UNIVERSITY OF SANTO TOMAS | MEDICAL TECHNOLOGY

CLINICAL CHEMISTRY & MOLECULAR DIAGNOSTICS | SPECIMEN MANAGEMENT

PATIENT IDENTIFICATION

 Use two to three identifiers

 Name, medical record number, date of birth
 Special Situations such as in paternity testing and medicolegal procedures – chain of custody for
the specimen
 Active process
 Patient stating his/her name
 Phlebotomist will verify information via wrist band of patient or through the test requisition
form or computer order
 In case of pediatric patients – parent/guardian should be present and be asked to actively
provide the information
 Submitted samples must be labelled properly by the patinet himself.

BLOOD SPECIMEN

 Types of Blood Sample

 Venous Blood – from vein; usually the specimen of choice
 70 mg/L (0.39 mmol/L) less glucose than capillary blood [about 1.4%)
 0.9% less potassium than capillary blood
 PCO2 is 6-7 mmHg (0.9 kPa) greater than capillary blood.
 Higher levels of bilirubin (5%), Calcium (4.6%), Chloride (1.8%), Sodium (2.3%), and Total
Protein (3.3%) compared to capillary blood content.
 Purplish red
 Arterial Blood
 Bright red
 Capillary Blood
 Mixture of arterial and venous blood
 Plasma
 From anticoagualted blood
 Higher LDH and Total Protein compared to serum values.
 Lower glucose (5.1%), Phosphorous (7.0%), and potassium (8.4%) compared to serum
values.
 Bilirubin, cholesterol, and creatinine values have no significant difference whether from
serum or plasma.
 If used in electrophoresis of proteins, extra band in B region will appear owing to
fibrinogen.
 Serum
 From clotted blood
 Blood Colelction Tubes, Additives, and Preservatives
 Heparin (Green Tube)
 Most widely used anticoagulant for chemistry testing
 Prevents coagulation by accelarating the action of antithrombin II which neutralizes
thrombin
 Naturally occurring anticoagulant
 Sodium Heparin – used in toxicology, lead and trace elements analysis
 Lithium Heparin – for electrolytes; incompatible with folate and lithium assay
 Ammonium Heparin
 Ethylenediaminetetraacetic acid (Lavender Tube)
 Commonly used in Hematology testing
 Chelates divalent atoms such as Calcium and Magnesium
 Used in measurement of intracellualr drugs (immunosuppresants), HbA1c analysis,
isolation of genomic DNA, and in qualitative and quantitative virus determination by
molecular techniques
 Inhibits Creatine Kinase and Alkaline Phosphatase
 Lavender-top tube EDTA
 K3EDTA – liquid; will dilute sample by 1-2%
 K2EDTA – spray dried on tube walls
 Pink-top tube EDTA
 Spray dried K2EDTA
UNIVERSITY OF SANTO TOMAS | MEDICAL TECHNOLOGY

CLINICAL CHEMISTRY | SPECIMEN COLLECTION, TRANSPORT, STORAGE, AND PROCESSING

 Used in immunohematology for ABO grouping, Rh typing, and antibody

screening
 White-top tube EDTA
 EDTA and gel
 Often used for molecular diagnostic testing of plasma
 Sodium Fluoride with Potassium Oxalate (Gray Tube)
 NaF is a weak anticoagulant and is mainly a preservative for glucose and lactate. The
addition of Potassium Oxalate compensates for the anticogulant activity.
 Lactate and Glucose analysis
 Effect is not immediate, glycolysis continues for the first one hour after collection.
 Normally, glucose declines in serum samples at a rate of 100 mg/L (0.56
mmol/L) per hour at 25 degrees Celsius.
 Citrate
 For coagualtion studies – Light Blue Tube (9 parts blood : 1 part citrate)
 Used in hematology when platelet sattelism is suspected
 0.105 M (3.2%) or 0.129 (3.8%) Sodium Citrate
 Proper ratio must be strictly observed because anticogualant activity will be reversed by
the calcium reagent in coagualtion assays and cause falsely elevated clotting times.
 Similar effect produced by polycythemia = falsely elevated results due to small
plasma content.
 Inhibits aminotransferases and ALP
 ACP will be stimulated when the substrate used is phenylphosphate
 For westergren ESR – Black Tube ; 4 parts blood : 1 part buffered sodium citrate

Venipuncture

 Equipments
 Antiseptic Cleaning Solution
 70% Alcohol
 Benzalkonium chloride solution
 2% Iodine / Iodophor
 10% Povidone-Iodine
 Bandages and Gauze Pads
 Collection Tubes
 Gloves
 Markers
 Needle Disposal Containers
 Needle Holders
 Needles
 Most commonly used sizes for adults are 19-22 gauge.
 Usually gauge 20 for adults with normal-sized veins
 In pediatric patients gauge 23-25 needles are used.
 Gauge 23 preferred for pediatric patients.
 The larger the gauge number, the smaller the bore.
 1.5 inches long (3.7 cm)
 Syringes with transfer device
 Tourniquets
 Winged infusion sets
 1 inch needle (2.5 cm) length
 Locating Veins
 Apply the tourniquet 3 to 4 inches above the intended puncture site. (Venous Occlusion)
 Some apply the torniquet 4-6 inches (10-15 cm) above the puncture site.
 When a blood cuff is used it is usually inflated to approximately 60 mmHg (8.0 kPa).
 If dorsal vein is being accessed, no tourniquet is used.
 Do not apply tourniquet over an open sore.
 The torniquet should not be left on longer than 1 minute (slight changes in blood composition).
Marked changes have been observed if tourniquet is applied for 3 mins.
 Hemoconcentration – increase in ratio of formed elements to plasma caused by leaving
the tourniquet on too long or too tight which may alter some test results.
 Hemolysis can occur if the tourniquet is too tight or is left on too long
UNIVERSITY OF SANTO TOMAS | MEDICAL TECHNOLOGY

CLINICAL CHEMISTRY | SPECIMEN COLLECTION, TRANSPORT, STORAGE, AND PROCESSING

 Petechiae – small red spots on the skin as a result of bleeding

 Sites
 Antecubital Fossa Area
 Median Cubital Vein – preferred site
 Cephalic Vein – same side of the arm as the thumb
 Basilic Vein – same side of the arm as the pinky finger
 Wrist and Hand Veins
 Use of Winged infusion set is recommended but the draw is slower and there is
increased risk of hemolysis.
 Palpate Veins using index finger.
 Disinfecting the site
 Use 70% alcohol. Clean site in a circular motion going outwards.
 If drawig for blood culutre clean area with 70% alcohol followed by 2% iodine or an iodophor /
10% povidone-iodine.
 Alcohol must be air-dried before applying iodine or iodophor (about 30 seconds)
 Iodine or iodophor contact time must be 60 seconds / 1 minute to be effective
 Timing
 Important for blood constituents that undergo marked diurnal variation, those that require
fasting, and those used to monitor drug therapy.
 Important as well in relation to specimens for alcohol or drug measurements in association with
medicolegal considerations.
 Blood Collection
 Puncture the site with the needle (beveled up) angled at 15 degrees.
 Proper Order of Draw must be observed if collecting multiple samples.
 Yellow for Blood Culture – 8 inversions
 Royal Blue without additive
 Clear without additive; discard if no royal blue is used
 Light blue with Sodium citrate for coagualtion – 3-4 inversions
 Gold/Red Serum Sepator Tube – 5 inversions
 Red Serum Tube with/without clot activator – 5 inversions
 Green Heparin Tube – 8 inversions
 Tan (glass) with Sodium heparin – 8 inversions
 Royal Blue with Sodium Heparin/Sodium EDTA – 8 inversions
 Lavender/Pearl/Pink with EDTA for CBC or Moelcular Studies – 8 inversions
 Gray with Sodium Fluoride/Potassium Oxalate – 8 inversions
 Yellow Glass with ACD for Molecualr Studies and Vell culture – 8 inversions
 Usual Order of Draw
Yellow for blood culture  Red (glass)  Light Blue  Red Plastic / Yellow SST  Green 
Lavender  Gray  Black for ESR  ACD
 Post Venipuncture Care
 Apply pressure on the site of puncture for 1 minute. If patient is taking blood thinner apply
pressure for 10 minutes.
 Notes
 Composition of first drawn dblood is most representative of the composition of circulating blood
and the least affected by fluid shifts where protein-bound components and other large
molecules will be concentrated.
 First-drawn specimen should be used for calcium analysis and other analytes that are both
protein bound and pertinent to critical medical decisions.
 Pumping of fist before venipuncture must be avoided
 Increases Potassium, Phosphate and Lactate
 Lactate elevations would cause pH to decrease causing the Calcium ions to increase.
 Stress increases cortisol, thyroid-stimulating hormone, and growth hormone.
 In children, may also cause increased plasma glucose concentrations due to adrenal
stimulation and increased activity of serum enzymes of skeleatal muscle origin for
chidlren held in physical restraint.

Capillary Blood Collection