APPENDIX
APPENDIX 1: Composition of
polyvitamin and amino acids in hydrosoluble powder
Vitamin A 20.000.000 UI
Vitamin D3 4.000.000 UI
Vitamin E 10.000 mg
Vitamin B1 2.000 mg
Vitamin B2 2.500 mg
Vitamin B6 2.000 mg
Vitamin B12 5 mg
Vitamin C 10.000 mg
Vitamin K3 2.000 mg
Nicotinic acid 10.000 mg
Folic acid 500 mg
Methionine 20.000 mg
Lysine 30.000 mg
Calcium panthotenate 5.000 mg
APPENDIX 2: Reagents for the
determination of total cholesterol
1 Reagent 1 (buffer)
PIPES pH 6.9 90mmol/L
Phenol 26mmol/L
2 Reagent 2(enzymes)
Cholesterol esterase (CHE) 300U/L
Cholesterol oxidase (CHOD) 300U/L
Peroxidase (POD) 1250U/L
4-Aminophenazone (4-AP) 0.4mmol/L
3 Cholesterol CAL 200mg/dl
PREPARATION
Working reagent (WR): The content of the reagent 2 was
dissolved with the corresponding volume of reagent 1 followed by capping and
mixing gently to dissolve contents. Stability of the WR: 4 months at +2 to
+8°C or 40 days at room temperature.
APPENDIX 3: Reagents for the determination of
Triglycerides
· Reagent 1 (buffer)
GOOD pH 7.5 50mmol/L
p-Chlorophenol 2mmol/L
· Reagent 2(enzymes)
Lipoprotein lipase (LPL) 150000U/L
Glycerol kinase (GK) 300U/L
Glycerol-3-phosphate (GPO) 2500U/L
Peroxidase (POD) 440U/L
4-Aminophenazone (4-AP) 0.1mmol/L
ATP 0.1mmol/L
· Triglycerides CAL 200mg/dl
PREPARATION
Working reagent (WR): The content of the reagent 2 was
dissolved with the corresponding volume of reagent 1 followed by capping and
mixing gently to dissolve contents. Stability of the WR: 6 weeks at +2 to
+8°C or 1 week at room temperature.
APPENDIX 4: Reagents for the determination of HDL cholesterol
Reagent 1 (precipitating reagent)
Phosphotungstic acid 14mmol/L
Magnesium chloride 2mmol/L
The reagent is ready to be used
APPENDIX 5: Reagents for the determination of Total protein
Biuret reagent
NaOH 100mmol/l
Potassium Iodide 16mmol/l
Copper sulphate 6mmol/l
Sodium potassium tartrate 16mmol/l
Blank reagent
Sodium potassium tartrate 16mmol/l
NaOH 100mmol/l
Standard
Protein 60g/l (6.0g/dl)
Reagent handling and preparation
Buiret reagent: add 100mls of distilled water
to one vial of Biuret concentration. Stable for 12 month when sealed and stored
at +15-25°C. Protected from light
Blank reagent: dilute the content of blank
reagent bottle with 200mls of distilled water, rinsing the bottle thoroughly.
Stable for 12 months when sealed and stored at +15-25°C. Standard is ready
for use.
APPENDIX 6: Reagents for the determination of creatinine
1 Reagent 1 (picric reagent)
Picric acid 17.5mmol/L
2 Reagent 2(alkaline reagent)
Sodium hydroxide 0.29mol/L
3 Creatinine CAL 2.0mg/dl
PREPARATION
Working reagent (WR): Equal volume of the
reagent 2 was mixed with the corresponding volume of reagent 1 followed by
capping and mixing. Stability of the WR: 10 days at +15 to +25°C.
APPENDIX 7: Reagents for the determination of AST and ALT
respectively
AST ALT
1 Reagent 1(Buffer/substrate)
TRIS pH 7.8 80mmol/L TRIS pH 7.8 100mmol/L
L-Aspartate 200mmol/L L-Alanine 500mmol/L
2 Reagent 2
(Enzymes/coenzymes/á-Ketoglutarate)
NADH 0.18mmol/L NADH 0.18mmol/L
LDH 800U/L LDH 1200U/L
á-Ketoglutarate 12mmol/L á-ketoglutarate
15mmol/L
MDH 600mmol/L
PREPARATION
Working reagent (WR): The content of Reagent 2 was dissolved
with the corresponding volume of Reagent 1 (buffer substrate) followed by
capping and mixing to dissolve contents. Stability: 21 days at +2 to +8°C
or 3 days at room temperature (+15 to +25°C).
| 
