Tools and Materials

 Tools
1. Test tube 12 pieces
2. Beaker glass 250 mL 2 pieces
3. Graduated cylinder 10 mL 2 pieces
4. Pipettes 8 pieces
5. Centrifuge tube 2 pieces
6. Centrifuge mechine 1 item
7. Spectronic 20 1 item
 Materials
1. Blood 1 mL
2. Oxalic acid 1 mL
3. Nelson A 16,8 mL
4. Nelson B 4,2 mL
5. Ba(OH)2 0,3 N 1 mL
6. ZnSO4.7H2O 5 % 2 mL
7. (NH4)2SO4 2 mL
8. Aquades 100 mL
Lanes Work
1. Deproteination of blood

1 mL of blood

Put into centrifuge tube that contain 1 mL

oxalic acid
Shake and mix it well
Add 1 mL of Ba(OH)2 0,3 N

Add 2 mL ZnSO4.7H2O

Add 2 mL (NH4)2SO4
Left it 15 minutes
Centrifuge it 15 minutes at 3,500 rpm
Decanted it

Residue Filtrate
9. take it 1 mL
10, add Biuret reagent

Colorless
2. Determination the glucose level in blood
1 mL of free protein blood

Put into test tube

Add 3 mL Cu-alkalis reagent
Put in boiling water for 15 minutes
Put in cold water until constantly cool
Add 3 drops Arsenomolybdate reagent
Mix it well
Read the absorbance at λ = 660 nm

Absorbance value
3. Making standard curve
1 mL of glucose solution 0,1 ;
0,3 ; 0,5 ; 0,7 ; 0,9 mg/mL

Put into test tube

Add 3 mL Cu-alkalis reagent
Put in boiling water for 15 minutes
Put in cold water until constantly cool
Add 3 drops Arsenomolybdate reagent
Mix it well
Read the absorbance at λ = 660

Absorbance value

4. Blanko solution

1 mL aquades

Put into test tube

Add 3 mL Cu-alkalis reagent
Put in boiling water for 15 minutes
Put in cold water until constantly cool
Add 3 drops Arsenomolybdate reagent
Mix it well
Read the absorbance at λ = 660

Absorbance value
 Reactions :
First Experiment :

 2 (aq) + Ba(OH)2 (aq) → (aq) + H2O (l)

(Murray, 1977)

 (aq) + ZnSO4.7H2O (aq) → 2

2-
(s) + SO4 (aq) + 7H2O (l)

Second Experiment :

 (aq) + Cu2+ (aq) → (aq) + Cu2O (s)

 Cu2O (s) + [MoAs12O40]3- (aq) → 2Cu2+ (aq) + [MoAs12O40]4- (aq)

Third Experiment :
 (aq) + Cu2+ (aq) → (aq) + Cu2O (s)
 Cu2O (s) + [MoAs12O40]3- (aq) → 2Cu2+ (aq) + [MoAs12O40]4- (aq)

Fourth Experiment :
 CuO (aq) + H2 (g) → Cu+ (s) + H2O (l)
 Cu2O (s) + [MoAs12O40]3- (aq) → 2Cu2+ (aq) + [MoAs12O40]4- (aq)
Attachment
Calculation :
 M1.V1 = M2.V2
2 mg/mL.V1 = 0,9 mg/mL.10 mL
V1 = 4,5 mL
 M1.V1 = M2.V2
0,9 mg/mL.V1 = 0,7 mg/mL.10 mL
V1 = 7,78 mL
 M1.V1 = M2.V2
0,7 mg/mL.V1 = 0,5 mg/mL.10 mL
V1 = 7,14 mL
 M1.V1 = M2.V2
0,5 mg/mL.V1 = 0,3 mg/mL.10 mL
V1 = 6 mL
 M1.V1 = M2.V2
0,3 mg/mL.V1 = 0,1 mg/mL.10 mL
V1 = 3,33 mL
Cu Alkalis making :
Nelson A : Nelson B = 4 : 1
Nelson A = 4/5 x 21 mL = 16,8 mL
Nelson B -= 1/5 x 21 mL = 4,2 mL
Data :
C (mg/mL) A
0,1 0,284
0,3 0,380
0,5 0,469
0,7 0,283
0,9 0,384

Graphic :

Absorbance of sample = 0,9849

0,9849 = 0,0515x + 0,3343
0,65039 = 0,0515x
x = 12,629 mg/mL

Comparation Data :

Concentration (mg/mL) A
 0,1 0,288
0,3 0,340
0,5 0,347
0,7 0,383
0,9 0,466
Graphic :