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Review of Taste Evaluation Technologies for The

Development of Oral Paediatric Dosage Forms
Dr. Abeer Ahmed-Mohammed
Dr. Catherine Tuleu
Dr. Terry Ernest

Project co-funded by
Accelerating Paediatric Formulation Development
through Smart Design and Predictive Science.

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Accelerating paediatric formulation development through smart

design and predictive science
Workstream 1
Aim: to recommend a best practice for assessment of taste to support the
development of age appropriate dosage forms.

We conducted a systematic literature review of in vitro and in vivo taste

assessment tools (non-human).
Purpose: acquire, combine, and shape scientific techniques used in
taste evaluation.

Project co-funded by

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Why taste of medicine is important?

The taste of medicines have a huge impact on patients acceptability and

compliance especially in the paediatric population.

Most drugs have bitter taste which poses a challenge for development of oral
pharmaceutical dosage forms.

Taste assessment should be conducted early in development which drives a need

for taste masking strategies.

The most common taste assessment method is human taste panel but it has
several drawbacks such as; (I) individuals taste perception variability, (II) ethical
and safety and (III) it is difficult for children to give reliable taste perception
statement (adults only).

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Methods for taste assessment (non-human) of medicine systematic review

In vitro
1.

Electronic tongues.

2. Release/dissolution tests.
Other in vitro methods: Cell based assays.

In vivo
1. Animal behavioural model: mainly rodents
Two bottles preference test
Brief access taste aversion test (BATA)
2. Electrophysiological methods
Other models: Drosophilia and Zebra fish

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Scoring of most commonly used tools for taste assessment of medicines

Animal model
Criterion
Usability
SOP use easy for trained technical
staff or experienced user
Needs a licence or REC approval
Speed (time from setup to data
collection)
Ability to screen pure drug/product
Drug, excipients
Formulated product (ideally other than
solution e.g. suspension)
Evidence of correlation to in vivo
data
Potential to validate
Specificity
Linearity
Accuracy
Precision
Range-quantification limit-detection
limit
Costs
Initial investment (platform)
Running costs
Sum of scores

Electronic tongue

BATA

Electrophysiological

Two
bottles

Astree

Insent

1
2

1
1

1
1

5
3

5
3

5
3

4
0

4
3

5
3

5
3

5
5
5
4
3

5
4
4
0
3

5
1
3
0
3

5
4
0
1
1

5
4
0
2
1

2
4
46

1
1
26

2
4
33

2
2
39

2
3
42

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In vitro: Taste sensing systems

electronic tongues measure changes in electrical potential while testing

liquid samples
Two most commonly used for pharma
(i) SA402B/ TS-5000z (Insent Inc, Japan):
each sensor is assigned to specific taste with partial cross selectivity.
(ii) -Astree e-tongue (Alpha M.O.S, France):
sensors are cross selective.
Reliable taste data can be achieved when there is a linear relationship
between log concentration of the tastant and sensor signal.

Insent Electronic tongue

Alpha-Astree Electronic tongue

Chen, Z., et al., Methods, 2012. 4(3): p. 599-608..

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In vivo: Brief access taste aversion test (BATA)

Presentation of bitter solutions to mildly water deprived rodents (22 h) for

very brief period of time (8 seconds).
The frequency of drinking is quantified vs water (number of licks).
Licks are recorded by a lickometer connected to computer.
Bitterness intensity inversely proportional to number licks
Limitation: new drugs with no toxicity data.

Soto, J., Sheng, Y., Standing, J., Orlu Gul, M. & Tuleu, C. Development of a model for robust and exploratory analysis of
the rodent brief-access taste aversion data. Eur. J. Pharm. Biopharm. (2015)

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Conclusion
The most common in vitro taste evaluation method is electronic taste
sensor mainly either
(i) SA402B/ TS-5000z (Insent Inc)
(ii) -Astree e-sensor (Alpha M.O.S).

The Animal behavioural test (mainly rodents and drosophilia flies) is

promising tool for taste assessment of medicine.

Novel cell based taste assessment methods will be extensively

explored (Aston University).
The suitability for any of the methods to be used in replace of or in
conjunction with human taste panel tests will be determined by the
IUK project.

Project co-funded by
Accelerating Paediatric Formulation Development
through Smart Design and Predictive Science.

UCL SCHOOL OF PHARMACY

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Accelerating Paediatric Formulation

Development Through Smart Design and
Predictive Science Cell based taste
assessment assay

Afzal Mohammed
Craig Russell

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Cell based taste assessment models

Cell based taste

assessment

Excised tissue
Eg: bovine tongue

Eukaryotes

Prokaryotes
Eg: amoeba

Primary cells
Eg: rodents, human
cells

Bioengineered cells
Eg: human
embryonic kidney
cells (HEK 293)

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Cell based taste assessment models bovine taste receptor cells

Epithelial
Cells

Outer Taste
Pore

Microvill
i

Bovine tongue
Support Cells (Type I, dark
cells (50%))
Sensory Cells (Type II and III,
light cells (25%))
Basal Cells
(Type IV)
Synapse

Nerve

Schematic representation of various different cells within a taste bud

Contain large number of

taste papillae
Isolation of taste buds is
convenient due to large
size
Pharmacology of taste
response similar to
human
Ethical - by product from
the beef industry

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Light microscopy images of bovine taste receptor cells

- primary culture

2:21
following
first media
Day 1:14
hours after
seeding
7 24
change.

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Isolation and culture salient features

No enzymatic digestion step during the extraction stage

Once the cells had been grown in the 24 well plate, it was
possible to transfer the cells to vessels which had not
been coated with collagen and still observe growth
unchanged from that observed on coated glass and
plastic ware
Cells have been successfully frozen down providing a stock
of cells for future use
The frozen cells have been successfully revived
The revived cells have the same morphology and also have
been shown to behave the same as the primary cells
obtained from fresh culture.

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Confocal Imaging for the Identification of TRC markers Gustducin and PLC-B2

As a secondary means of confirming the cultured cells are

certainly TRCs, confocal microscopy was used to identify
two proteins involved in taste transduction - Gustducin and
PLC-B2.

The cells were again grown on coverslips before they were

stained with primary and secondary antibodies.
The primary antibody solutions were Rabbit polyclonal
Ggust (I-20) and Rabbit polyclonal PLC-2 (Q-15).
The secondary antibodies were Goat anti-rabbit IgG Alexa
633 and Goat anti-rabbit IgG Alexa 488

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Confocal Imaging for the Identification of TRC

markers Gustducin and PLC-B2

TRCs stained with Rabbit polyclonal

Ggust (I-20) followed by Goat antirabbit IgG Alexa 488 showing presence
of Gustducin.

TRCs stained with Rabbit polyclonal

PLC-2 (Q-15) followed by Goat antirabbit IgG Alexa 633 showing presence
of PLC-B2.

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Intracellular calcium response to taste stimuli

Cells Before Stimuli

Cells After Stimuli

Exemplar response for 2mM Denatonium (model tastant)

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Intracellular calcium response to taste stimuli

The images presented are passage 4 cells and are representative of

images also gathered at passage 2 and passage 7.

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Intracellular calcium response to taste stimuli

Intracellular Calcium Response of BTRC's to
2mM Denatonium

1.25

1.2

Fluroesence Intensity Ratio

Fluroesence Intensity Ratio

1.3

Intracellular Calcium Response of BTRC's to

2mM Sucralose
1.10

1.05

1.15
Calcium Response

1.1
1.05

Calcium

1.00

1
0.95

0.95

0.9
0.85
0.8
0

100

200

300
Field Time (s)

400

500

0.90
0

100

200
300
Field Time (s)

400

500

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Conclusions

Bovine taste receptor cell cultures were established which maintain

their morphology and functionality up to at least passage 7.
Characterisation of BTRCs confirmed the presence of gustducin and
PLC 2

Calcium imaging using fluorescence microscopy confirmed the

differential response to tastants

Ongoing work is focussed at :

1. correlating data from in vitro/in vivo tools with human taste
panel results
2. evaluation of taste masking efficacy