LABORATORY ASSAYS

AMY
1. Amyloclastic measures disappearance of starch
(substrate)

2. Saccharogenic measures the appearance of the

reducing sugars product (Classic method)

3. Chromogenic measure the increasing color or

absorbance due to product formation

4. Continuous Monitoring measures the change in

absorbance of NAD (340nm; pH 6.9)

LPS
1. CHERRY-CRANDALL (Titrimetric)
Substrate: Olive oil
Product: Liberated fatty acids

2. Turbidimetric measures the hydrolysis of fates by its rate

of clearing substrate

3. Colorimetric uses peroxidase or glycerol kinase (pH 8.8)

AST
1. KARMEN (Coupled enzymatic)
Indicator enzyme: Malate dehydrogenase
Measures coenzyme: NADH (340nm; pH 7.3-7.8)

ALT
1. WROBLEWSKI-LADUE (Coupled enzymatic)
Indicator enzyme: Lactate dehydrogenase
Measures coenzyme: NADH (340nm; pH 7.3-7.8)

2. REITMAN-FRANKEL (Colorimetric enzymatic)

Uses: alkaline medium
Detects: 2,4-Dinitrophenly hydrazone (BROWN)

ALP
1. BOWERS AND MCCOMB (continuous monitoring)
Substrate: p-nitrophenly phosphate
Product: p-nitrophenol (405 nm; pH 10.2)

2. BESSEY-LOWRY-BROCK

ACP
1. MODIFIED BOWERS AND MCCOMB (continuous
monitoring)
Substrate: p-nitrophenly phosphate
Product: p-nitrophenol (405 nm; pH 5.0)

2. QUANTITATIVE ENDPOINT

Nitrophenylphosphate = Nitrophenol + Phosphate

Thymolphthalein Monophosphate =
Thymolphthalein + Phosphate

3. CONTINUOUS-MONITORING

Alpha-naphthyl phosphate = Alpha-naphthol + Phosphate

Creatine Kinase (CK)
1. TANZER-GILVARG (Forward Reaction)
Uses: Creatine Kinase; Pyruvate Kinase
Indicator enzyme: Lactate Dehydrogenase
Coenzyme: ADP (coupled in the second reaction)
Measures: NADH (340 nm; pH 9.0)

2. OLIVER ROSALKI (Backward Reaction)

Uses: Creatine Kinase; Hexokinase
Indicator enzyme: G6PD
Coenzyme: ATP (coupled in the second reaction)
Measures: NADPH (340 nm; pH 6.8)

Lactate Dehydrogenase
1. WACKER REACTION (Forward)
Substrate: Lactate
Measures: NADH (340nm; pH 8.3 to 8.9)

2. WROBLEWSKI-LADUE (Backward)
Substrate: Pyruvate
Measures: NADH (340nm; pH 7.1 to 7.4)

3. Alpha-Hydroxybutyrate Dehydrogenase Activity

uses alpha-hydroxybutyrate as substrate which has
more affinity to the H subunit thus making it specific for
LD-1.

GGT
1. SZASZ ASSAY
Substrate: Gamma-Glutamyl-p-Nitroanilide
Measures: p-nitroaniline (405-420nm; pH 8.2)

5-nucleotide
Uses the chemical reaction of 5-NT and detects
ribonucleoside production

G6PD
Uses the chemical reaction catalysed by G6PD in the
body and detects NADPH @ 340nm

Cholinesterase
Substrates:
Acylthiocholine
Acetylthiocholine
Proprionylthiocholine
Succinylthiocholine
Reagent: Ellmans Reagent (Dithiobisnitrobenzoic
acid)
Product: 5-mercapto-2-nitrobenzoic acid @ 410nm

ACE
Detects Hippuric Acid @ 228nm