transport errors |
| Analytical (during testing) | Instrument calibration, reagent quality, running controls, test performance | Equipment malfunction, expired reagents, human calculation errors |
| Postanalytical (after testing) | Result reporting, interpretation, documentation, critical value notification | Reporting 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 Information | Details |
|---|---|
| Patient name | Full legal name |
| Date of birth / MRN | Patient identifier |
| Ordering physician | Who ordered the tests |
| Tests requested | Specific tests or panels |
| Date/time ordered | When the order was placed |
| Diagnosis / ICD code | Medical justification for the test |
| Priority | Routine, STAT, ASAP, timed |
| Special instructions | Fasting, timed draw, special handling |
| Specimen source | Venous, arterial, capillary |
Quality Control (QC) vs. Quality Assurance (QA)
Understanding the difference between QC and QA is important for the exam:
| Concept | Definition | Examples |
|---|---|---|
| Quality Control (QC) | Specific procedures that monitor the accuracy of the testing process itself | Running 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 reporting | Patient ID verification, specimen labeling audits, turnaround time monitoring, proficiency testing, delta checks |
Preanalytical Quality Indicators
| Indicator | Target | How to Monitor |
|---|---|---|
| Patient identification errors | 0% (zero tolerance) | Wristband audits, labeling audits |
| Specimen rejection rate | <1% of total specimens | Track rejection reasons monthly |
| Hemolysis rate | <2% of specimens | Monitor HIL index on analyzers |
| Specimen labeling errors | 0% (zero tolerance) | Random audits, barcode verification |
| Turnaround time (TAT) | Per test-specific targets | Track from collection to result |
| Recollection rate | <1% | Monitor reasons for recollection |
| Patient complaints | Minimize | Track and investigate all complaints |
CLIA (Clinical Laboratory Improvement Amendments)
CLIA regulates all clinical laboratory testing in the United States:
| CLIA Category | Complexity | Examples | Requirements |
|---|---|---|---|
| Waived tests | Low complexity | Glucose meters, urine dipsticks, rapid strep, pregnancy tests | Manufacturer instructions; minimal oversight |
| Moderate complexity | Moderate | Automated hematology, chemistry, urinalysis | Documented training, QC, proficiency testing |
| High complexity | High | Manual cell differentials, flow cytometry, molecular testing | Advanced 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
| Test | Critical Low | Critical 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/INR | — | INR >5.0 |
| Troponin | — | Above reference range (varies by assay) |
Critical Value Reporting Protocol
- Verify the result — rerun the test if there is any question
- Call the ordering physician or authorized provider within 30 minutes of result verification
- 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)
- 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
| Term | Meaning | Example |
|---|---|---|
| Hem/o, Hemat/o | Blood | Hematology (study of blood) |
| Ven/o, Phleb/o | Vein | Venipuncture, phlebotomy |
| Arteri/o | Artery | Arterial blood gas |
| -emia | Blood condition | Anemia (low RBCs), bacteremia (bacteria in blood) |
| -osis | Abnormal condition | Thrombosis (blood clot) |
| -lysis | Destruction | Hemolysis (RBC destruction) |
| -penia | Deficiency | Thrombocytopenia (low platelets) |
| -cytosis | Increase in cells | Leukocytosis (high WBCs) |
| -stasis | Stopping | Hemostasis (stopping of bleeding) |
| -centesis | Puncture to withdraw fluid | Amniocentesis, arthrocentesis |
Common Abbreviations
| Abbreviation | Meaning |
|---|---|
| CBC | Complete blood count |
| BMP | Basic metabolic panel |
| CMP | Comprehensive metabolic panel |
| PT | Prothrombin time |
| PTT / aPTT | (Activated) partial thromboplastin time |
| INR | International normalized ratio |
| ESR | Erythrocyte sedimentation rate |
| GTT | Glucose tolerance test |
| ABG | Arterial blood gas |
| H&H | Hemoglobin and hematocrit |
| STAT | Immediately (from Latin "statim") |
| QNS | Quantity not sufficient |
| POCT | Point-of-care testing |
| FBS | Fasting blood sugar |
| PKU | Phenylketonuria |
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
| Test | Method | Turnaround |
|---|---|---|
| Glucose | Glucometer (finger stick) | 5-10 seconds |
| Hemoglobin / Hematocrit | HemoCue or i-STAT | 1-2 minutes |
| INR / PT | CoaguChek | 1-2 minutes |
| HbA1c | DCA Vantage | 5-6 minutes |
| Blood gases | i-STAT or ABL | 2-3 minutes |
| Pregnancy (hCG) | Urine or blood test strip | 3-5 minutes |
| Strep A | Rapid antigen test | 5-10 minutes |
| Troponin | Point-of-care immunoassay | 10-15 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
How long must a gold SST tube clot before centrifugation?
Which phase of the total testing process accounts for the MOST laboratory errors?
What is the MOST COMMON reason for specimen rejection by the laboratory?
Which of the following tests are included in a Basic Metabolic Panel (BMP)? (Select all that apply)
Select all that apply
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:
Match each test panel to the tube color required for collection:
Match each item on the left with the correct item on the right
Arrange the steps for processing a gold SST tube in the correct order:
Arrange the items in the correct order
The preanalytical phase accounts for approximately ___% of all laboratory errors.
Type your answer below
What is aliquoting?
What is the main difference between serum and plasma?
A laboratory reports that a specimen has a high "H" index. This indicates:
Which of the following are reasons a laboratory would REJECT a specimen? (Select all that apply)
Select all that apply
A physician calls the laboratory because a patient's potassium level is reported as 7.0 mEq/L. This value is:
Which test is MOST affected by the patient not fasting, potentially causing lipemia?
A plain red-top tube (no additive) must clot for how long before centrifugation?
Quality control in the laboratory refers to specific procedures that monitor the _____ of the testing process.
Type your answer below
Arrange the three phases of the total testing process in the correct order:
Arrange the items in the correct order
Which of the following POCT (point-of-care) tests provides results in under 10 seconds?
Which CLIA category includes most glucose meters and urine dipstick tests?
Match each laboratory abbreviation to its meaning:
Match each item on the left with the correct item on the right
What information must be included in a critical value notification to a physician?
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?
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
A clot is found in a lavender (EDTA) tube submitted for a CBC. What is the MOST likely cause?
Which of the following is NOT a standard component of a CBC (Complete Blood Count)?
Arrange the following steps for handling a STAT specimen in the correct order:
Arrange the items in the correct order
What does the term "hemolysis" literally mean?
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