Hyun Jung Lee (1310438) RMS 9

RESPIRATORY AND
MUSCULOSKELETAL SYSTEMS (9)
EVALUATION OF INHALER PERFORMANCE
Equivalence Testing of Pressurised Metered Dose Inhalers and the Effect of a Spacer on the
Respirable Dose

 Name: Hyun Jung Lee

 Group: B
 Student Number: 1310438
 Date of Experiment: 5th March 2015
 Supervisor: Anna Morgan

INTRODUCTION

Having been in use since the 1950s, pressurised Metered Dose Inhalers (pMDIs) are one of the most
commonly used methods of delivering drug dosages to the lung[1].

As aerosol canisters which contain drug and liquid propellant (as well as surfactant and other
excipients), drugs were at first delivered suspended or dissolved in ChloroFluoroCarbons (CFCs)[1].
Such propellants have now been replaced with HydroFluoroAlkenes in compliance with the Montreal
Protocol on Substances that Deplete the Ozone Layer[2].

Despite the fact that pressurised Metered Dose Inhalers have been around for over 50 years however,
a notable and recurring disadvantage (or perhaps issue) is that patients often have problems
coordinating actuation and inhalation of the device[1], leading to poor patient adherence and
consequently, sub-therapeutic effects[3]. This is notably seen in children and the elderly who both
show dependence on the caregiver[4]. The use of a spacer is thought to reduce these issues (as well as
electronic inhaler devices that are able to mechanically trigger actuations). Unfortunately, such
electronic devices are still quite rare due to their high cost[5].

On that note, cheaper, generic inhaled products are now available on the market alongside established
branded products for the same indication. As either of the two products (such as the Ventolin ®
inhaler and the generic Salbutamol inhaler used in this study) can be prescribed, and with many
patients preferring the cheaper product, bioequivalence testing must take place in order to prove that
the generic inhaler will have the same safety profile and deliver the same therapeutic outcome as the
branded inhaler.

This study therefore addresses two questions. Does the generic inhaler show bioequivalence to the
branded inhaler, and is therefore able to deliver the same therapeutic outcome? What effect does a
spacer have for both inhalers, on the Respirable Fraction (percentage of drug that reaches the target
site)?

Hypothetically, the data derived from this experiment should show evidence that the two inhalers are
bioequivalent and the use of a spacer should demonstrate increased efficacy in drug delivery to the
site of action; the lower airways.

1
Hyun Jung Lee (1310438) RMS 9

MATERIALS AND METHODS

The two inhalers used (and tested) in this experiment were the Ventolin Evohaler ® and the CFC-
Free Salbutamol Inhaler. As pressurised metered-dose inhalers, both contained Salbutamol Sulphate
as its active ingredient (with a dose of 100 µg per actuation), anhydrous ethanol and HFA-134a
(propellant). The Ventolin® was supplied by GlaxoSmithKline and the generic Salbutamol inhaler
was supplied by IVAX pharmaceutics. 

Salbutamol Calibration Curve

Salbutamol basestock solution (100 µg/mL) was diluted to concentrations 80, 60, 40, 20, 10 µg/mL by
addition of Methanol:Water (1:1), resulting in six volumetric flasks of total volume 10 mL of varying
concentrations. After ensuring the contents of each flask was mixed thoroughly, each solution (with
their respective concentrations) was transferred into a Quartz UV Cuvette for absorbance
measurements by a UV Spectrometer (276nm using a cuvette of 1cm path length). These absorbance
values were plotted against concentration (µg/mL), producing a Calibration Curve and its Linear
Equation. The Linear Equation was used to calculate Salbutamol concentrations and the Coefficient of
Determination (R2) allowed evaluation of the extent to which the regression line on the graph fit the
data points in the table.

Total Emitted Dose

Two 100 mL beakers were each filled with 35 mL of solvent (Methanol:Water 1:1). A single metal
disc (with a hole for inhaler valve stems) was placed inside each beaker. Both inhalers (Ventolin ®
and Generic) were primed by being shaken and discharged (twice) in the sink. Actuator sleeves were
removed from both inhalers and the valve stems were placed inside the holes of the metal disc, ready
for actuating. Each inhaler was actuated fully ten times, with spaces to allow time between actuations.
The two 35 mL solutions were then transferred to a 50 mL volumetric flask and made up to 50 mL by
addition of solvent. This solution was then transferred to a Quartz UV Cuvette for absorbance
measurements by a UV Spectrometer (276nm using a cuvette of 1cm path length). The absorbance
values from this experiment were used to calculate the mass of Salbutamol per actuation (in
conjunction with the Linear Equation derived from the Salbutamol Calibration Curve).

Testing Aerosol Performance with the Twin-Stage Impinger

The Twin-Stage Impinger was set up according to the standard British Pharmacopeia method
(Volume IV Appendices: Appendix XII). A total of four tests were carried out in the same set-up, the
only changes being the type of inhaler and the presence (or absence) of a spacer.

1. Ventolin ® Inhaler.
2. Generic Salbutamol Inhaler.
3. Ventolin ® Inhaler and Spacer.
4. Generic Salbutamol Inhaler and Spacer.

For each test, the inhaler was firstly primed (by being shaken and discharged) and the Twin-Stage
Impinger pump was switched on. Then (with or without spacer), the inhaler was attached to the
mouthpiece and actuated for a total of twenty times. The solutions in separate components of the
Twin-Stage Impinger (Throat Piece B, Upper Flask D, Lower Flask G) were collected and transferred
into separate volumetric flasks (50 mL, 20 mL, 50 mL respectively) and made up to their flask
volumes by addition of solvent (Methanol:Water 1:1). The solutions were then transferred to Quartz
UV Cuvettes for absorbance measurements by a UV Spectrometer (276nm using a cuvette of 1cm

2
Hyun Jung Lee (1310438) RMS 9

path length). The absorbance values from this experiment were used to calculate the percentage of
drug deposited in each compartment of the Twin-Stage Impinger (the percentage drug in the lower
flask being the Respirable Fraction percentage).

Data for “Total Emitted Dose” and “% Labelled Dose of Salbutamol” was presented as Mean ±
Standard Deviation form (n=3 evaluations) for each of the three groups in the laboratory.

How the Data was Calculated

1. Calculating Concentration of Salbutamol (µg/mL):

For Group 1 using Linear Equation 1:

y = 0.0060x + 0.0086

x = (y - 0.0086)/0.0060

y = absorbance, so substituting an example absorbance value for Ventolin Group 1,

x = (0.220 – 0.0086)/0.0060 = 35.2 µg/mL

For Group 2 using Linear Equation 2:

y = 0.0062x + 0.0028

x = (y - 0.0028)/0.0062

y = absorbance, so substituting an example absorbance value for Ventolin Group 2,

x = (0.125 – 0.0028)/0.0062 = 19.7 µg/mL

2. Calculating Mass of Salbutamol per Actuation (µg):

Concentration of Salbutamol ( µg /mL ) ×Volume (mL)

Number of Actuations

Volume = 50mL (or 20mL for Upper Chamber)

Actuations = 10 (for TED) or 20 (for TSI)

Example: Concentration (eg 35.2 µg/mL x 50mL)/10 Actuations = 176.0 µg

3. Calculating Total Emitted Dose (% of Label Dose):

Amount of Salbutamol recovered ¿ the TSI (µg ) ¿ ×100

Labelled Dose of Salbutamol (µg)

Example: (176.0 µg/100) x 100 = 176.0%

4. Calculating Respirable Fraction (%):

Amount of Salbutamol∈lower chamber of TSI (µg)
×100
Labelled Dose of Salbutamol( µg)

Example: 17.900/100 x 100 = 17.9%

3
Hyun Jung Lee (1310438) RMS 9

RESULTS

Salbutamol Calibration Curve

0.7

0.6

0.5
Absorbance at 276nm

0.4

0.3

0.2

0.1

0
0 20 40 60 80 100 120

Salbutamol Concentration (µg/mL)

Absorbance value at 276nm (3 d.p.) Group 1
Linear (Absorbance value at 276nm (3 d.p.)
Group 1)
Absorbance value at 276nm (3 d.p.) Group 2
Linear (Absorbance value at 276nm (3 d.p.)
Group 2)
Absorbance value at 276nm (3 d.p.) Group 3

Figure 1: Three Salbutamol Calibration Curves showing absorbance values (measured at 276nm)
of solutions of known drug concentration (μg/mL), later used to calculate unknown concentrations
of Salbutamol in a branded and generic inhaler

Group 1 Group 2 Group 3

Linear Equation y=0.0060x + 0.0086 y=0.0062x + 0.0028 y=0.0063x - 0.0101
Coefficient of 0.9998 0.9974 0.9999
Determination (R2)

Table 1: Salbutamol Base Calibration Curve Equations from each group, with their respective
coefficients

Three calibration curves were plotted (one for each group) to enable conversion of absorbance values
into concentration values of Salbutamol in a branded and generic inhaler. Hypothetically, if the
method was followed correctly, all three calibration curves should overlap with the same linear
equation for all three curves. Figure one from the experiment indicates that the three calibration
curves do not completely overlap, showing slight variations. Possible explanations include incorrect
measurements of volumes (for example, going over the 10mL mark when filling up the flask with
solvent), improper mixing of volumetric flask contents, improper washing of the Quarts UV cuvette
and the outside of the cuvette not being clean and dry. However, the variations are not too extreme,
with the coefficient of determination (R2) values for all three groups being higher than 0.999 and close
to 1 and the y-intercepts being close to 0. This indicates that the errors were most probably down to
different people from different groups carrying out the experiment, giving room for slight variations
in data.

4
Hyun Jung Lee (1310438) RMS 9

Total Emitted Dose

200
180

Mean Total Emitted Dose as

160
140

% of Label Dose
120
100
80
60
40
20
0
Ventolin® Generic

Figure 2: A Comparison of the Mean Total Emitted Dose Values as a Percentage of Label Dose
(%) for Ventolin and the generic pMDI Device. Values represent the mean ± standard deviation of
n=3 batches

The mean Total Emitted Dose should be the same for both the branded and generic inhalers if the two
are said to be bioequivalent. However, our data did not show evidence of bioequivalence as the
generic inhaler emitter larger amounts of drug (176.0%) than the branded inhaler (128.0%). Both of
these results are larger than expected and henceforth, not in compliance with the British
Pharmacopoeia requirement which states that the “mass balance should be no less than 75% and no
greater than 125% of the label dose”[6].

We can attribute this unexpected result to human errors, many of which were possible to make
throughout the experiment. For example, inhalers may not have been primed properly before
actuation. As pressurised Metered Dose Inhalers contain suspensions, flocculation may occur when
the inhaler is at rest. This can lead to clogging of the valve system, preventing the full dose of the
actuation from being emitted into the solvent. On the other hand, excessive shaking can lead to
foaming[7]. Inhalers must be correctly shaken to avoid these problems, yet the force of the shaking
will have varied between people and between groups. The force of the actuation may also not have
been consistent (for instance, the first actuation being weaker than the last). This too, will have varied
between inhalers, between people and between groups. Furthermore, it was possible for students to
make errors regarding volumes, (for example, measuring 35mL of solvent incorrectly or going over
the 50mL when making up the volumetric flask with solvent). These potential errors would have
contributed to inaccurate absorbance values, leading to inaccurate Mean Total Emitted Dose values.

5
Hyun Jung Lee (1310438) RMS 9

Testing Aerosol Performance with the Twin-Stage Impinger

Inhaler ± Spacer Percentage Respirable Fraction

Mean ± Standard Deviation

Ventolin® Inhaler 43.3 ± 22.5

Generic Salbutamol Inhaler 13.7 ± 9.9
Ventolin® Inhaler and Spacer 24.6 ± 3.8
Generic Salbutamol Inhaler and Spacer 53.5 ± 6.7
Table 2: The Mean ± Standard Deviation Values for the Respirable Fraction of Ventolin and the
Generic pMDI Both Without and With a Spacer. Values represent the mean ± standard deviation
of n=3 batches

Literature values from the Food and Drug Administration state that approximately ten to fifteen
percent of the dose for oral inhalation aerosols reaches the target site (the rest is trapped in the mouth,
upper airways and is swallowed or absorbed by the gastrointestinal tract)[8]. Three of the values in
Table 2 (with the exception of the Generic Salbutamol Inhaler without a spacer) were higher than the
FDA expected outcome of ten to fifteen percent. After careful analysis of the raw data, it was clear
that no patterns could be identified to suggest that data analysis errors were made. (For future
experiments, it would be wise to validate the experiment with a standard – with a known deposition
profile - to confirm whether the TSI was set up correctly according to the BP standards).

The first is that patients using inhaler devices already have impaired breathing, meaning that the pump
used as part of the Twin-Stage Impinger to replicate the air stream of a patient is more likely to collect
a larger amount of drug than a patient with breathing difficulties can. The pump also does not take
exhalation into account; a portion of drug particles are sometimes exhaled immediately upon
inhalation[9]. Henceforth, more of the drug may reach the lower chamber in in vitro conditions than
in vivo.

The second possible (and more likely) explanation is that this experiment particularly (than the
previous two experiments) had more room for error. These include: incorrect measurements of solvent
volume, incorrect assembly of the Twin-Stage Impinger (leaving gaps for potential escape of drug
particles), not fully connecting the pump to the Twin-Stage Impinger, insufficient priming of the
inhaler, only partial attachment of the inhaler to the mouthpiece, improper attachments of the inhaler
to the spacer, insufficient shaking of the inhaler between actuations, incorrect counting of actuations,
transfer of solutions to the wrong volumetric flask, going over the 20mL/50mL mark with solvent,
and incomplete washing and drying of the Twin-Stage Impinger by previous groups, leaving drug
particles to collect on the apparatus.

These factors all could have contributed to our data which shows an excessive, overestimation of the
Respirable Fraction.

In regards to bioequivalence, our results for the Respirable Fraction of the branded and generic
inhalers are not similar (Ventolin: 43.3 Generic: 13.7 without spacers, Ventolin: 24.6 Generic: 53.5
with spacers), which they should be if they are said to be truly bioequivalent. As both products are
established products out on the market however, the large differences must be accounted to the errors
mentioned above, which is proved by the standard deviation values and can be seen visually by the
large error bars marked on the graphs below.

6
Hyun Jung Lee (1310438) RMS 9

70

Percentage Drug Deposition (%) in

the Throat Piece, Upper Chamber
60

50

and Lower Chamber

40

30

20

10

0
Actuator/Throat piece Upper Chamber Lower Chamber
(± spacer)

Ventolin® Generic

Figure 3: Bar Graph Comparing the Drug Deposition in the Throat, Upper and Lower Flasks for
Ventolin and the Generic pMDI WITHOUT a Spacer

70
Percentage Drug Deposition (%) in the
Throat Piece, Upper Chamber and

60

50
Lower Chamber

40

30

20

10

0
Actuator/Throat piece Upper Chamber Lower Chamber
(± spacer)

Ventolin® Generic

Figure 4: Bar Graph Comparing the Drug Deposition in the Throat, Upper and Lower Flasks for
Ventolin and the Generic pMDI WITH a Spacer

For both the branded and generic inhalers, the use of a spacer notably decreased the mean dose of
drug depositing in the throat piece and upper chamber, proving that the presence of a spacer had
positive effects on reducing the deposition of drug particles on parts of the respiratory tract which are
not the intended target site. This was further proved when looking at the results of the generic
Salbutamol inhaler, whereby our results showed that the use of a spacer dramatically increased the
Respirable Fraction from 13.7% to 53.5%. However, this could not be said for Ventolin, whereby the
Respirable Fraction decreased from 43.3% to 24.6%. As Ventolin is a branded product already out on
the market, this unexpected result is most probably due to human error (explained above), which is
further reinforced by the largest standard deviation value (22.5). This large value suggests high
variability between groups and therefore the 43.3% for the branded inhaler is not representative of the
true deposition profile of Salbutamol in the Ventolin ® inhaler.

7
Hyun Jung Lee (1310438) RMS 9

DISCUSSION

Orally inhaled products are tested on their bioequivalence in vitro with the use of a Twin-Stage
Impinger, whose various stages represent specific sections of the respiratory tract (throat, upper
airways and lower airways)[10]. The purpose of bioequivalence testing orally inhaled products is not
necessarily to draw comparisons between branded and generic products, but to show evidence of their
equivalence[11]. Thus, if two orally inhaled drugs are said to be bioequivalent, under test conditions
they must show similar bioavailabilities and therefore (according to the Food and Drug
Administration) no “significant difference in the rate and extent to which the active ingredient…
becomes available at the site of drug action when administered at the same molar dose…” [12] For
orally inhaled drugs, this means that the Total Emitted Dose and the percentage of drug collected in
the lower chamber of the TSI (representative of the lower airways - the drug target site) should be
similar, if not the same. Bioequivalence testing is crucial in the pharmaceutical industry as patients
may be prescribed either of the two inhalers (branded or generic) yet the therapeutic effect is expected
to be no different.

The Respirable Fraction is defined as the fraction of the Total Emitted Dose that reaches the lower
chamber of the TSI which models the lower airways; the local site of action of the drug[13]. In the
lower airways, Salbutamol relaxes smooth muscle and causes bronchodilation by binding to β2-
adrenergic receptors[14]. The Respirable Fraction is therefore critical in comparing two inhalers when
regarding bioequivalence of orally inhaled products, as most bioequivalence definitions consider
systemic absorption, which is not as relevant for inhaled drugs. This is in agreement with the
European Medicines Agency Guideline, which associates fine particle mass of the drug reaching the
lung with efficacy and safety of the drug product[15].

The efficacy of inhalers can be increased by the use of spacers, which reduce deposition of large
droplets in parts of the respiratory tract which are not the intended target sites of the drug (throat and
upper airways), allowing more drug to reach the site of action (the lower airways). This can prevent
side effects that tend to occur as a result of the drug depositing in the throat and airways[16]. Spacers
also prevent inhalation/actuation coordination problems experienced by the patient. This is because
the spacer acts as a reservoir for drug particles before inhalation by the patient occurs, so that droplet
velocity decreases, large particles are eliminated and the propellant is given more time to evaporate,
ensuring that inhalation/actuation coordination is no longer necessary[1].

Our data did not show evidence of bioequivalence of the generic device compared to Ventolin ®. In
regards to the Total Emitted Dose values, the generic inhaler emitter larger amounts of drug than the
branded inhaler. The Respirable Fraction values were also very different (as mentioned in the results).
This large gap between the values can be explained when looking at standard deviations for all values,
which indicate poor reliability and reproducibility of data, thus inaccurate results.

(500 Words)

CONCLUSION

Whilst our data did not provide evidence of the branded and generic inhaler having bioequivalence
nor did it fully show the therapeutic advantages of a spacer, as two established products out on the
market (Ventolin® and CFC-Free Salbutamol), we must assume that the data produced in our
experiment is inaccurate.

8
Hyun Jung Lee (1310438) RMS 9

Overall, inhaler performance depends on many factors, for instance the breathing profile of the
patient, the patient’s inhaler technique, the force of the actuation, the aerodynamic diameter and initial
velocity of the aerosol particles and the use of a spacer. To counteract the variety of such factors
which may lead to sub-therapeutic effects, pressurised Metered Dose Inhalers usually have doses of
100mcg, which is much higher than what eventually reaches the site of action.

REFERENCES

[1] Michael E. Aulton and Kevin M. G. Taylor. Aulton’s Pharmaceutics, The Design and Manufacture
of Medicines by 4th Edition. Elsevier Ltd. 2013.

[2] G. J. M. Velders, A. R. Ravishankara, M. K. Miller, M. J. Molina, J. Alcamo, J. S. Daniel, D. W.

Fahey, S. A. Montzka, S. Reimann. Preserving Montreal Protocol Climate Benefits by Limiting
HFCs. Science, 2012; 335 (6071): 922

[3] http://www.clinicalcorrelations.org/?p=5129 (Accessed 08/03/2015 21:43)

[4] Kimberly P. Raymond; Barbara H. Fiese;Marcia A. Winter; Andrea Knestel; Robin S. Everhart.

Helpful Hints: Caregiver-Generated Asthma Management Strategies and Their Relation to Pediatric
Asthma Symptoms and Quality of Life Oxford Journals 2011 Volume 37, Issue 4 Pp. 414-423

[5] John Wright; David Brocklebank; Felix Ram. Inhaler devices for the management of asthma and
COPD. University of York, Volume 8, Number 1, 2003

[6] The British Pharmacopoeia Online 2015. Available from: http://www.pharmacopoeia.co.uk/

[7] Document from the European Medicines Agency: Doc Ref.: EMEA/CHMP/QWP/49313/2005
Corr London, 21 June 2006. Available from:
http://www.ema.europa.eu/docs/en_GB/document_library/Scientific_guideline/2009/09/
WC500003568.pdf (Accessed 08/03/2015 21:43)

[8] Document from the Food and Drug Administration 1998. Available from:
http://www.fda.gov/ohrms/dockets/ac/00/backgrd/3634b1c_Sectiond.pdf (Accessed 08/03/2015
21:43)

[9] European Pharmacopoeia. Available from::

http://lib.njutcm.edu.cn/yaodian/ep/EP5.0/02_methods_of_analysis/2.9.__pharmaceutical_technical_p
rocedures/2.9.18.%20Preparations%20for%20inhalation%20%20aerodynamic%20assessment%20of
%20fine%20particles.pdf (Accessed 08/03/2015 21:43)

[10] Christina Spyridoi; Mark H Saunders; Simon Gaisford. Thermal Characterisation of BDP From
Dry-Powder Inhalers. University College of London Research Poster 2010, tcm18-230171.

[11] Stephen H. Curry, Robin Whelpton. Drug disposition and pharmacokinetics: from principles to

applications. Chichester, West Sussex ; Hoboken, N.J. : Wiley-Blackwell, 2011.

[12] Document from the Food and Drug Administration 2003. Available from:
http://www.fda.gov/downloads/Drugs/Guidances/ucm070124.pdf (Accessed 08/03/2015 21:43)

[13] Nieboer, E., Thomassen, Y., Chashchin, V., Odland, J.O. 2005. Occupational Exposure
Assessment of Metals. JEM 7: 411-415.

[14] http://salbutamol.org/ (Accessed 08/03/2015 21:43)

9
Hyun Jung Lee (1310438) RMS 9

[15] Document from the European Medicines Agency. Available from:

http://www.ema.europa.eu/docs/en_GB/document_library/Scientific_guideline/2009/09/
WC500003568.pdf (Accessed 08/03/2015 21:43)

[16] http://www.asthma.org.uk/knowledge-bank-treatment-and-medicines-spacers (Accessed

08/03/2015 21:43)

APPENDIX

Results Table 1: Group Members

Group Number Subgroup A Subgroup B Subgroup C Subgroup D

1 Ming-Yiu Chong Hyun Jung Lee Harpreet Kaur Huma Wardak
Spike Ibeji Samira Virani Meera Mamtora Ali Shabaan
Rola Ibrahim Zainab Sumar

Results Table 2: Absorbance Values of Salbutamol Base Calibration Curve Dilutions

Concentration Absorbance Values at 276nm (3 d.p.)

(μg/mL) Group I Group 2 Group 3
100 0.610 0.630 0.617
80 0.487 0.500 0.490
60 0.365 0.362 0.362
40 0.245 0.235 0.238
20 0.128 0.129 0.116
10 0.072 0.077 0.054

Results Table 3: Salbutamol Base Calibration Curve Equations

Group 1 Group 2 Group 3

Linear Equation y=0.0060x + 0.0086 y=0.0062x + 0.0028 y=0.0063x - 0.0101
Coefficient of 0.9998 0.9974 0.9999
Determination (R2)

Results Table 4: Absorbance Values for Total Emitted Dose Experiment

Sample Absorbance Values at 276nm (3 d.p.)

Group 1 Group 2 Group 3
Ventolin® 0.220 0.125 0.128
Generic 0.230 0.238 0.184

Results Table 5: Calculations for Total Emitted Dose Experiment

Group 1 Group 2 Group 3

Concentration Ventolin® 35.2 19.7 21.9
Salbutamol
(μg/mL) Generic 36.9 37.9 30.8

Mass Salbutamol Ventolin® 176.0 98.5 109.5

per actuation

10
Hyun Jung Lee (1310438) RMS 9

(μg) Generic 184.5 189.5 154.0

Total Emitted Ventolin® 176.0 98.5 109.5

Dose (% of
Label Dose) Generic 184.5 189.5 154.0
Mean of Total Ventolin® 128.0
Emitted Dose Generic 176.0
(%)
Standard Ventolin® 41.9
Deviation Generic 19.2

Results Table 6: Salbutamol Deposition Profile in Different Inhalers ± Spacers (Absorbance

Values)

MDI± Spacer Class Group Absorbance Values at 276nm (3 d.p.)

Actuator/throat Upper Chamber Lower Chamber
±
piece ( spacer)
1 Ventolin® Group 1 0.168 0.056 0.051
Group 2 0.188 0.084 0.147
Group 3 0.082 0.072 0.113
2 Ventolin® + Group 1 0.146 0.007 0.063
Spacer Group 2 0.014 0.017 0.071
Group 3 0.016 0.009 0.043
3 Generic Group 1 0.074 0.040 0.0014
Group 2 0.048 0.024 0.053
Group 3 0.018 0.022 0.031
4 Generic and Group 1 0.005 0.013 0.135
Spacer Group 2 0.005 0.015 0.146
Group 3 0.009 0.031 0.103

Results Table 7: Calculations for Salbutamol Concentrations (μg/mL)

MDI± Spacer Class Group Concentration of Salbutamol Base (μg/mL) (3 d.p.)

Throat Piece Upper Chamber Lower Chamber
1 Ventolin® Group 1 26.600 8.000 7.160
Group 2 29.870 13.100 23.258
Group 3 15.300 13.600 20.500
2 Ventolin® + Group 1 23.000 -0.160* 9.160
Spacer Group 2 1.806 2.290 11.000
Group 3 4.300 3.160 8.830
3 Generic Group 1 11.000 5.300 1.000
Group 2 7.290 3.419 8.097
Group 3 4.600 5.300 6.830
4 Generic and Group 1 -0.500* 0.830 21.160
Spacer Group 2 0.355 1.968 23.097
Group 3 3.160 6.830 18.830
*Minus results possibly due to contaminants in the spacer device which reduce UV absorbance

Results Table 8: Calculations for Salbutamol Mass per Actuation (μg)

11
Hyun Jung Lee (1310438) RMS 9

MDI± Spacer Class Group Mass of Salbutamol per Actuation (μg) (3 d.p.)
Actuator/throat Upper Chamber Lower Chamber
piece (± spacer)
1 Ventolin® Group 1 66.500 8.000 17.900
Group 2 74.675 13.100 58.145
Group 3 38.250 13.600 51.250
2 Ventolin® + Group 1 57.500 -0.160 22.900
Spacer Group 2 4.515 2.290 27.500
Group 3 10.750 3.160 22.075
3 Generic Group 1 27.500 5.300 2.500
Group 2 18.225 3.419 20.242
Group 3 11.500 5.300 17.075
4 Generic and Group 1 -1.250 0.830 52.900
Spacer Group 2 0.888 1.968 57.743
Group 3 7.900 6.830 47.075

Results Table 9: Calculations for % Labelled Dose of Salbutamol

MDI± Spacer Class Group % Labelled Dose of Salbutamol (1 d.p.)

Actuator/throat Upper Chamber Lower Chamber
piece (± spacer)
1 Ventolin® Group 1 66.5 8.0 17.9
Group 2 77.9 13.1 60.8
Group 3 38.3 13.6 51.3
Mean 60.9 11.6 43.3
SD 20.4 3.1 22.5
2 Ventolin® + Group 1 57.5 -0.2 22.9
Spacer Group 2 5.4 2.3 29.0
Group 3 10.8 3.2 22.1
Mean 24.6 1.8 24.6
SD 28.6 1.8 3.8
3 Generic Group 1 27.5 5.3 2.5
Group 2 19.6 3.4 21.5
Group 3 11.5 5.3 17.1
Mean 19.5 4.7 13.7
SD 8.0 1.1 9.9
4 Generic and Group 1 -1.3 0.8 52.9
Spacer Group 2 1.5 2.0 60.4
Group 3 7.9 6.8 47.1
Mean 2.7 3.2 53.5
SD 4.7 3.2 6.7

12