Methodologies Of Chemistry Tests

Front 1

Albumin (BCR)

Back 1

Reagents: bromcresol green, hydroxymethyl aminomethane and succinic acid
A dye binding method in which bromcresol green (anionic dye) binds to albumin.
Reference range: 3.4—5.0g/dL

Front 2

Alkaline Phosphatase

Back 2

Reagent 1: ρ-Nitro phenyl phosphate; R2: Mg ions, AMP & activators; pH @ 10+/-.1
p-NPP + H2O → alk phos → p-Nitro phenol + H3PO4
Reference range Adults: 30 - 90 U/L (Children have higher normal values)

Front 3

Alanine aminotransferase (ALT)

Back 3

Reagents: α-Ketoglutaric acid, L-alanine, NADH, Lactate dehydrogenase (LDH)
glutamate.
Reference range: 6 to 37 U/L @ 37◦C;

Front 4

Amylase

Back 4

A chromogenic substrate, 2-chloro-p-nitrophenyl-α-D-Maltotrioside (10 CNPG3), is hydrolyzed by α-amylase
Reference range: Serum 30-100 U/L
Urine 1-7 U/L

Front 5

Aspartate aminotransferase (AST)

Back 5

Modified Karmen method.
Reagent: α-Ketoglutarate, L-aspartic acid, NADH, Lactate dehydrogenase (LDH), Malate dehydrogenase (MDH)
Reference values: 5 to 30 U/L @ 37◦C

Front 6

BUN (Blood Urea Nitrogen)

Back 6

Urea + H20→Urease→2NH3 + CO2→ NH3 + alpha-ketoglutarate + NADH → Glutamate Dehydrogenase (GD)→L- glutamate + NAD + CO2
Reference range: 7-18mg/dL

Front 7

Calcium

Back 7

Calcium reacts with o-Cresolphthalein Complexone in an alkaline medium to form a purple-colored complex.
Reference range: 8.60 - 10.0 mg/dL

Front 8

Cholesterol

Back 8

Cholesterol Esterase -- releases free cholesterol; Cholesterol Oxidase -- oxidation (H202 formed); 4-Amino-antipyrine, peroxidase, Quinonimine dye = indicator reaction
Reference range < 200mg/DL

Front 9

CK-MB isoenzyme

Back 9

Anti-human CK-M antibody (Mouse), enough to inhibit up to 2000 U/L of CK-MM.
ADP, G6PDH, Creatine phosphate, NAD, D-Glucose and adenylate kinase inhibitors.
Reference values: 0-10 U/L; %CK-MB < 6%

Front 10

Creatinine

Back 10

A modified Jaffe procedure
Creatinine + Picric acid → alkaline soln→ Creatinine-picrate complex
Reference range -- men: 0.9-1.2mg/dL; Women 0.6-1.1mg/dL

Front 11

Direct Bilirubin

Back 11

modified Jendrassik-Grof rxn
Sulfanilic acid + sodium nitrite→diazo
Diazo + bilirubin + dimethyl sulfoxide (DMSO)→azobilirubin (blue-green)
Reference range: 0.2-1.0 mg/dL

Front 12

λ-glutamyl transferase (GGT)

Back 12

The substrate used is a 3-carboxyl derivative (GLUPA-C) upon which the λ-glutamyl transferase (GGT) acts as a catalyst for the transfer of a glutamyl group onto glycylglycine.
Reference: ♂ ~ 10-34 U/L; ♀ 9-22 U/L

Front 13

Clinitest®

Back 13

Reagent: copper sulfate, sodium carbonate, sodium citrate and NaOH.
10 drops of H20 w/ color change @15 secs, Negative = blue/green color, positive = orange/brown. Also test for galactosemia.

Front 14

Glucose

Back 14

D-glucose + H2O+O2 →(glucose oxidase)→H2O2 + D-gluconate. Then, H2O2 + 4-aminoantipyrine (4-AAP) + Phenol→(peroxidase)→Quinoneimine dye + H2O

Front 15

Glucose reference (variable among labs)

Back 15

• Fasting 65 - 105 mg/dL ( ♀ lower than ♂ )
• Random 65 - 126 mg/dL
• Postparandial < 126 - 135 mg/dL

Front 16

HDL-C

Back 16

Dextran sulfate + Mg2+ reacts with lipoproteins (mostly ApoB-100) → precipitates LDL & VLDL; 2nd reagent (enzymatic) Cholesterol Esterase releases free cholesterol; Cholesterol Oxidase - oxidation rxn (H202 formed); yeilds 4-Aminoantipyrine, peroxidase, & Quinonimine dye.
Reference > 60 mg/dL = desirable;
< 35 mg/dL = heightened risk for CAD

Front 17

Calculated LDL-C

Back 17

Application of the Friedewald Formula allows for a calculated LDL-C:
LDL-C = Total Cholesterol – (HDL + Triglyceride/5)
Reference: 100-130 mg/dL (desired)

Front 18

Indirect, Bilirubin (unconjugated) --calculated

Back 18

Indirect bilirubin = Total bilirubin – (minus) direct bilirubin
Consideration of unconjugated bilirubin is important for neonates, particularly where hemolytic disease of the newborn may be present.