Professional Documents
Culture Documents
Procedural Recommendations:
Collection, Sampling & Pitfalls
Critical Care Testing =
Whole Blood Analysis
■ Whole Blood Analysis =
Many ‘Variables’ need to be ‘controlled’:
Variables Include:
Include
■ Prolonged contact of serum/plasma with cells
■ ‘Collection-Induced’ Hemolysis
■ Effects of Evaporation
(inc temperature)
■ Anticoagulants: Type/Amount/Mixing
■ Collection/Preparation/Analysis Issues
Prolonged Contact of
Serum/Plasma with Cells
• Most Affected Critical Care Tests:
(if cells left in contact with serum/plasma):
– Glucose
– Lactate
– K+
– Hb
• Blood Gases + Others Analysis (Lytes/Metabolites):
– Collect via Syringe; Cannot separate typically
– Analysis should be within 10-15 minutes ideally; no more
than 30 minutes maximum
– No refrigeration recommended
– Keep Sample Well-Mixed
Cell Contact with Plasma:
Parameter Problems
• Affected Parameters:
– Glucose: (Glycolysis)
• Cells use up Glucose while in tube; lowering Gluc values over time.
• Estimated 5 % Decrease/Hour ‘on cells’
– Lactate:
• Due to cellular use of Glucose over time, and with limited O2 present: Increase
Lactic Acid seen.
• Falsely overly high Lact can be seen on samples > 30 min old from collection.
– K+:
• Samples kept as WB: May miss any Hemolysis present.
• Should use BG analyzer that can detect hemolysis.
– Hb:
• WB is proper sample for correct Hb/Hct testing.
• However, proper mixing of WB sample is required to give accurate results.
• Samples sitting too long may require excessive time for proper mixing.
‘Collection-Induced’ Hemolysis
• Hemolysis:
– “The breaking of the Red Blood Cell membrane with the
cellular contents being expelled into the plasma”.
– May interfere with correct measurement several POC
Parameters including:
• Potassium, Hb, SO2%, LDH, Billirubin
• WB Samples vs Serum/Plasma:
– Difficult to detect Hemolysis on WB sample
• Need to rely on BG instrumentation flagging or excessive K+ value
without underlying symptoms
Hemolysis Causes & Detection
• Common Causes of Hemolysis Include:
– Moisture in the collection tube/syringe
– Mechanical destruction of cells by device
– Freezing of sample (ie: Icing during transport)
– Excessive tourniquet use/time
– Too vigorous mixing of sample container
• WB Samples vs Serum/Plasma:
– Difficult to detect Hemolysis on WB sample
• Need to rely on BG instrumentation flagging or excessive K+ value
without underlying symptoms
Exposure to Room Air
• Parameters Affected:
– pO2
• Room Air = Higher O2 levels than syringe sample for normal
range patients; Lower O2 levels in Room Air for patients
receiving O2
• Exposure to Room Air can either Increase or Decrease
measured pO2
– pCO2
• Room Air = Lower CO2 levels than syringe sample
• Generally exposure will LOWER measured pCO2 value
(although not as severe as O2-exposure)
– pH:
• Effected by pCO2 changes (will change inverse to CO2)
pO2 Exposure Effects
– PO2/SO2:
• Will increase because PO2 of heparin solution is about 20 kPa (150 mm Hg)
• pO2 Details: + 0.53 kPa (+4.0 mm Hg)
■ Blood Gases:
■ Patient should be in steady state of ventilation
■ If on Ventilator – (if possible): Settings should
be unchanged for 30 min. prior to collection
■ Collection must be anaerobic
■ Indwelling catheters/A lines may be used for sampling
■ General Collection Notes:
■ In-line catheters must be thoroughly flushed before sampling
■ A volume equal to 3 x “dead space” must be withdrawn prior to
collection to guard against contamination
■ Heparin drips should be stopped at least 30 min prior to collection
and the line should also be flushed
Syringe Use/Collection
Capillary Samples:
• Shows characteristics of both arterial and venous samples.
• Subject to room air contamination if not collected properly
•
■ Capillary Collection Information:
■ Prior to collection >>Warm skin at the site to 39 - 42 o C for 3 min (if possible)
■ After making a single deep puncture, remove the first drop of blood and allow the blood to flow rather than squeezing it out
■ Place the tip of the capillary into the blood drop
■ Fill capillary without introducing air bubble
■ Seal the end that was in the blood immediately (cap or wax)
■ Add a metal mixing bar/flea from the other end of the tube and seal this end also
Capillary Tube Prior to Analysis
• Electrolytes/Chemistries:
– Arterial & capillary blood acceptable.
– Venous Blood usually chosen
(not acceptable for pO2/SO2)
– Use of tourniquet must be kept to
< 2 minutes (hemostasis).
– No muscular activity at site of collection.
Typical ‘Expected’ Values
(Main Gas Parameters)
■ Note: Values listed are for reference only in this presentation. Typically institutions will determine their own ‘Normal
Values’ based on their location and population.
pO2 83 – 108 mm Hg 35 – 80 mm Hg 30 – 50 mm Hg
pCO2 35 – 45 mm Hg 35 – 50 mm Hg 38 – 50 mm Hg
SO2% 95-99 % 65 – 85 % 60 – 85 %
Oxygen Transport
Determination
A Brief Overview-
Things to Consider….
“QC”: What Is It?
• QC = Quality Control Material:
– Aqueous, protein-based, or blood-based solutions.
– Usually pre-assayed for specific analyzers
• Mean and Range Limits usually provided
– Best materials to use are provided by analyzer
manufacturer
– Used to confirm proper system performance and accuracy
of results
– Designed to ‘mimic’ blood analysis
– System should NOT be used if QC is out of limits
– Proper system ‘calibration’ does NOT mean proper
accuracy of results….
“QC”:
When Should It Be Run?
• How Often?
Intra-Laboratory Comparison
Each group using the same Control Material ( same lot no.)
submits data on a Monthly basis. Each member of the
group (Peer Group) receives a monthly report showing the
performance of their unit relative to the rest of the group.