CC2: ALP/LDH Determination

L-Lactate + NAD < - > Pyruvate + NADH + H

LDH Prinicple

6mL of distilled H2O

How to reconstitute the Dry rgt for LDH

14 days @ 2-8 C (Ref Temp)

8 hrs 18-30 C (Room temp)

Rgt. stability

If abs is > 0.6

What abs to discard LDH rgt

decrease

Liver LDH wil (inc/dec) if frozen

• Oxalates

• Oxamates

• EDTA

What inhibitors that interferes with LDH

NADH

product being measured in LDH

340 nm

Because it involves Redox reactions

Wavelength for LDH Dtr.

1. 1 mL of WR

2. Pre warm at 37 C for 3 mins

3. add 25 uL of Serum

4. Incubate at 37 C for 1 min

5. Get Abs at 340 nm against water blank ( 3 reading with 1 min interval)

   1. Get change in Abs/min

Procedure for LDH

Abs/min x 6592

Converion Factor for LDH

Male: 80-285 IU/L

Female: 103-227 IU/L

ref range for LDH, male and female

until 800 IU/L

Linearity of LDH

2

Dilution factor for LDH

exp. date @ 2-8 C

Stability of WR for ALP

R2

Which reagent is Temp sensitive in ALP WR

5 R1 :1 R2

Rgt. prep for ALP (what ratio?)

1. 1 mL of WR

2. Pre warm at 37 C for 5 mins

3. add 25 uL of Serum

4. Incubate at 37 C for 1 min

5. Get Abs at 405 nm against water blank (3 reading with 1 min interval)

   1. Get change in Abs/min

Procedure for ALP Dtr.

P-nitrophenol

What product is being measured in ALP

Principle of ALP

> 500 mg/dL

(For ALP) Hb interferes at

> 20 mg/dL

(For ALP) Bilirubin interferes at

> 1000 mg/dL

(For ALP) TAG interferes at

1000

Linearity for ALP

x 2187

CF for ALP

x2

DF for ALP

35-123 IU/L

Ref range for ALP