BIOCHEMICAL

CALCULATIONS

Department of Biochemistry,
University of Johannesburg
These calculations were done in
BIC01B1 and should be revised
 Dilutions (p 14 / p 40)

10-fold : 1 part + 9 parts

10 times : 1 part + 9 parts
10X : 1 part + 9 parts
1:10 : 1 part + 10 parts
1 part + __ parts =  parts
1. Prepare a 1 X solution from a 10 X stock with the
following ratios:

a) 10 X
b) 10 times
c) 10-fold
d) 1:10
1 part + __ parts = 10X

1 part + 9 part = 10X

1. Prepare a 1 X solution from a 10 X stock with the
following ratios:

a) 10X
1 part + 9 parts = 10 parts
b) 10 times
c) 10-fold
d) 1:10 1 part + 10 parts = 11 parts
2. Prepare a 2X solution with the ratios a-d)

a) 2X
b) 2 times
c) 2-fold
d) 1: 2 (x: y)
1 part + __ parts = 2X

1 part + 1 part = 2X
a) 2X
b) 2 times 1 part + 1 parts = 2 parts

c) 2-fold
d) 1:2 (x:y) 1 part + 2 parts = 3 parts
3. Prepare a 5X solution with the ratios a-d)

a) 5X
b) 5 times
c) 5-fold
d) 1:5 (x: y)
1 part + __ parts = 5X

1 part + 4 parts = 5X
a) 5X
b) 5 times 1 part + 4 parts = 5 parts
c) 5-fold
d) x:y 1 part + 5 parts = 6 parts
4. Prepare a 33X solution with the ratios a-d)

a) 33X
1 part + 32 parts = 33 parts
b) 33 times
c) 33-fold
d) x:y 1 part + 33 parts = 34 parts
5. Prepare 10 ml of a 1 X solution from a 10 X stock with the
following ratios, a-d)

1 part + 9 parts = 10X

1 ml + 9 ml = 10 ml

1 part + 10 parts = 11 parts

1 ml + 10 ml = 11 ml
6. Prepare 30 ml of a 1 X solution from a 10 X stock
with the following ratios a-d)

1 ml + 9 ml = 10 ml
X3
___ ml + ___ ml = 30 ml

3 ml + 27 ml = 30 ml
 1:10

1ml + 10 ml = 11 ml
X3
3 ml + 30 ml = 33 ml

Work out the exact volume for < 25 ml

30  11 = 2.72 parts
2.7 ml + 27.2 ml = 30 ml
7. Prepare 250 ml of a 1X solution from a 10X stock with the
following ratios a-d)

1 ml + 9 ml = 10 ml
X 25
25 ml + 225 ml = 250 ml
1:10
1ml + 10 ml = 11 ml
X 22.72
22.72 ml + 227.2 ml = 250 ml

0.72 ml = 720l 0.2 ml = 200 µl

The decimal volumes can be measured out with

an automatic pipette
 Automatic pipettes + tips

100 – 1000 l 10 – 100 l

20 – 200 l
8. Prepare a 2X diluted solution from a 10X stock
solution with the ratios a-d)

10X → 2 X
= 10 ÷ 2
So, the factor is 5 X

1-part 10X + 4-part solvent = 2X dilution of the 10X stock

 11X → 2X
= 11 ÷ 2
The factor is 5.5 X

1 parts 10X + 4.5 parts solvent dilution of the 10X stock

 Assume for 5 ml

C1V1 = C2V2
(10X) (  ml) = (2X)(5 ml)
V1 = 1 ml
1 ml + 4 ml = 5 ml
9. Prepare a 5X diluted solution from a 10X stock
solution with the ratios a-d)

10X → 5X
= 10 ÷ 5
So, the factor is 2 X

1 -part 10X + 1 parts solvent = 5X dilution of the 10X stock

 11X → 5X
= 11 ÷ 5
So, the factor is 2.2

1 parts 10X + 1.2 parts solvent = 5X dilution of the 10X stock

10. Prepare a 33X diluted solution from a 10X stock
solution with the ratios a-d)

10 X → 33 X
= 10 ÷ 33
So, the factor is 0.3 X

0.1 parts of 10X + 0.2 parts solvent = 33X dilution of the 10X
stock
 11X → 33X
So, the factor is 0.33X

0.1 parts 10X + 0.23 parts solvent = 33X dilution of the 10 X

stock
11. Describe how you would make a serial dilution of Z to obtain
the ratios a) 10 x b)100X c) 1000X

(p 41 - 42)

Stock a b c

1 part of stock + 9 parts water (10X)

1 part of a) + 9 parts water (10 x 10) = 100X
1 part of b) + 9 parts water (100X 10) = 1000X
12. Describe how you would make a direct dilution of Z to obtain
the ratios a) 10X b) 100X c) 1000X

Stock 10 X: 1 part of stock + 9 parts water

100X: 1 part of stock + 99 parts water

1000X: 1 part of stock + 999 parts water

Percentage (%) p 15 / p 43
% = _____ grams / ml in a total volume of 100 ml

a) v/v volume to volume = ml to ml

b) w/w weight to weight = g to g

c) w/v weight to volume = g to ml

13. How can the following solutions be prepared?
100 ml of 40% (w/v) NaOH?

40 grams NaOH in a final volume of 100 ml

14. 40 ml of 7% (w/v) solution of NaCl?

7 grams NaCl in 100 ml

÷?
___ grams NaCl in 40 ml

 2.8 grams NaCl in 40 ml

15. 200 ml of 95% (v/v) solution of ethanol?

95ml ethanol in 100 ml

X2
190ml ethanol in 200 ml
16. 1 ml of a 10% (w/v) APS solution?

10 grams APS in 100 ml

÷ 100
0.1 grams APS in 1 ml
18. 800 ml of a methanol:ethanol:dH2O solution (60:25:15 v/v/v)

60 parts methanol
25 parts ethanol
15 parts dH2O

Total = 100 parts

 v/v/v = all units are ml

60 ml methanol
25 ml ethanol
15 ml dH2O

Total = 100 ml
X8

Total volume we need is 800 ml

480 ml methanol
200 ml ethanol
120 ml dH2O

Total = 800 ml solution

18. How do you make 1 ml of a 10% solution in the following
conditions?

a) v/v 0.1 ml to 0.9 ml

b) w/w 0.1 g to 1 g of water Why water? 1L = 1 kg

c) w/v 0.1 g to 1 ml
19. If 25 g NaCl is dissolved into a final volume of 500 ml what
is the % (w/v) of the NaCl solution?

If 25 grams are in 500 ml

 grams in 100 ml – why 100 ml????

If 25 grams are in 500 ml

÷5

5 grams in 100 ml = 5%
Dilutions with Stock solutions
(p 39-40)
 Dilutions: C1V1 = C2V2

C1 is always the highest concentration

Looking for V1 when the question reads
“what amount stock solution is needed”
 20. 60 ml of 70% EtOH is needed from 99% stock EtOH, calculation
how much is needed of the stock?

C1V1 = C2V2
(99%) V1 = (70%) (60 ml)
V1 = 42 ml

Take 42 ml of ethanol stock and make it up to a volume of 60 ml

(or take 42 ml ethanol stock + 18 ml dH2O)

21. If the concentration of HCl stock is 12.2 M, how do I
make up 100 ml of a 0.1 M HCl solution?

C1V1 = C2V2
(12.2 M) V1 = (0.1 M) (100ml)
V1 = 0.82 ml
Take 0.82 ml of 12.2 M HCl stock and make it up to a volume
of 100 ml with dH2O
Acid to water

Take 0.82 ml of 12.2 M HCl stock and add it to

99. 18 ml dH2O
22. The concentration of a stock solution is 0.25 M and 200 ml
of a 50 mM solution is needed for your experiment. How
do you make up the solution for the experiment?

C1V1 = C2V2
(0.25 M) V1 = (50 x 10 -3 M) (200 ml)
V1 = 40 ml ….
23. How many µl of 20% SDS is required to bring 1.5 ml of
solution to 0.5%?

C1V1 = C2V2
(20%) V1 = (0.5%) (1.5 ml)
V1 = 0.0375 ml
V1 = 37.5 l …
Looking for C2 when you pipette stock solutions,
you know the volumes that you took, and you
can work out the final volume in a test tube
24. If 8 ml of dH2O is added to 2 ml of 95% EtOH, what is th
e concentration of the diluted ethanol solution?

C1V1 = C2V2
(95%) (2 ml) = C2(10 ml)
C2 = 19%
25. Calculate the concentrations for your x-axis if the
stock was 1 mg/ml
Tube 2: C1V1 = C2V2

(1 mg / ml) (50 l) = C2 (2400 l)

C2 = 0.02 mg / ml

Tube 3: C1V1 = C2V2

(1 mg / ml) (100 l) = C2 (2400 l)

C2 = 0.04 mg / ml
Calculations for mass and
concentration (p 44)
C = n and n = m
V Mr

Molar (M) = moles

Volume (Litres (L))

moles = mass (grams)

MW (grams/moles)
 m = C x V x Mr
grams = Molar x L x grams/mole
ALWAYS CONVERT TO SI UNITS!
26. How is 200 ml of 0.3 M NaCl prepared? (Mr=58.44 g/mole)

m = C x V x Mr

= (0.3 M)(0.2 L) (58.44 g/mole)

= 3.5 grams
Take 3.5 grams of NaCl, dissolve it in 100 ml dH2O and make it
up to 200 ml with dH2O
m = C x V x Mr
= (0.3 M)(200 ml) (58.44 g/mole)
= 3506.4 grams
= 3.5 KILOGRAMS!!
27. How is 50 ml of 20 mM NaOH prepared? (Mr= 40 g/mole)

m = C x V x Mr
= (20 x 10-3 M)(0.05 L) (40 g/mole)
= 0.04 grams
Take 40 mg of NaOH and dissolve in 25 ml dH2O and make it
up to 50 ml with dH2O
28. How many moles of NaCl are present in 50 ml of 0.15 M
solution?

C=n
V

(0.15 M) = n / 0.05 L
= 7.5 x 10 -3 moles
= 7.5 mmoles
29. Express 2.5 M NaCl (Mr = 58.44 g/mole) as a %

mass = C x V x Mr
= (2.5 M) (0.1 L) (58.44 g/mole)
= 14.61 grams

If 14. 61 grams are in 100 ml = 14.61%

30. What is the molar concentration of 10% NaCl solution?

10 grams in 100 ml
Mass = C x V x Mr
10 g = C (0.1 L) (58.44 g/mole)
C = 1.71 M
31. 0.4 mole HCl is dissolved and diluted to 50cm3
Calculate the HCl concentration

50 cm3 = 50 ml

C=n
V
= 0.4 mole / 0.05L
=8M
 Henderson- Hasselbach Equation
[A-]
pH = pka + log
[HA-]

(Pg 45 -47)
32. You wish to make 2 L of 1 M sodium phosphate buffer,
pH 8.0. How do you make up this buffer? pKa= 6.82

Di-sodium hydrogen phosphate

(Na2HPO4∙12H2O), Mr = 358.14 g/mole
Sodium-dihydrogen phosphate
(NaH2PO4∙2H2O), Mr = 156.01 g/mole
2 L of 1 M Sodium phosphate buffer, pH 8.0

[A-]
pH = pka + log
[HA-]
[HP042-]
8.0 = 6.82 + log [H2PO4-]
[HP042-]
1.18 = log
[H2PO4-]
[HP042-]
15.14 =
[H2PO4-]
15.14 parts Na2HPO4 : 1 part NaH2PO4

ratio HPO4-2 : H2PO4- = 15.14 : 1

total parts = 15.14 + 1 = 16.14

 m= C x V x Mr

m(acid) = 1 x 1 M X 2 L x 156.01 g/mole

16.14

= 19.33 g in 2 L

m(base) = 15.14 x 1 M x 2 L x 358.14 g/mole

16.14

= 671.9 grams in 2 L
Dissolve 19.33 g NaH2PO4∙2H2O and 671.9 g Na2HPO4∙12H2O
into 1 L with dH2O and make up to 2 L with dH2O
33. You wish to make 2 L of 1 M sodium phosphate buffer, pH 8.0;
pKa= 6.82. You have stocks of 1 M NaH2PO4 and 1 M Na2HPO4
How much of each stock should be added together to make up
the desired buffer?
2 L of 1 M Sodium phosphate buffer, pH 8.0

[A-]
pH = pka + log
[HA-]
[HP042-]
8.0 = 6.82 + log [H2PO4-]
[HP042-]
1.18 = log
[H2PO4-]
[HP042-]
15.14 =
[H2PO4-]
15.14 parts Na2HPO4 : 1 part NaH2PO4

ratio HPO4-2 : H2PO4- = 15.14 : 1

total parts = 15.14 + 1 = 16.14

 For Na2HPO4 = 15.14 x 1 M x 2 L = 1.876 L
16.14

For NaH2PO4 = 1
x 1 M x 2 L = 0.124 L
16.14

 Take 1.876 L Na2HPO4 + 124 ml NaH2PO4 to make up 2L of 1 M

Sodium phosphate buffer, pH 8.0
EXCEPTION : Tris-HCl buffer (p 46)
Even though it is called a Tris- buffer, there is no base and
conjugated acid. Tris is only one powder.

E.g., 500 ml of a 4x Separating buffer (1.5 M Tris- HCl, pH 8.8)

m = C x V x Mr
= (1.5 M) (0.5 L) (121.14 g/mole)
= 91.05 grams
Take 91.05 grams of Tris
1. dissolve in 250 ml water
2. adjust the pH to 8.8 with HCl
3. and make up the volume to 500 ml with dH2O
NEW calculations
(p 21)
 Dilution factors (p21)
DF = Total volume
Amount of sample taken

If 0.4 ml of sample is diluted in a total volume of 2 ml.

Calculate the dilution factor:

2.0 ml
0.4 ml = 5X

Thus, the original sample has been diluted 5X

Which is the same as 1 part of sample + 4 parts dH2O = 5 parts
(÷5 x 2)
___ ml sample + ___ ml dH2O = 2 ml
0.4 ml sample + 1.6 ml dH2O = 2 ml
34. 1.5 X

35. 25 ml + 75 ml = 100 ml

36. x:y

37. 25:75 = 1:3

Stock Concentrations
(p 47-48)
Information required: density ( or SG) and %
When there is density ( or SG) and % in a question then

m=xVx%
m =  (grams / ml) x 1000 ml x ____ %
100%
37. Concentrated HCl (Mr = 36.5 grams/ mole) is 37.5% HCl by weight and has a
density of 1.19. Calculate the molarity of the concentrated acid

m=xVx%
m = (1.19 grams/ml) x 1000 ml x 37.5 %
100 %
mass = 446.25 grams

m = C x V x Mr
446.25 grams = C x 1 L x 36.5 g/mole
C = 12.2 M
38. Concentrated hydrochloric acid (Mr = 36.5 g mol-1) is 32%.
Calculate the concentration of HCl if the density is 1.15 g/ml
m=xVx%
m = (1.15 grams / ml) x 1000 ml x 32 %
100 %
mass = 368 grams

m = C x V x Mr
368 grams = C x 1 L x 36.5 g/mole
C = 10.08 M
39. What is the concentration of acetic acid (Mr= 60.1 g mole-1,
d= 1.06 g/cm3)

m=xVx%
m = (1.06 grams / ml) x 1000 ml x 100 %
100 %
mass = 1 060 grams

m = C x V x Mr

1 060 grams = c x 1 L x 60.01 g/mole

C = 17.66 M
40. Make up 155 ml of 5M H2SO4 (Mr= 98.09 g mole-1, d= 1.8 g/cm3 ;
98%)
m=xVx%
m = (1.8 grams / ml) x 1000 ml x 98 %
100 %
mass = 1 764 grams

m = C x V x Mr
1 764 grams = C x 1 L x 98.09
C = 17.98 M
 C1V1 = C2V2

(17. 98 M) (V1) = (5 M) (155 ml)

V1 = 43 ml

Acid to water
Take 43 ml of 17.98 M H2SO4 stock and add it to
112 ml dH2O
 Normality (p 37)

N = nM

Normality = number of displaceable protons x concentration

41. Describe the preparation of 350 ml of 0.5 N HCl

m=xVx%
m = (1.19 grams / ml) x 1000 ml x 37.5 %
100 %
mass = 446.25 grams

m = C x V x Mr
446.25 grams = C x 1 L x 36.5 g/mole
C = 12.2 M
N = nM
0.5 N = (1) M
N = 0.5 M

C1V1 = C2 V2
(12.2 M) V1 = (0.5 M)(350 ml)
V1 = 14.32 ml

Acid to water!
42. What is the normality of 0.3 M KOH?

K+ + OH-

N = nM
N = (1) (0.3 M)
N = 0.3 N
43. What is the concentration of a 0.1 N H3PO4
(= 0.033 M)
44. What is the molarity of 1N Na2CO3 solution?
(= 0.5 M)
45. What is the normality of 0.5 M H2SO4?
(=1 N)
  = ½ ∑ MiZi2
Ionic strength =  = I = r/2
(p 48-49)
46. What is the ionic strength of 0.02 M solution Fe2(SO4)3?

0.02M Fe2 (S04)3 → 2 Fe+3 + 3 S04 -2

I = ½ mizi2

= ½ [2(0.02 M)(+3)2 + 3(0.02 M)(-2)2]

= ½ [2(0.02 M)(9) + (3)(0.02 M)(4)]

= ½ [0.36 M + 0.24 M]
= ½ [0.6 M] = 0.3 M
47. What is the ionic strength of 0.04 M phosphate buffer?

Acid: 0.04 M NaH2PO4 → Na+1 + H2PO4-1

Base: 0.04 M Na2HPO4 → 2 Na+1 + HPO42-

0.04 M Na+1 + H2PO4-1 +2 Na+1 + HPO42-

 0.04 M Na+1 + H2PO4-1 +2 Na+1 + HPO42-

I = ½ mizi2
I = ½ [(0.04 M)(+1)2 + (0.04 M)(-1)2 + 2 (0.04 M)(+1)2 + (0.04 M) (-2)2]

I = ½ [(0.04 M) + (0.04 M)+ (0.08 M)+ (0.16 M)]

I = ½ [(0.32 M)]
I = 0.16 M
48. What is the ionic strength of 0.04 M phosphate buffer
diluted with 0.3 M NaCl?

Acid: 0.04 M NaH2PO4 → Na+1 + H2PO4-1

Base: 0.04 M Na2HPO4 → 2 Na+1 + HPO42-

+ additional salt 0.3 M NaCl → Na+ + Cl-

 Na+1 + H2PO4-1 +2 Na+1 + HPO42- + Na+1 + Cl-1

I = ½ mizi2
I = ½ [(0.04 M)(+1)2 + (0.04 M)(-1)2 + 2(0.0 4M) (+1)2 +
(0.04 M) (-2)2 + (0.3 M) (+1)2 + (0.3 M) (-1)2
I = ½ [(0.04 M) + (0.04 M)+ (0.08 M)+ (0.16 M) + (0.3 M) + (0.3 M)]

I = ½ [(0.92 M)]

I = 0.46 M