A laboratory manual on

standard operating procedures

(SOPs)
By
Maram Ali Ali
Clinical biochemist
 Standard Operating procedures
(SOPs)

 SOPs is a set of written instructions that

describe in detail how to perform a laboratory
process or experiment safely and effectively.

 Laboratories must have written SOPs.

 Follow well written SOPs would
ensure
 Consistency:
 Everyone should perform the test exactly the same way.
 So that the same result can be expected from all staff
 Accuray:
 Follow written procedures, help laboratory staff produce
more accurate results.
 They not relying on memory alone so they will not forget
steps in the process.
 Quality :
 Consistency and accurate results are the primary goals of
the laboratories.
 SOPs types

1. Master SOP

2. Administrative SOP

3. Technical SOP:
 Procedure
 Equipment
 Analysis
SOP for collection of blood sampls by
venepuncture
 Objective:
 This SOP is applicable to the collection of blood samples by
venepuncture for the purpose of medical diagnostic testing.

This SOP intends to standardize the venepuncture procedure

for blood sample collected by all related medical staff:
 An authorized prescribers written order of blood sample is required.
 Aseptic technique must be performed.
 Follow sample rejection condition.
 Not more than 2 attempts venepuncture on any one patient by a single
phlebotomist at any one time.
 Follow instruction of blood sampling,
New additive clinical biochemistry tests
 Amylase
 Lipase
 Creatine Kinase (CK)
 Lactate Dehydrogenase (LDH)
 Total iron binding capacity (TIBC)
 Alkaline phosphatase
 Calcium
 Iron
 Sodium
 Potassium
 Magnesium
 Zinc
 Lipase (spectrum)
 Reagent:
 Ready to use
 Stable at 2-8 C up to the expiry date
 Once opened the reagent is stable for 2 months at 2 – 8 C
 calibrator:
 Reconstituted carefully with distilled water (stated on the vial).
 Close vial carefully and allow to stand for 30 min with occasional
swirling.
 Avoid foaming
 Do not shake
 Divided to aliquots stored at -20C for 30 days
 Lipase (spectrum)
  Sample/Calibrator
Reagent 1 500

 samples: Sample/Calibrator 10

Mix, incubate 5 min at 37

 Serum free hemolysis
Reagent 2 125
 Heparin plasma
Mix, incubate 2 min at 37 read A1, 2 min later
 Satability : 24 hrs at 15 – 25 5 days at 2-8 1 year at -20
read A2
 Procedure:
 Wave length 580
 Assay type Kinetic Fixed rate
 Temperature 37
 Zero adjustment against air
 Sensitivity (detection limit) 3 U/L
 Linearity 300 U/L (serum diluted 1:1 sodium chloride)
 Lipase (spectrum)
 Expected value:
 < 60 U/L

 Interfering substances:
 Bilirubin not interfere up to 60 mg/dl.
 Triglyceride not interfere up to 1000 mg/dl

 Safety :
 Do not ingest or inhalation. In case of contact with eyes or skin, rinse immediately with
plenty of soap and water. In case of severe injuries; seek medical advise immediately.
 Amylase (spectrum)
 Reagent:
 Ready to use
 Stable at 2-8 C up to the expiry date
 Once opened the reagent is stable for 2 months at 2 – 8 C
 Reagent 2 contains potassium thiocynate harmful if swalled
 Do not use alpha amylase reagents in case of particulate material
or if absorbane > 0.6.

 Saliva and skin contain alpha amylase: never pipette by mouth and avoid skin
contact with the reagents (use gloves)
 Amylase(spectrum)
  Sample
Reagent 1 800
 samples: Reagent 2 200

 Serum free hemolysis Mix, incubate 1 min at 37

 Heparin plasma Sample 25
 Urine Mix, incubate 1 min at 37 read absorbance. Read again after
 Satability : 7 days at 2-8 1 month at -20 1, 2, 3 min
 Procedure:
 Wave length 405
 Assay type Kinetic non fixed rate or fixed rate
 Temperature 37
 Zero adjustment against air
 Sensitivity (detection limit) 2 U/L
 Linearity 1500 U/L (serum diluted 1:1 sodium chloride)
 Factor 3060
 Amylase (spectrum)
 Expected value:
 serum or plasma < 100 U/L
 Urine < 450 U/L
 24 hrs Urine < 410 U/ 24 h

 Interfering substances:
 Bilirubin not interfere up to 20 mg/dl.

 Safety :
 Do not ingest or inhalation. In case of contact with eyes or skin, rinse immediately with plenty of soap and
water. In case of severe injuries; seek medical advise immediately.
 Lactate dehydrogenase (LDH) (spectrum)

 Reagent:
 Ready to use
Stable at 2-8 C up to the expiry date
Once opened the reagent is stable for 2 months at 2 – 8 C
Prepare working solution 4volume R1+ 1volume R2
 Working solution is stable for 3 weeks at at 2 – 8 C or 2
days at 15 – 25 C.
Do not use alpha amylase reagents in case of particulate
material or if absorbance > 1.0.
 Lactate dehydrogenase (LDH) (spectrum)

  Sample
Reagent 1 400

Reagent 2 100
 samples:
Sample `10
 Serum free hemolysis
 Heparin plasma Mix, incubate 30 sec at 37 read absorbance. Read again after

 Satability : 4 days at 20 – 25 6 weeks at 2 -8 1, 2, 3 min

 Procedure:
 Wave length 340
 Assay type Kinetic non fixed rate
 Temperature 37
 Zero adjustment against air
 Sensitivity (detection limit) 10 U/L
 Linearity 1200 U/L (serum diluted 1:5 sodium chloride)
 Factor 8059
 Lactate dehydrogenase (LDH) (spectrum)

 Expected value:
 Adult 240 – 480 U/L
 Children:
 Female < 580 U/L
 Male < 1103 U/L
 Pre term < 1103 U/L

 Interfering substances:
 Hemolysis
 Lipemia

 Safety :
 Do not ingest or inhalation. In case of contact with eyes or skin, rinse immediately with plenty
of soap and water. In case of severe injuries; seek medical advise immediately.
 Creatine Kinase (CKMB) (spectrum)
 After MI CK levels begins raising between 4-6 hrs after first acute
symptoms, reaching the peak between 18 – 30 hrs coming back
to normal during third day.

 Reagent:

 Ready to use

 Stable at 2-8 C up to the expiry date

 Once opened the reagent is stable for 1 months at 2 – 8 C

 Creatine Kinase (CKMB) (spectrum)
 samples:
 Serum free hemolysis
 Satability : 2 hrs at 20-25 5 days at 4-8
 Procedure:
 Wave length 340
 Assay type Kinetic non fixed rate
 Temperature 37
 Zero adjustment against air
 Sensitivity (detection limit) 2 U/L
  Sample
 Linearity 2000 U/L (serum diluted 1:2 sodium chloride)
Reagent 1 400
 Factor 1651
Reagent 2 100

Sample `20

Mix, incubate 5 min at 37 then . Read after 1, 2, 3 min

 Creatine Kinase (CKMB) (spectrum)

 Expected value:
 Normal value up to 20 U/L
 Border line 20 - 25 U/L ( follow up is recommended)
 MI >= 25 U/L

 Interfering substances:
 Bilirubin not interfere up to 25 mg/dl.
 Triglyceride not interfere up to 900 mg/dl

 Safety :
 Do not ingest or inhalation. In case of contact with eyes or skin, rinse immediately with
plenty of soap and water. In case of severe injuries; seek medical advise immediately.
 Total iron binding capacity(TIBC) (spectrum)

 Reagent:
 Ready to use
 Stable at 2-8 C up to the expiry date
 Once opened e reagent is stable for 1 months at 2 – 8 C

 calibrator:
 Reconstituted carefully with distilled water (stated on the vial).
 Close vial carefully and allow to stand for 30 min with occasional swirling.
 Avoid foaming
 Do not shake
 Divided to aliquots stored at -20C for 30 days
 Total iron binding capacity(TIBC) (spectrum)
  Calibrator calibrator Sample Sample
blank blank
R1 500 500 500 500

calibrator 40 40    

sample     40 40

Mix and incubate for 5 min at 37

samples:
 Serum free hemolysis R2   150   150
 Morning sample is preferable to avoid lowMix
results.
and incubate for 7 min then read against sample blank
 Satability : 7 days at 20-25 3 days at 2-8 one year at -20

Procedure:
 Wave length 630
 Assay type end point
 Temperature 37
 Zero adjustment against air
 Sensitivity (detection limit) 70 U/L
 Linearity 700U/L (serum diluted 1:1 sodium chloride)
 Total iron binding capacity(TIBC) (spectrum)

 Expected value:
 1 day 134 – 318 ug/dl
 1 weak 190 – 324 ug/dl
 3 – 12 months 290 – 436 ug/dl
 1 – 10 years 262 – 497 ug/dl
 11 – 16 years 290 – 441 ug/dl
 Adult
 female 274 - 497 ug/dl
male 291 - 430 ug/dl
 Interfering substances:
 Bilirubin not interfere up to 30 mg/dl.
 Lipemic samples not recommended
 Albumin > 7 g/dl decrease TIBC

 Safety :
 Do not ingest or inhalation. In case of contact with eyes or skin, rinse immediately with plenty of soap and
water. In case of severe injuries; seek medical advise immediately.
 Alkaline phosphatase (ALP) (Elitech)

 Reagent:
Ready to use

Stable at 2-8 C up to the expiry date

Once opened e reagent is stable for 1 months at 2 – 8 C

 Alkaline phosphatase (ALP) (Elitech)
 samples:
 Serum free hemolysis
 Separate blood sample from erythrocytes as possible, better to analyze freshly specimen (no later than 4 hrs
at room temperature
 ALP activity may increase when refrigerated (2-8) or frozen, if sammple frozen for extended storage move
them to room temperature 18 – 24 hrs before analysis
 Satability : 7 days at 20-25 2 months at -20

  Sample
Reagent 1 400
 Procedure:
Reagent 2 100
 Wave length 405
 Assay type kinetic Sample `10
 Temperature 37 Mix, after 1 min incubate 37. Read absorbance at 1 min
 Zero adjustment distilled water
intervals during 3 min
 Sensitivity (detection limit) 6 U/L
 Linearity 900 U/L (serum diluted 1:1 sodium chloride)
 Factor 2750
 Total iron binding capacity(TIBC) (spectrum)

 Expected value:
 Female < 240 U/L
 Male < 270 U/L

 Interfering substances:
 Bilirubin not interfere up to 36 mg/dl.
 Triglycride not interfer up to 600 mg/dl

 Safety :
 Do not ingest or inhalation. In case of contact with eyes or skin, rinse immediately
with plenty of soap and water. In case of severe injuries; seek medical advise
immediately.
Thanks