VLD 421: Veterinary Clinical Biochemistry and Laboratory Diagnosis-II (0+1)

Estimation of Lactate dehydrogenase

METHOD OF ESTIMATION OF LACTATE DEHYDROGENASE

Sample: Serum, Plasma

Method of Estimation : GOD-PAP Method

Principle

• Kinetic determination of Lactate dehydrogenase according to the following reaction.

Pyruvate +NADH + H^{+       LDH}             L-Lactate + NAD⁺

Reagent composition

• LDH -P  R1
  • Tris  buffer, (pH 7.40)    80 mmol/L
  • Pyruvate                           1.6mmol/L

• LDH -P  R2
  • NADH

Mix 4 volumes od R1 with 1 volume of reagent R2. Discard  the working reagent if the blank absorbance is less than 1.0 at 340 nm. 

Laboratory procedure

Working Reagnet : 1000μL

Sample : 10μL

Mix and incubate for 10 min. at 37 ° C. Measure the change in absorbance per minute during 3 minutes.

Calculation

       LDH activity (U /L) = ( Change in absorbance/ min ) x 16030