problem # 6
{ Manufacturer of Insulin
Problem submitted by:
Professor David Meredith, P. E., Pennsylvania State University–Fayette,
Uniontown, PA
Problem Statement
Bioengineers are responsible for developing processes
such as the recombinant DNA process described here.
They also develop artificial joints and human organs and
medical tools to support surgeons. Chemical engineers
are involved with sizing the process equipment and
developing the flow path through the process. Electrical
and mechanical engineers build the support systems
such as the instrumentation and distilled water supply.
To meet this year’s increase in global demand, your
team is to design a manufacturing facility capable of
producing 1,800 kg/yr of human insulin. You will be
using a recombinant DNA process by growing E. coli
bacteria that contain Trp-LE’-Met-proinsulin. You will
need to provide the proper growth media, air, heat
and agitation. After a batch has matured, you must
homogenize the material then separate the product
using centrifuges, diafilters and chromatography in a
series of steps with specific conditions as you extract and
purify the product. Finally, the insulin crystals must be
freeze-dried into crystals for final distribution.
Figure 6-1 Production Process
Background
Human insulin is a polypeptide containing 51 amino
acids arranged in two chains. The A chain contains 21
amino acids and the B chain includes 30 amino acids.
The two chains are linked by two disulfide bonds.
Human insulin has a molecular weight of 5,734 and an
isoelectric point of 5.4.
In the proinsulin process (see Figure 6-1), the plasmid
circle of DNA (1) that exists in bacteria are genetically
modified to include part of the human DNA (2). This
recombinant plasmid (3) is inserted into Eschericia coli
(E. coli) (4). A large colony of the inoculated E. coli
bacteria cells are fermented to overproduce Trp-LE’-
Met-proinsulin in the form of inclusion bodies (5), which
are recovered and soluablized. Proinsulin is released
by cleaving the methionine linker using CNBr (6). The
proinsulin chain is subjected to a folding process to allow
intermolecular disulfide bonds to form (7), and the C
peptide is then cleaved with enzymes (8) to yield human
insulin (9). (Note – the process described here has been
simplified and several key steps have been omitted.
Don’t try this at home.)
The first step of the process is fermentation ( See
Figure 6-2 ). The nutrients are premixed and added to
deionized water in a large tank with an agitator. When
conditions are acceptable, a seed colony of inoculated
bacteria are added to the tank. The tank temperature
is maintained at 37°C by a water jacket that can be
either heated (early in the cycle when heat loss from
the reactor is greater than the heat produced by the
bacteria) or cooled (later in the cycle when the bacteria
produce more heat than the reactor loses).
Assumptions and Givens
• The manufacturing facility operates around the clock
for 330 days per year. A new batch is initiated every
48 hours resulting in 160 batches per year. This
implies a required cycle production of about 11.3 kg
of insulin crystals per batch. See Figure 6-2.
• A cubic meter is 1,000 liters and a cubic centimeter
(cm3) is equal to one ml.
• A micron is 10-6 meter.
• Each batch requires glucose (C6H12O6) to feed
the bacteria (CH1.8O0.5N0.2) in 30 m3 of broth
to grow to a final concentration of 37 g of dry cell
Nutrient
Premix
Tank
Air
Compressor Filter
Figure 6-2 Fermentation Process
weight per liter of broth. The basic unbalanced
chemical equation is:
C6H12O6 + H2O + NH3 + (seed bacteria)
=> CH1.8O0.5N0.2 (6-1)
• Atomic masses for selected elements are given
below:
Hydrogen=1.008 Carbon=12.01
Nitrogen=14.007 Oxygen=15.999
Sulfur=32.064 Zinc=65.37
Bromine=79.91
51. The mass of glucose (kg) required to grow
one batch of the e. coli bacteria to the final
concentration is closest to:
a. 1,350 d. 18,500
b. 7,500 e. 225,000
c. 4,500
Additional Assumptions and Givens
• In addition to temperature and food (glucose), the
bacteria also need oxygen to grow. The oxygen is
Agitator
Temperature
Control
Jacket
To Cell Recovery
 provided by air, which is 21% oxygen by volume.
The air is compressed by a 50 kW compressor to 7
atmospheres of pressure and is filtered through a
cartridge air filter before being sparged (released as
tiny bubbles) into the bottom of the reactor.
• As the air bubbles rise through the broth, some of
the oxygen dissolves into the broth solution to be
taken up by the bacteria. The bacteria require 0.35
g of O2 per g of living cells each hour.
• The minimum dissolved oxygen concentration to
support growth is 0.2 mg / liter of broth.
• The maximum dissolved oxygen concentration in the
medium is 6.7 mg/liter at 37°C.
• The amount of oxygen required to support the
colony growth is given by:
k (C ∗ −CDO )
X= LA DO
(6-2)
qO 2
where,
L = liter
X = final concentration of cells (dry cell weight) per liter
of broth (g/L ),
kLA = oxygen volume transfer rate
(m3 O2 / m3broth-h)
CDO* = maximum dissolved oxygen (DO) concentration
in the broth (gO2/L)
CDO = minimum DO concentration to
support growth (gO2/L)
qO2 = respiration rate of cells (gO2/gcell –h)
52. The flow rate of air (m3/s) to maintain the broth at
the final cell concentration is closest to:
a. 2.3 d. 81 up heat from the broth and exits the reactor jacket
b. 17 e. 292
c. 70
Additional Assumptions and Givens
• To maintain constant growing conditions throughout
the broth, the aerated mixture is stirred continuously
by a large agitator that looks like a kitchen mixer on
steroids.
• The agitator is 2.5 m in diameter and stirs the tank
at 60 revolutions per minute. The motor input
power, Po (kW), required by this agitator is given by:
Po = 0.7 N3 Da2 V (6-3)
where,
N = speed of the agitator (revolution per second)
Da = diameter of the agitator (m)
V = the volume of the broth in the reactor (m3)
• Because aerated mixtures are less dense than non-
aerated mixtures, the motor power is reduced. The
input, Pg (kW), for the agitator for the bioreactor is
empirically given by:
0.45
§ Po2 NDa3 ·
Pg k ¨
¸ (6-4)
© Q 0.56 ¹
where,
k = proportionality constant (use 0.5)
Q = volumetric flow rate of gas per volume of tank (s-1)
53. When the volumetric flow rate of gas is equal to
15 (s-1), the electrical input (kW) to operate the
agitator is closest to:
a. 71 d. 813
b. 97 e. 2,268
c. 140
Additional Assumptions and Givens
• At the end of the 18-hour fermentation time, the
broth is cooled from 37°C to 10°C to quench the
biological action. This cooling process occurs in
about 30 minutes. Chilled water at 5°C is injected
into the reactor jacket at a flow rate of 50 L/s, picks
to return to the cooling system.
• The density and specific heat of the broth are 1,020
kg/m3 and 3.8 kJ/kg-°C respectively.
• The density and specific heat of the water are 1,000
kg/m3 and 4.19 kJ/kg-°C respectively.
• The energy balance equation is given by:
(ρV cp ∆T )broth = (Qρ ∆t cp ∆T )water
(6-5)
where,
cp = specific heat of the fluid (kJ/kg-°C)
ρ = density of the fluid (kg/m3)
V = volume of the broth (m3)
Q = volumetric flow rate of chilled water (m3/h)
∆t = time duration of chilled water flow (h)
∆T = temperature difference for the fluid before and
after the heat transfer occurs (°C)
54. The average water temperature in (°C) leaving the
reactor jacket during this cooling period is closest to:
a. 6 d. 20 c. 286
b. 8 e. 35
c. 13
Additional Background
Once the broth has been cooled, it is transferred to
a holding tank to allow the fermentation tank to be
cleaned and prepared for the next cycle. To reduce the
volume of material to be processed, the 30,000 liters
of broth are passed through a centrifuge to reduce the
volume by a factor of four. This process also removes
most of the extra-cellular impurities.
Next the thickened cell sludge is diluted with an equal
volume of buffer solution consisting of 94.4% distilled
water, 0.7% EDTA (ethylenediaminetetraacetic acid),
a sequestering agent to prevent further reactions and
2.9% TRIS-Base (trishydroxymethylaminomethane), a
buffering agent. This process facilitates the separation
of the cell debris particles from the inclusion bodies that
contain the proinsulin.
Additional Assumptions and Givens
Figure 6-3a EDTA Structure
From Fermentation
Holding
Tank
Homogenizer
To Solubilization
Centrifuge
Centrifuge
Figure 6-3b. Cell Recovery Process
The structure of EDTA is shown above in Figure (6-3a)
55. The molecular weight of EDTA is closest to:
a. 265 d. 292
b. 276 e. 340
Additional Assumptions and Givens
• A high-pressure homogenizer is used to break
the cells and release the inclusion bodies. Passing
the 15,000 liters of buffered sludge through the
homogenizer three consecutive times through a
circular nozzle at high velocity and under a pressure
change of 80 MPa each time, ruptures the cell walls
to expose the inclusion bodies.
• The homogenizer capacity is 10,102 L/h and the
electrical input to the motor is 225 kW.
• It takes about 1.5 hrs for each pass or 4.5 hrs total
for the process.
• The density of the sludge at this point is 985 kg/m3.
• The pressure change, ∆P (kPa), is given by:
(6-4)
2
∆P = ρV / 2,000
where,
ρ = density of the fluid (kg/m3)
V = throat velocity through the nozzle (m/s)
The flow rate of fluid, Q (m3/s) is given by:
Q = VA (6-5)
where,
V = throat velocity (m/s)
A = cross-sectional area of the nozzle throat (m2)
56. The nozzle diameter (mm) is closest to: η = kinematic viscosity (m2/s) of the fluid
a. 3 d. 17 θ = shape factor (use 1.0 for spherical)
b. 5 e. 22 • The process time, ∆t (s) is given by:
c. 9

Additional Assumptions and Givens Qρ

• The 15,000 liters of homogenized sludge is returned
∆t =
(6-7)

to the centrifuge to separate the inclusion bodies

φ
where,
from the cell debris. The inclusion bodies have a
diameter of 1.0 microns and a density of 1.3 g/cm3.
Q = entering flow volume to be separated (m3)
• Assume water (ρ = 1.0 g/cm3) is the liquid media ρ = density of the liquid media entering the filter (kg/m3)
with a kinematic viscosity of 1.004x10-6 m2/s.
I = throughput (kg/s)
• The centrifuge has an acceleration factor of 1,500g
(one g is equal to an acceleration of 9.81 m/s2), has 57. The length of time (h) required to process each batch
147,718 m2 of effective clarifying surface area and is closest to:
a motor input of 25.6 kW with a yield of 98%. The
centrifuge reduces the volume from 15,000 liters a. 1.8 d. 8.3
to 1,400 liters of slurry at the end of the recovery b. 2.3 e. 46
section. c. 5.3

• The throughput or flow rate of entering fluids to be Additional Background

separated, I (kg/s) is given by: The 1,400 L of inclusion body suspension is transferred to
a glass-lined agitator tank and mixed with small quantities
2 2
d (ρs − ρl )(rω )(V D) Fs of urea and of 2-mercaptoethanol. Urea is a chaotropic
φ= (6-6) agent that dissolves the denatured protein in the inclusion
18ηθ bodies and 2-mercaptoethanol is a reductant that reduces
where, disulfide bonds. It takes a reaction time of 8 hours to
d = particle diameter (m) solubilize 95% of the proinsulin. The next step is to
remove the excess urea and 2-mercaptoethanol with a
ρs = density of the particle (g/cm3) diafilter and replace it with distilled water. This solution
is then filtered through a dead end filter to remove
ρ1 = density of the fluid media (g/cm3) fine particles that might overload filtering processes
downstream. See Figure (6-4).
(rω2) = acceleration factor (m/s2)
V / D = effective clarifying surface area (m2) Additional Assumptions and Givens
• The design flux capacity of the membrane used
Fs = correction factor for fraction of solid present (use is 4.2 liters per hour for each square meter of
0.05 for 20% solids)
membrane.

From Cell Recovery

Distilled Filter To CNBr Cleavage
Water
Agitated Holding
Tank Tank

Reactant
Diafilter

Figure 6-4 Solubilization Process

• The diafiltration process takes 6 hours to complete. and excess reactants are removed by applying a
vacuum and raising the temperature to 35°C (the
• The sizing equation for a diafilter is given by: boiling temperature of CNBr at that pressure).

Q = J A ∆t (6-7) • Assume the absolute pressure is 0.28 atmospheres

at 35°C and you have to boil off 20 kg of CNBr
where, vapor. One atmosphere (atm) is 101 kPa and
assume the Ideal Gas law:

Q = volume of media to be processed (liters)

PV = nRT (6-8)
J = Design flux capacity (l/m2-h) where,
A = membrane area (m2) R = universal gas constant and is 8.314 kPa-m3/[kmol-K]
∆t = process time (h) P = pressure in kPa
58. The membrane area (m2) required for this process is
V = volume in m3
closest to: n = number of kilomoles of gas

a. 0.02 d. 55 T = temperature in degrees Kelvin.
b. 18 e. 590
c. 41 59. The minimum volume (m3) of the CNBr vapor
produced at evaporator conditions is closest to:

Additional Background a. 1.940 d. 54.3

The chimeric protein is cleaved in a well-mixed reactor
with CNBr (cyanogen bromide) in a 70% formic acid
solution into the signal sequence Trp-LE’-Met, which
contains 121 amino acids and the denatured proinsulin
(82 amino acids). The reaction takes 12 hours at 20°C.
The final mass of proinsulin is 31% of the mass of the
initial Trp-LE’-Met-proinsulin. See Figure (6-5).
Additional Assumptions and Givens
• A small amount of toxic cyanide gas is formed as a
byproduct of the cleavage reaction. This toxic gas
b. 9.570 e. 1,800
c. 17.0
Additional Background
A sulfitolysis step is used to unfold the proinsulin, break
any disulfide bonds and add SO3 moieties (molecule
segments) to all sulfur residues on the cysteines. This
reaction occurs in a well-stirred reactor under alkaline
conditions (pH 9 to 11) over a 12-hour period. The
reagents are filtered through a diafilter and replaced
with a 20% by weight solution of HCl –guanidine.

Vacuum
HCI + Guanidine

Evaporator
Holding
Tank
Agitated
Tank

Agitated Reactant
Tank Diafilter

CNBr Cleavage Sulfitolysis

Figure 6-5 Cleavage and Sulfitolysis

 From
Sulfitolysis Filtered
solution
Figure 6-6 Ion Exchange Chromatography
The human proinsulin(S-SO3-)6 is next chromatographically
purified in an ion-exchange column. The bed consists
of small beads coated with a cation (group of atoms
carrying a positive electric charge) exchange resin. An
ionic bond is temporarily formed with the properly
formed proinsulin(S-SO3-)6 molecules as the solution
passes slowly up through the resin. The molecules are
released when the column is regenerated (back-flushed)
with acid. See Figure 6-6.
Additional Assumptions and Givens
• The ion –exchange column measures 140 cm in
diameter with a bed height of 25 cm.
• The 8,000 liters of solution at this point contains 28
kg of proinsulin(S-SO3-)6.
• The maximum holding capacity of the resin is 18 mg
of proinsulin(S-SO3-)6 for each ml of nominal bed
volume. The regeneration cycle begins when the
bed reaches 70% of maximum holding capacity.
Additional Background
Folding of the proinsulin(S-SO3-)6 and disulfide bond
formation takes place in a a well-mixed reactor using
mercaptoethanol to facilitate the disulfide interchange
reaction. The reaction is carried out at 8°C for 12
Distilled
Water
Holding
Agitated
Tank
Tank
Reactant
Refolding Diafilter
Figure 6-7 Refolding and Enzymatic Conversion
Acid Regeneration
To Folding
Proinsulin solution
hours and reaches a yield of 85%. The reactions are
removed and the protein solution is concentrated
with a diafiltration unit followed by purification in a
hydrophobic interaction chromatography (HIC) column
to remove unfolded or incorrectly folded molecules.
The final reaction step is to enzymatically remove the
C peptide from the human proinsulin using trypsin and
carboxypeptidase B. The reaction occurs in a well-mixed
reactor at 10°C for 12 hours. The reagents are removed
by another diafiltration step followed by purification in
another ion-exchange chromatography (IEC) column.
Several additional purification steps are included in
the process. A reversed-phase, high-pressure liquid-
chromotography (PR-HPLC) step removes structurally
similar insulin-like components. A diafiltration process
removes the reactants and concentrates the solution
by a factor of two. The final purification step is a gel
filtration chromatographic column followed by another
diafiltration process to concentrate the solution by
a factor of ten. The 500-liter solution at this point
contains 12.8 kg of insulin. An ultrafilter captures
particles larger than 100 nanometers (>10-7 m). See
Figure 6-7.
The final step is to crystallize the final insulin. The insulin
solution is mixed with ammonium acetate and zinc
chloride in an agitated reaction tank. The crystallization
into insulin6-Zn2 is carried out at 5°C for twelve hours.
IEC RP-HPLC Gel Filter
Distilled
Agitated Holding Water
Tank Tank
Final Purification
Enzymatic Conversion Reactant
Diafilter
The 11.3 kg of insulin crystals are separated in a basket 60. The number of diabetics supported by the insulin
centrifuge and freeze-dried at –20°C under a vacuum produced at this facility is closest to:
(0.0015 atmospheres).
a. 2,100 d. 1.8x106
Additional Assumptions and Givens b. 4,200 e. 4x106
An average diabetic individual’s daily insulin usage is 60 c. 11,500
units. (This quantity varies with body weight, activity
level and diet.)

A unit is 1/22 mg of crystallized insulin.

{ }
Competition
Part I
and
Part II
Solutions
2007
 Problem #6
Manufacture of Insulin

51. Answer a

The molecular weight of glucose is:

(C 6 H 12 O 6 ) = (6x12) + (12x1) + (6x16) = 180

The molecular weight of the bacteria is:

(CH 1.8 O 0.5 N 0.2 ) = (1x12) + (1.8x1) + (0.5x16) + (0.2x14) = 24.6

Six moles of bacteria require one mole of glucose, or

6x24.6 = 147.6g of bacteria require 180g of glucose.

Each batch requires enough glucose to feed the bacteria in 30 m 3 of broth to a

concentration of 37g of dry cell weight per liter of broth.

(37g/lt) (30x10 3 lt) = 1,110 kg of bacteria.

This requires:

1,110kg of bacteria
x 180g of glucose = 1,353 kg of glucose
147.6g of bacteria

52. Answer d

Using equation (6-2), and solving in terms of K LA yields:

(X)(q02 )
K LA =
C*DO − C DO

where,

K LA = Oxygen Volume Transfer (in m 3 02 /m 3 broth - h)

X = final concentration of cells = 37g cells /L

q 02 = Respiration rate of cells = 0.35g 02 /g cell − h

TEAMS 2007
Part I and Part II Solutions 37
 C DO = Minimum DO concentration to support growth = 0.2mg 02 / L

C * DO = Maximum DO concentration in the broth = 6.7mg 02 / L

(37g cell /L)(0.35g02 /g cell − h)(1,000mg02 /g 02 )

K LA =
(6.7mg02 /L − 0.2mg 02 /L)

= 1,992 m 302 /m 3broth − h

Each batch is 30m 3 and the O2 represents 21% of the air:

Also there are 3,600 s/h.

Therefore the Volumetric rate of air required is:

(30m3 /batch)(1,992m 302 /m 3broth − h)

3 3
= 79.04m 3air /s
(3,600s/h)(0.21m 02 /m air )

53. Answer a

Using equation (6-3) with:

N = Speed of the Agitator = 60 rev/min = 1 rev/s

D a = Diameter of the Agitator = 2.5m, and

V = Broth volume in the reactor = 30m 3 , yields:

Ρ 0 = (0.7)(1rev/s) 3 (2.5m) 2 (30m 3 ) = 131.25 kW

Then using equation (6-4) with:

k = 0.5

Ρ 0 = 131.25 kW

N = 1 rev/s

D a = 2.5m, and

Q = 15/s, yields:

TEAMS 2007
Part I and Part II Solutions 38
 0.45
⎡ (131.25 kW) 2 (1rev/s)(2.5m) 3 ⎤
Ρ g = (0.5) ⎢ ⎥
⎣ (15/s) 0.56 ⎦

= 70.2 kW

54. Answer c

ρ broth = Density of broth = 1,020 kg/m 3

C pbroth = specific heat of broth = 3.8 kJ/kg- 0 C

∆T broth = 27 0 C

ρ water = 1,000 kg/m 3

C pwater = 4.19 kJ/kg − 0 C

Q = volumetric flow rate of chilled water (in m 3 /h)

(50L/s)(3,600s/h)
= 50 L/s =
(1,000L/m 3 )

= 180 m 3 /h

30min
∆t = = 0.5h
60min/h

Solving equation (6-5) in terms of ∆ T water yields:

(ρ broth )(30m3 )(Cpbroth )(∆Tbroth )

∆Twater =
(Q)(ρ water )(Cpwater )(∆t)

(1,020kg/m 3 )(30m 3 )(3.8kJ/kg − 0 C)(27 0 C)

=
(180m 3 /h)(1,000kg/m 3 )(4.19kJ/kg − 0 c)(0.5h)

= 8.32 0 C

Water Temperature = 5 0 C + ∆T = 13.32 0 C

TEAMS 2007
Part I and Part II Solutions 39
55. Answer d

The molecular formula for EDTA is:

C 10 H 16 O 8 N 2 and the molecular weight of the molecule is:

(12g x10) + (1g x16) + (8x16g) + (2x14g) = 292 g/mol

56. Answer a

Using equation (6-6) and solving it in terms of V yields:

(∆P)(2,000)
V=
ρ

where,

∆Ρ = Pressure Change = 80 MPa = 80x10 3 kPa

ρ = density of fluid = 985 kg/m 3

therefore,

(80x10 3 kPa)(2,000)
V= = 403.03 m/s
985kg/m 3

Using equation (6-7) and solving it in terms of A yields:

A = Cross-Sectional Area of Nozzle Throat

15,000L/h
Volume 1,000L/m 3
Q= =
Time (90min)(60s/min)

= 2.7x10 −3 m 3 /s

Q 2.7x10 −3 m 3 /s
A= = = 6.892x10 −6 m 2
V 403.03m/s

A = 6.892 mm 2

TEAMS 2007
Part I and Part II Solutions 40
 πD 2
A= , or
4

4A (4)(6.892mm 2 )
D= =
π π

D = 2.96 mm

57. Answer b

d = particle parameter = 1.0 microns = 1x10 −6 m

ρ s = density of the particle = 1.3g/cm 3

ρ 1 = density of fluid media = 1g/cm 3

n = kinematic viscosity of fluid = 1.004x10 −6 m 2 /s

rw 2 = acceleration factor = 1,500g = 1,500 x 9.81m/s 2

V/D = effective clarifying surface area = 147,718m 2

θ = shape factor = 1.0

F s = correction factor for fraction of solid present = 0.05

Using equation (6-8) yields:

(1x10 −6 m) 2 (1.3g/cm 3 − 1.0g/cm 3 )(1,500x9.81m/s 2 )(147,718m 2 )(0.05)

φ=
(18)(1.004x10 −6 m 2 /s)(1.0)

= 1.8 kg/s

Using equation (6-9) yields:

Q = (15,000 lt)(1 m 3 /1,000 lt) = 15m3

ρ= 1.0g/cm 3 = 1.0x10 3 kg/m 3

(15m 3 )(1.0x10 3 kg/m 3 )`

∆t = = 8,333s = 2.3h
1.8kg/s

TEAMS 2007
Part I and Part II Solutions 41
Note:

In reality the density of the entering sludge is 13,600 lt at a density of 1.0g/cm 3 plus
1,400 lt at a density of 1.3g/cm 3 for a total of 15,420 kg in 15,000 lt for a density of
1.03g/cm 3 .

58. Answer d

Solving equation (6-10) in terms of A yields:

Q
A= , where
(J)(∆t)

Q = volume of media to be processed = 1,4000 l

J = Design flux capacity = 4.2 l/m 2 -h

∆ t = process time = 6h

1,400 l
A=
(4.2 l/m 2 − h)(6h)

= 55.55 m 2

59. Answer c

In order to use equation (6-11) we need to calculate (n) the number of kilomoles
of gas. The molecular weight of the CNBr vapor is:

12g + 14g + 80g = 106g/mol = 106 kg/kmol

The removal of 20 kg represents

20kg
n= = 0.189 kmol of gas.
106kg/kmol

Solving equation (6-11) in terms of V yields:

(n)(R)(T)
V=
P

TEAMS 2007
Part I and Part II Solutions 42
 (0.189kmol)(8.314kPa − m3 /kmol−o K)(35 + 273 o K)
=
(0.28atm)(101kPa/atm)

= 17.11 m 3

60. Answer d

Facility production = (1,800kg/yr)(1yr/365 days)(10 6 mg/kg)

= 4,931,506 mg/day

⎛ 1 mg ⎞
Average daily dose in mg = (60 units/day) ⎜ ⎟
⎝ 22 unit ⎠

= 2.73 mg/daily dose

4,931,506mg/day
Number of supported diabetics =
2.73mg/day

= 1,806,412 diabetics

= 1.8x10 6 diabetics

TEAMS 2007
Part I and Part II Solutions 43