Experimental Design and Statistics Workshop Questions

Your completed assignment should only include:

i) calculated p-value value for the statistical test (for hypotheses test questions) and

ii) decision/conclusion on what this means for the statistical significance of the test undertaken.

You do not need to include lists or tables of data directly copied from GraphPad. The above i) and ii) will
be sufficient to demonstrate your ability to use GraphPad Prism correctly.

(1) A number of marathon runners collapse during or at the finish of competitive racing. It is
postulated that these runners are able to run until they collapse because they secrete increased amounts
of endogenous opioids to counteract pain, which would otherwise stop the running well before the
collapse stage. To test this hypothesis, the β-endorphin levels are measured immediately after they
collapse. The -endorphin levels of the same number of runners who did not show signs of extreme
exhaustion at the end of the race were also measured. The results are shown in the Table below. Is th ere
are difference in the secretion of -endorphin between the two groups?
β-endorphin plasma levels (pmol L-1)
Control Runners Collapsed Runners
14 109
20 70
50 47
17 36
45 130
35 108
37 29
16 55
26 42
24 118

(2) In a clinical study, the effects of two new antihypertensive agents (Moriprolol and cardifedipine)
on the lowering of the diastolic blood pressure have been compared. The results are shown in the table
below. Is there a difference between the antihypertensive effects of the two therapeutic agents?

Lowering of Diastolic Blood Pressure (mm Hg)

Subject Moriprolol Cardifedipine
1 12.5 13.5
2 13.0 14.0
3 12.5 12.5
4 17.5 12.0
5 13.5 13.5
6 12.5 14.0
7 17.5 11.5
8 22.5 12.5
9 13.5 13.0
10 12.5 13.0
11 12.0 13.5
12 13.5 14.0
(3) A medical device company wishes to examine the ultimate tensile strength of two catheters.
Using tensile analysis, the ultimate strengths of the two medical devices have been determined and are
shown below.

Catheter A (MPa) Catheter B (MPa)

230 248
245 235
231 257
240 256
255 261
234 244

Using an appropriate statistical test, examine whether there is a difference in the two catheters with
respect to their ultimate tensile strengths.

(4) For registration purposes two formulations of a new semi-solid product containing an anti-viral
agent have been manufactured and placed on stability testing under controlled storage conditions (37°C).
After three months, ten units (2 g) of each formulation were removed and the concentration of drug in
each unit determined using a validated high performance liquid chromatography method. The analytical
results are shown below.

Total concentration of drug (mg) in each unit

Formulation 1 Formulation 2
104.1 102.9
108.2 99.6
108.6 98.1
100.8 104.2
106.5 90.2
101.0 101.0
102.6 99.9
99.2 89.5
95.2 95.5
100.8 98.6

Using an appropriate statistical method, determine whether there is a difference in the mean
concentration of drug in each formulation following the period of storage.
(5) A pharmaceutical company wishes to compare the time taken for the release of 30% of the
original drug loading (t30%) from two pharmaceutical preparations (that contain the same mass of a
therapeutic agent). In this ten tablets of each formulation were selected and the release of drug
monitored using dissolution apparatus. The t30% for the two formulations are shown in the table below:

Preparation One (t30%) (mins) Preparation Two (t30%) (mins)

57 70
84 71
81 79
85 61
75 80
62 61
80 69
76 72
79 88
74 65

Using an appropriate statistical method, conclude whether there is a difference between the drug release
(t30%) from the two preparations. The rationale for the choice of statistical method must be clearly shown.

(6) A post-graduate research student wishes to evaluate the analgesic properties of two analgesic
formulations containing the same concentration of therapeutic agent. Twenty subjects are recruited into
the study. Ten subjects receive a single dose of the first analgesic formulation and the remaining ten
subjects receive a single dose of the second analgesic formulation. After 4 h the subjects are exposed to a
painful (ultrasonic based) stimulus and are asked to record the relative pain of the stimulus using a visual
analogue scale (in which 1 refers to no pain and 10 refers to excruciating pain). The results from the
clinical study are shown below.

Recorded Pain Scores (visual analogue scale) associated with:

Analgesic Formulation 1 Analgesic Formulation 2
5 3
7 5
6 4
4 6
6 5
5 2
9 1
8 6
10 4
7 2

Using an appropriate statistical method, determine whether there is a difference in the analgesic
properties of the two formulations. Your answer must include all assumptions made and also must
include a critical evaluation of the limitations of the experimental design.
(7) A therapeutic agent has been developed for the treatment of migraine, however, it has been
suggested that the efficacy of this agent is greater in elderly patients. Therefore, a clinical study has
designed into which two groups of male volunteers (20 – 30 years old and 60 – 70 years old) are recruited.
Each volunteer received one dose of the therapeutic agent and the pharmacokinetic properties were
determined. The results, expressed as the maximum drug plasma concentration, are shown in the table
below:

Maximum drug plasma concentration (g mL-1)

20 – 30 age group 60 – 70 age group
5.8 2.8
10.0 3.6
3.8 4.5
5.7 6.2
6.8 1.2
4.5 1.8
6.9 3.8
8.9 3.7
12.1 5.9
10.7

Using an appropriate statistical test examine whether there is a difference in the maximum plasma drug
concentration between the two age groups.
(8) The table below shows the drug content (mg) of 20 tablets produced using a modification to an
existing tableting procedure

5.13 5.04 5.09 5.00

4.98 5.03 5.01 4.99
5.2 5.08 4.96 5.18
5.08 5.06 5.02 5.24
4.99 5.17 5.06 5.00

The nominal drug content of the batch of tablets is 5.01 mg

Using an appropriate statistical method, comment on whether (or not) the average drug content of the
batch prepared using the modified method is different from the expected content (5.01 mg)

(9) The data presented below describes the diastolic blood pressure of 15 hypertensive adult males
(age 60 –70 years) prior to, and following the implementation of a novel anti-hypertensive therapy.

Patient Reference Diastolic blood pressure (mm Hg) Diastolic blood pressure (mm Hg) 4
Number prior to implementation of the novel weeks after implementation of the
anti-hypertensive therapy novel anti-hypertensive therapy
1 135 130
2 138 124
3 154 140
4 121 121
5 135 115
6 156 150
7 150 150
8 154 132
9 130 118
10 126 104
11 142 128
12 144 142
13 150 134
14 152 129
15 136 116

Using an appropriate statistical test, conclude whether (or not), the novel anti-hypertensive therapy was
successful
Question 10
Three new oral diabetic-related drug has been produced in three different formulations (coded A, B, C).
They were assessed for their ability to release using the dissolution method outlined in the British
Pharmacopoeia. The release kinetics were recorded in the table below. Is there a difference in the release
kinetics of the three new formulations?

Formulation 1 Formulation 2 Formulation 3

5.82 5.69 5.22
5.39 5.39 4.65
5.33 5.35 5.36
5.96 5.36 5.11
5.78 5.01 4.87
5.88 5.2 4.69
5.99 4.36

Question 11:
A Pharmacy owner wants to compare the number of methadone dispensed from her four stores within
the same city. She is interested in comparing the number of dispensings from the four stores to
determine if they are statistically different. If they are significantly different, she may relocate patients
from stores with lower dispensings to the store with the highest dispensings. This would allow her to
upgrade the facilities at one store. Is there a difference in the number of methadone dispensings between
the four pharmacies?

Monthly methadone dispensings

Store 1 Store 2 Store 3 Store 4

Jan 102 97 89 100
Feb 106 77 91 116
March 105 82 75 87
April 115 80 106 102
May 112 101 94 100
June 106 85 88 100

Question 12
A formulation scientist wishes to examine the effects of compression on the time required for tablet
disintegration. Tablets were manufactured by compression at 10, 15 and 20 tonnes in a pilot scale tablet
press. The times required for disintegration of samples of 6 tablets that have been formulated and
compressed at a defined tonnage were examined according to the method described in the British
Pharmacopoeia and are displayed in the following table. Comment on the effects of compression force on
the disintegration time of the samples of tablets.
Tablet Disintegration time Disintegration time Disintegration time
(mins): 10 tonne (mins): 15 tonne (mins): 20 tonne
compression force compression force compression force
1 15.8 17.7 21.4
2 15.5 18.5 20.8
3 15.9 18.4 20.4
4 16.4 17.7 20.5
5 16.4 18.8 21.8
6 16.4 18.8 21.8
Question 13

You are involved in formulating a controlled-release drug delivery system involving the diffusion of a drug
across a novel porous self-assembling peptide polymeric system, that has the potential to be utilised as a
tablet coating for oral drug delivery. A side by side diffusion apparatus with donor and receptor
compartments, separated by the novel polymer, is utilised. The donor compartment comprises a solution
of the drug at a defined concentration. The receptor contains phosphate buffer solution at pH 7.4 but no
drug. To examine the rate of diffusion of the drug from the donor compartment to the receptor
compartment through the polymeric film (5cm2 surface area), samples of solution from the donor
compartment are removed at defined periods and the mass of the drug is quantified using Reverse Phase
HPLC and UV spectroscopy (see table).
The diffusion of a therapeutic agent across a polymeric membrane is defined by:

Where:
D = the diffusion coefficient of the drug across the polymeric membrane
A = the surface area of the film exposed within the side by side diffusion cell
k = the membrane: donor compartment partition coefficient
h = the thickness of the polymeric film
Cd = concentration of the drug in the donor compartment

= slope

The theory of drug release tells us if the concentration of the drug remains constant (within 90% of the
original concentration) the diffusion of the drug across the membrane will be constant (linear).
The flux (J) of the drug will therefore be defined by:

Where:
S = surface area of the membrane
a) Using an appropriate statistical method calculate the flux of the drug across the polymeric membrane
b) Determine the equation of the line of best fit for mass of drug (mg) relative to time (h)
c) Calculate the Correlation coefficient (r).
d) Calculate the Coefficient of determination (r2)

Time (Hours) Mass of drug in the donor compartment (mg)

1 373.2
2 369.9
3 367.4
4 365.1
5 363.3
6 362.9
7 360.2

Question 14

Data has been collected on Six volunteers during a Phase I clinical trial
The investigator is interested in determining if there is a correlation between the subject’s weights and
the cumulative amount of drug excreted in the urine after 24 hours (see table below)
a) Determine the equation of the line of cumulative amount of drug excreted in the urine after 24 hours
relative to mass (kg)
b) Calculate the Correlation coefficient (r).
c) Calculate the Coefficient of determination (r 2).

Subject Weight (Kg) Cumulative

amount of drug
excreted in the
urine after 24
hours (mg)
1 67.0 732

2 107.7 501

3 98.1 467

4 89.0 487

5 83.0 491

6 74 651
Question 15

You are a scientist working in a clinical trial company. You have supplied with dosage data (C max
measurements) for a new tablet formulation of atenolol (Formulation B). It is hoped that this will provide
the same overall maximum concentration (Cmax) as the original atenolol tablet (Formulation A) but at a
reduced manufacturing cost.
Atenolol Formulation A Atenolol Formulation B

Cmax (ng/ml) Cmax (ng/ml)

59 64

61 55

62 61

70 55

57 54

84 60

77 83

54 61

71 54

64 66

Use a statistical test to compare the Cmax measurements of the atenolol formulations. Is formulation B a
suitable alternative based on these dosage data measurements?