Clinical Chemistry Review

SPECIMEN COLLECTION AND

CONSIDERATION
• 3 Phases of Laboratory Testing:

• PRE-ANALYTICAL (Patient ID, Venipuncture)

• ANALYTICAL (Processing of sample, instrumentation,
calibration)

• POST-ANALYTICAL (Releasing of sample)

 VENIPUNCTURE
- act of obtaining blood sample from a vein.

• Patient Identification
-most critical and important step in specimen collection
• Mislabeling
- mortal sin
 MAJOR VEINS FOR VENIPUNCTURE
• ANTICUBITAL FOSSA
- most common site for venipuncture
• MEDIAN
- vein of choice; most stationary; most anchor vein
• CEPHALIC
- second vein of choice; less stationary
• BASILIC
- third vein of choice because it is close to the brachial artery and nerve.
 EQUIPMENTS FOR VENIPUNCTURE
• TOURNIQUET – should be tied for not more than 1 minute.
• Prolonged application:
HEMOCONCENTRATION
- ⇧ proportion of formed elements
HEMATOMA
- Accumulation of blood in surrounding tissue

• Apply the tourniquet- 3-4 inches above the site.

 EQUIPMENTS FOR VENIPUNCTURE
• NEEDLE – 1 or 1.5 inches
routine gauge:
Gauge 21
most common:
Gauge 20
blood donation:
Gauge 16

GAUGE is INVERSELY PROPORTIONAL to the BORE.

( The bigger the gauge the smaller the bore)
 EQUIPMENTS FOR VENIPUNCTURE
• EVACUATED TUBE SYSTEM
PARTS:
o 2-way needle
o Tube holder
o Evacuated tubes
 EQUIPMENTS FOR VENIPUNCTURE
• SYRINGE SYSTEM
- used when collecting blood from patients with fragile
veins
(elderly, children)
• BUTTERFLY SYSTEM
- used in children
 PLASMA vs. SERUM

PLASMA SERUM
- HAS FIBRINOGEN -NO FIBRINOGEN

-liquid portion of ANTICOAGULANT - liquid portion of COAGULATED BLOOD

EVACUATED TUBES

• Yellow
- SPS (Sodium Polyanetholsulfunate) – blood culture (microbio)
- ACD (Acid Citrate Dextrose) – phenotyping, paternity testing (BB)

• Light Blue
- Sodium Citrate
 3.2%
 3.8%
 BLOOD TO AC ratio - 9:1
EVACUATED TUBES

• Red
- Glass ( no additive )
- Plastic ( silica particle ) – clot activator

• Gold
- serum separator tube
- thixotropic gel
EVACUATED TUBES

• Green
- sodium heparin
- lithium heparin

• Light green
- plasma separator gel
- lithium heparin
EVACUATED TUBES

• Lavender
-Glass (liquid) – K2
Dipotassium salt : Versene

-Plastic (spray) – K3
Tripotassium salt: Sequestrene
- EDTA
Ethylene Diamine Tetra Acetic Acid
EVACUATED TUBES

• Gray
- Antiglycolytic: Sodium Fluoride
- Anticoagulant: Potassium Oxalate

• Black
- Sodium Citrate:
Blood to AC ratio - 4:1
EVACUATED TUBES

• Pink
-spray-dried k2 EDTA

• White
- EDTA or ACD

• Royal Blue
-Sodium Heparin
-K2 EDTA
EVACUATED TUBES

• Tan
-Glass – Sodium Heparin
- Plastic – K2 EDTA

• Orange
-Thrombin – most potent clot activator
-Clotting time: 5-10 mins.
 CAPILLARY PUNCTURE
Arterial blood, venous blood and tissue fluid
- Length of lancet :
- 75mm

- Depth of lancet:
- 2mm

- Incision depth:
- <2mm

- Sites:
- Lateral Plantar portion of the heel, palmar surface of the 3rd and 4th finger, earlobe
- Orientation:
- against fingerprints
 CONVERSION FACTOR
ANALYTE FACTOR
ALBUMIN, HEMOGLOBIN, TOTAL PROTEIN 10
AMMONIA 0.587
BILIRUBIN 17.1
CALCIUM 0.25
CREATININE 88.4
GLUCOSE 0.5555
MAGNESIUM 0.5
THYROXINE 12.9
URIC ACID 0.0595
 CONVERSION FACTOR
ANALYTE FACTOR
AST 0.0167
HCO3, CL-, Li, K+,Na2+ 1.0
BUN 0.357
CHOLESTEROL 0.026
CREATININE CLEARANCE 0.0167
IRON 0.179
PHOSPHORUS 0.323
TAG 0.323
 QUALITY ASSURANCE
concerned with the total process

• QUALITY CONTROL
- System of ensuring accuracy and precision
- Process of ensuring that analytical results are correct.
 QUALITY CONTROL
BLANK WATER
STANDARD ( COLORLESS ) Most specific analytical solution
Value will tell the concentration
Only 1 analyte
CONTROL ( YELLOW ) Value will determine accuracy and
precision
Resembles patient sample
Many analytes
 QUALITY CONTROL
• ACCURACY
- closeness of measured value to true value (reference range)

• PRECISION
-closeness of measured values to each other (refers to previous result)

• SENSITIVITY
- ability of test to detect the smallest amount of analyte

• SPECIFICITY
-ability of test to detect substances without interferences.
 TYPES OF ERROR
• RANDOM ERROR
- Problem in precision
- Unpredictable; hard to avoid
- Pipetting error
- Mislabeling of specimen
- Temperature fluctuation
- Improper mixing of sample or reagents
- Voltage malfunction
 TYPES OF ERROR
• SYSTEMATIC ERROR
- Problem in accuracy
- Predictable
- Deterioration of reagent
- Improper made standard solution
- Contaminated solution
- Calibration problem
- Failing instrumentation
LEVEY-JENNINGS CONTROL CHART
detects random and systematic error
 WESGUARD RULES
12s 1 control observation exceeding the mean Warning rule
+_2s
13s 1 control observation exceeding the mean ⇧ sensi to RE
+_3s
22s 2 control observation consecutively ⇧ sensi to SE
exceeding the same mean +2s or -2s
R4a 1 control exceeding the +2s and another Detection of RE
exceeding -2s
41s 4 consecutive control observations exceeding Detection of SE
+1s or -1s
10x 10 consecutive control observations falling on Detection of SE
one side or the other of the mean
 ERRORS IN LEVEY-JENNINGS CHART
TREND
Gradual loss of reliability
Control values either ⇧ or ⇩
for 6 consecutive days
CAUSE: deterioration of
reagents
Systematic error
SHIFT OUTLIER
Abrupt change in test Highly deviating values
proficiency
Control values that distribute
on 1 side of the mean for 6
consecutive days
CAUSE: Improper Calibration Cause: Pipetting error
Systematic error Random error