CC2_Prelim_Lab: AST/ALT Lab Experiment

• L-aspartate

• a-ketoglutarate

Substrates for AST Test

• L-alanine

• a-ketoglutarate

Substrates frr ALT Test

Aspartate Aminotransferase

Full name of AST

Serum Glutamate Oxaloacetate Transaminase (SGOT)

Other name for AST

Tip: AST and you shall ____

Serum Glutamate Pyruvate Transaminase

Other name for ALT

4 part R1 + 1 part R2

Preparation of Working Reagents for AST/ALT

• 5 days @ 20-25 C

• 2 weeks @ 2-8 C

Give the stabilities of Working Rgts for AST/ALT

• Serum

• Heparinized Plasma

Allowed Samples/Specimen for ALT

• Serum

• Heparinized/EDTA Plasma

Allowed Samples/Specimen for AST

3 days @ RT

1 week at 4 C

Stability and Storage of ALT Samples

• 24 days at RT

• 28 days at 4 C

• 1 yr. at -20 C

Stability and Storage of AST Samples

1. 1mL of ALT/AST Working rgt

2. Add 100 ul of Serum sample

3. Mix and incubate at 37 C for 50 seconds

4. Read and record Abs at 340 nm against Water Blank

5. Continue reading for next 2 mins. with 1 min interval (incubate in between readings)

6. Calculate for the Abs/min

Give procedure for AST/ALT

Activity (U/L) = Abs/min x 1746

Formula for calculating the Enzyme activity of AST/ALT

Serum/Plasma: <40 U/L

reference range for AST/ALT

10-250 U/L

Give the Linearity for AST

15-250 U/L

Give the Linearity for ALT

> 500 mg/dL

Interference of Glucose @

> 40 mg/dL

Interference of Ascorbic acid @

> 2 mg/dL

Interference of Pyruvate

Negative Bias from 16 mg/dL

Interference of Unconjugated Bilirubin (B1)

Negative Bias from 11.5 mg/dL

Interference of conjugated Bilirubin (B2)

> 600 mg/dL

TAG Interference @

> 5 mg/dL

Methyl dopa Interference @

x10

Dilution Factor used when it above or below the Linearity

x1746

Conversion factor for AST/ALT

minimum of 3

how many abs. reading should be done fro enzymatic act

Test principle of AST

Test principle of ALT