SGOT (AST) SGPT (ALT) ALP ACP Lipase Amylase

Method Frankel Reitman Frankel Reitman Bessey, Lowry, Bessey, Lowry, Tietz-Fiereck Caraway, Modified
Brock (PNPP) Brock (PNPP) Turbidimetric
Assay
Blank Distilled Water Distilled Water Distilled Water Distilled Water Distilled Water Distilled Water
(5 minutes before (5 minutes before
reading) reading)
Standard 100 ul of std. 100 ul of std. 50 ul of std. 50 ul of p- 25 ul std. reagent + 250 ul Amylase
reagent + 900 ul reagent + 900 ul reagent + 1500 ul Nitrophenol + 1200 ul lipase substrates + 250 ul
SGOT substrate + SGPT substrate + of alkaline diluent. 1500 ul ACP buffer + 250 ul color developer +
1000 ul color 1000 ul color Mix and read diluent. Mix and Olive oil substrate. 2000 ul de-ionized
(REAGENT BLANK developer (stand developer (stand read Shake vigorously. H2O. Invert to mix
FOR AMYLASE for 20 minutes at for 20 minutes at Stand for 10 and read
DETERMINATION) room room minutes at room
temperature) temperature) temperature
Add 5 ml of 0.4 N Add 5 ml of 0.4 N
NaOH NaOH
Sample 50 ul of serum + 50 ul of serum + 50 ul of serum + 5o ul serum + 250 25 ul serum + 1200 250 ul Amylase
250 ul of SGOT 250 ul of SGPT 250 ul of alkaline ul ACP substrates. ul lipase buffer + substrates + 10 ul
substrate. substrate. substrate. Mix Mix and stand for 250 ul Olive oil of serum + 250 ul
Incubate for 30 Incubate for 15 gently and stand 10 minutes at substrate. Shake color developer +
minutes at 37 C minutes at 37 C for 10 minutes at room temperature vigorously. Stand 2000 ul de-ionized
Add 250 ul of color Add 250 ul of color room Add 1500 ul ACP for 10 minutes at H2O. Invert to mix
developer. Mix developer. Mix temperature. diluent. Mix and room temperature and read
and stand for 5 and stand for 5 Add 1250 ul of read
minutes minutes alkaline diluent.
Add 2000 ul of 0.4 Add 2000 ul of 0.4 Mix and read.
NaOH. Mix and NaOH. Mix and
read read
Wavelength 492 nm 492 nm 405 nm 450 nm 550 nm 600 nm
Standard Value 24 28 100 U/L 15.6 U/L 60 750 U/L

Formula U= (Abs S/ Abs Std) U= (Abs S/ Abs Std) U= (Abs of S/ Abs U= (Abs of S / Abs U = ( Abs of S/ Abs U= ((Abs of rgt. Blk
x 24 U/L x 28 U/L of std) x 100 U/L of std) x 15.6 U/L of Std) x 60 U/L – Abs of S)/ Abs of
rgt. Blk) x 750 U/L)
Normal Value 0-40 U/L 0-30 U/L Infant: 50-165 U/L Male: 4.7-13.6 U/L 30- 200 U/L 60-180 U/L
(sinulat ni ma’am)
41-50 U/L 31-45 U/L Child: 20-150 U/L Female: 5.0-11.0
(Borderline) (Borderline) U/L
Adult= 20-70 U/L 89-134 U/L (nasa
50-400+ (Post 45-100 (Post insert)
Cardiac Infarction) Cardiac Infarction) >60 years old=
30-75 U/L
50-1000+ (Liver 100-2000+ (Liver
Necrosis) Necrosis)
End Color Brown Brown Light yellow Green/light green/ Clear Clear
yellow