International Journal of Pharmaceutics 447 (2013) 199203

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International Journal of Pharmaceutics

journal homepage: www.elsevier.com/locate/ijpharm

An in vitro model for the evaluation of the adhesion of solid oral dosage forms to
the oesophagus
John D. Smart a, , Sian Dunkley b , John Tsibouklis b , Simon Young c
a
b
c

School of Pharmacy and Biomolecular Sciences, University of Brighton, Brighton, BN2 4GJ, United Kingdom
School of Pharmacy and Biomedical University Sciences, University of Portsmouth, Portsmouth, United Kingdom
School of Medical Sciences, RMIT University, PO Box 71, Bundoora, Victoria 3083, Australia

a r t i c l e

i n f o

Article history:
Received 15 October 2012
Received in revised form 5 February 2013
Accepted 6 February 2013
Available online 1 March 2013
Keywords:
Oesophageal adhesion
Bioadhesion
Mucoadhesion
Non-adhesive coatings

a b s t r a c t
Adhesion of solid oral dosage forms to the oesophagus can be a disadvantage when delivering drugs that
may cause oesophageal damage, or can be an advantage when developing localised therapies for this
region. In this study, apparatus to investigate coatings that may inuence oesophageal retention was
developed and evaluated. The apparatus incorporated a section of porcine oesophageal mucosa held at
by the application of a gentle vacuum and kept moist by the application of a simulated saliva solution. The
resistance to the application of more physiologically relevant shear stresses was evaluated. Using a range
of materials it was found that differences in oesophageal adhesion could be identied. Materials like
sodium alginate were highly adhesive and had a tendency to re-adhere while parafn waxes showed no
adhesion. The rapid loss of the polymer coat from the surface for water swellable materials was identied
as an issue.
2013 Elsevier B.V. All rights reserved.

1. Introduction
The adhesion of solid formulations to the oesophagus during
swallowing has been widely implicated in medication-induced
injury to this organ. Adherence during swallowing results in a high
local drug concentration within the oesophagus, which may lead to
oesophageal damage when the drug is irritant. Formulations containing emeporonium bromide, apple cider vinegar, alendronate
sodium, tetracycline and potassium chloride are implicated in such
damage (Hill et al., 2005; Jaspersen, 2000; Ueda et al., 2011). Moreover, formulation adhesion within the oesophagus usually leads to
a delay in the onset of action of the drug, which may profoundly
affect both bioavailability and pharmacokinetics. Formulation size,
shape and surface characteristics (coating) have been identied as
factors that affect dosage-form adhesion in the oesophagus during
swallowing (Channer and Virjee, 1985; Marvola et al., 1982; Perkins
et al., 2001). However, adhesion to the oesophagus could also be
advantageous, for example in locating a formulation for the treatment of gastrooesphageal reux disease (Batchelor et al., 2002),
pain and inammation (Mako et al., 2009) or for delivering diagnostic agents (Collaud et al., 2007). The overall aim of this study is to
develop an in vitro apparatus to investigate both putative adhesive

Corresponding author. Tel.: +44 1273 642091; fax: +44 1273 6420.
E-mail addresses: john.smart@brighton.ac.uk, the smart family@btinternet.com
(J.D. Smart).
0378-5173/$ see front matter 2013 Elsevier B.V. All rights reserved.
http://dx.doi.org/10.1016/j.ijpharm.2013.02.017

and non-adhesive coating materials, for use in the development of

solid oral dosage forms.
Oesphageal retention has been evaluated in vivo using gamma
scintigraphy (Drake et al., 2002; Perkins et al., 2006; Sakkinen et al.,
2004) and some in vitro models have also been developed, notably
for gels and liquids (e.g. Batchelor et al., 2002). An oesophageal
model where a formulation was placed into the section of pigs
oesophagus was found not to be a good model for in vivo transit of
tablets (McCargar et al., 2001). This may be due to the mucosa being
suspended within the apparatus and not providing a uniformly at
surface for this evaluation. The adhesion of lm coated tablets to
oesophageal mucosa has also been investigated using a tensile like
testing apparatus (Al-Dujaili et al., 1986a,b).
In this work, a novel in vitro apparatus modelling the adhesion of
solid formulations to oesophageal tissues has been developed and
optimised, with a view to its use in the evaluation of coatings for
solid oral dosage forms. The design of the novel in vitro test system
was based on the porcine oesophageal mucoadhesion test system
described by Young and Smart (1998) and Kockisch et al. (2004)
and in gastric retention studies (Riley et al., 2002). This apparatus
allows a at surface to be created by applying a gentle vacuum to
pull the oesophageal mucosa down onto the surface. Most bioadhesion measuring apparatus tends to measure tensile stresses, i.e. the
force required to pull a bioadhesive material vertically away from a
mucosa. In the oesophagus the formulation will be exposed to shear
forces relative to the mucosa, i.e. force applied horizontally across
the mucosal surface such as when food is swallowed and moved
down the oesophagus into the stomach by peristalsis; in previous

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work adhesive performance has been shown to vary between tensile and shear testing (Mortazavi and Smart, 1995). The apparatus
described will therefore use shear stresses to measure adhesive
interactions in terms of the force needed to pull a disc coated with
the test material over a attened section of oesophageal mucosa.
In this paper the development and evaluation of such a test system will be described, along with some initial studies investigating
some materials with contrasting adhesive properties.

Dispersions of HPMC (7% w/w), PEG (15% w/w), sodium alginate

(1.5% w/w), gelatin (8% w/w), FI27 (10% w/w) and PVA (4%) were
all prepared by adding the appropriate mass of polymer to rapidly
vortexing (magnetic stirrer) de-ionised water. Simulated saliva was
prepared by dissolving 0.9% sodium chloride solution in deionised
water and then adding 0.5% gastric mucin.

The surfaces of these glass discs were roughened using a low

grade, silicon carbide abrasive (Carborundum, 1000 grade) as in
preliminary studies this was found to enhance the coating procedure. This method involved grinding the surface of the glass over
the abrasive (dispersed in water), employing continuous, circular
motions. Examining the discs using an optical microscope (10)
ensured the optical uniformity of the surface roughness.
The surface roughness of the glass discs was measured using a
Talysurf (Taylor Hobson Ltd.). The roughness average (Ra ). is the
arithmetic average value of the absolute departures of the roughness prole from the mean line (the line drawn such that the sum
of the areas embraced by the line surface prole above the line is
equal to the sum of those below the line). Roughened glass discs
(O.27 Ra m1 0.013) were used as the substrate to apply coating
materials for testing in the in vitro test system. Before coating, the
glass discs were cleaned using chromic acid. All discs were soaked
in chromic acid overnight then rinsed thoroughly using (ca. 20)
copious amounts of double distilled water and oven dried at 60 C.
Films were cast from the aqueous polymer solutions using a spin
casting technique. Each glass disc was mounted horizontally into
the circular chuck of a precision spin coater (Model 6206, PI-KEM,
UK) and held in place by the application of a vacuum to the underside of the disc. A xed volume of polymer solution was placed
onto the glass disc and the disc rotated. The majority of the lms
were spun at a speed of ca. 1000 rpm for 30 s, however, the more
viscous solutions of HPMC blends and sodium alginate required a
higher rotation speed of ca. 2000 rpm for 50 s. The lms were then
air-dried (at ambient temperature) and weighed; further applications were applied until a dry constant weight of between 1.50 and
2.00 mg was achieved. Parafn wax was melted over a beaker of
boiling water. Once molten the glass discs were coated by dipping
the surface of each into the molten wax using tweezers and were
then allowed to set at room temperature. The lms were stored in
a dessicator over silica gel and used within 1 week to avoid any
discrepancies due physical ageing.

2.3. Preparation of mucosal surface

2.5. Evaluation of adhesion

Fresh porcine oeosophagi were obtained within 30 min of sacrice from P.C. Turner abattoir, Farnborough, Hants. A normal saline
solution was added to the tissues to prevent dehydration during
transit. On return to the laboratory the luminal mucosae were
individually dissected free from the outer musculature using the
following procedure. A longitudinal cut was made through both
layers of the musculature down one side of the oesophagus (using
round-ended dissection scissors), taking great care not to disrupt
the delicate epithelium. At the pharyngeal end of the oesophagus the epithelium was separated from the closely bound layers
of musculature using sharp-tipped dissection scissors. The musculature was subsequently discarded after being peeled away from
the underlying submucosa, in a single motion, using minimum
force. Tissue was then ash frozen in liquid nitrogen, and stored
in polythene bags frozen at 20 C until use.
Prior to each experiment, the epithelial tissues were thawed
by immersion in normal saline at an ambient temperature, for a
minimum period of 2 h. The epithelium was then cut open longitudinally down one side, unfolded and subsequently mounted into
the test apparatus exposing the luminal test surface. Once clamped
into position a vacuum (ca. 10 Torr) was applied to the tissue maintaining it in position throughout the testing procedure.

The apparatus consisted of the porcine oesophageal mucoadhesion test system described previously mounted on a platform
that could be lowered at a rate of 1 mL min1 constructed at the
University of Portsmouth (Fig. 1). The nylon cord passed through
a small wheel and attached to the base of a top-pan balance (GEC
Avery), linked to a PC computer to collect the data, and this was analysed using a standard spreadsheet programme (except program
in computers).
Epithelial tissues were mounted in the in vitro test celI and
allowed to equilibrate for ca. 30 min at a 10 angle (an angle the
was near to horizontal but did not allow saliva to pool on the tissue), with a ow of simulated saliva of 1 mL min1 . Initially, the
coated glass disc was attached to the cord descending from the
balance; the lid of the test cell chamber was lifted. Preliminary
studies revealed that the application of a 2 g weight to the uncoated
surface of the disc gave better more reproducible data. These circular weights were created using solder and ling to the appropriate
size, and tted to the coated glass discs just before the experiment
commenced.
The coated glass disc was placed on to the surface of the PTFE
launch using tweezers (roughened or coated surface downwards)
such that it would be pulled up the 10 incline. The lid of the test cell
chamber was replaced immediately. Shear force was then applied
by lowering the mobile platform at a rate of ca. 0.2 mm s1 and
simultaneously, the balance output was collected, recording the
force applied at intervals of 1 s for a period of 290 s. Once the cord
attached to the balance was fully taut, the PTFE launch was carefully
removed allowing the test disc and tissue to come into contact. The

2. Materials and methods

2.1. Materials
Polytetrauoroethylene (PTFE) was purchased from Goodfellow, Huntingdon. Hydroxypropylmethylcellulose (HPMC), under
the trade names Pharmacoat 615 were supplied by Harke Group,
Muelheim an der Ruhr, Germany; the Pluronic@ copolymer F127
was supplied by BASF PLC, Cheadle, Cheshire. Poly(ethylene glycol) 6000 MW grade (PEG) and Parafn wax (high melting point,
ca. 60 C) was purchased from B.D.H Chemicals Ltd., Poole.
Gelatin (type B, from bovine skin), sodium alginate (medium
viscosity grade from Mactocystis pyrifera) uorescein, Type III partially puried mucin, from porcine stomach and sodium chloride,
purchased from SigmaAldrich Chemical Company, Poole, Dorset.
Polyvinyl alcohol (PVA) 15,000 mW grade was supplied by Fluka
Chemicals, Gillingham.
2.2. Preparation of solutions

2.4. Preparation of test discs

Glass discs (8 mm diameter, 3 mm thick) were manufactured by
Sci-Iab, Bognar Regis, West Sussex, and incorporated a small metal
hook on one edge.

J.D. Smart et al. / International Journal of Pharmaceutics 447 (2013) 199203

201

Fig. 2. Force distance curve for a typical sodium alginate coated disc.

3. Results

Fig. 1. Schematic diagram of the in vitro oesophageal adhesion model.

data was then analysed to yield the maximum force required to

initiate shear motion and the total work done in moving the glass
disc a length of 59.5 mm across the epithelial surface.
It became apparent that some of the more hydrophilic materials
were being removed from the surface of the disc through a combination of hydration and friction during adhesion testing, resulting
in remnants of polymer being deposited on the tissue for prolonged
periods of time. The presence of such polymer residue could affect
the adhesion of other polymers tested on the same tissue. It was
therefore decided that only one material would be tested per tissue.
To take into account tissue variances an identical uncoated roughened glass disc was tested on each tissue before adhesion testing
with the coated test disc.
Each material was tested six times on six different tissues and
shows the average the work done and maximum detachment force
values calculated over 290 s for all of the materials assessed. The
corresponding values of the roughened glass controls are also
included for each material.

The force distance curve for two coatings, one a putative

adhesive and the other a non-adhesive material are shown in
Figs. 2 and 3, along with the roughened glass disc control. In
the case of the known mucosadhesive sodium alginate, force is
required initially to break the adhesive bond, and once broken the
bonds reformed and then broke again (Fig. 2). In contrast, the nonadhesive PEG showed adhesive properties similar to that of the
glass disc (Fig. 3).
From these traces the maximum detachment force was found as
the greatest resistance force to movement across the tissue, while
the work done was the area under the force distance curve for a
distance of 59.5 mm. The work done and maximum detachment
force values for the roughened glass controls appear to vary to
some extent between the tissues used for the individual materials (one-way ANOVA, p < 0.05). These differences are attributed to
tissue irregularities, anticipated when working with biological tissues from different animals. In order to minimise these differences
related to tissue topography the test values were normalised by
dividing by the values obtained for the uncoated glass discs when
initially applied to the same tissue to give the relative adhesive
performance.
A range of coating materials with various properties were tested
in this system (Table 1), and their relative adhesive performance
determined (Fig. 4). The greatest resistance to movement was
shown by the sodium alginate coated discs, and the least by those
coated with parafn wax (Multiple comparison Tukeys HSD,
p < 0.05).
When uorescein was incorporated into the coat, a long uorescent trail was detected on the tissue surface after testing (433 mm
long for F127 and 263 mm long PEG). There was evidence that some

2.6. Polymer retention on the test disc

Further adhesion experiments were carried out to investigate
the extent of polymer loss from the coated glass substrate during
testing. Two polymers were selected for this investigation, F127
and PEG. Solutions of these polymers were prepared in a solution
of 103 M sodium uorescein in double distilled water and lms
were cast onto the glass substrate.
Adhesion experiments were carried out with the polymer/uorescein coated discs. At the end of each experiment the
humidier and articial saliva ows were terminated, the tissue
removed and spread out on a at clean surface. An ultra-violet lamp
was then used to detect traces of uorescein present on the tissue
surface. Observations recorded included; the length of uorescein
deposition along the tissue surface, presence of uorescein on the
glass substrate and percentage polymer mass loss from the glass
disc.

Fig. 3. Force distance curve for a typical PEG coated disc.

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J.D. Smart et al. / International Journal of Pharmaceutics 447 (2013) 199203

Table 1
Average work done (WD) and maximum detachment force (MDF) values for all the materials and rouqhened qlass controls evaluated in the in vitro test system (n = 6).
Material

Film weight (mg) (s.d.)

WD (J)

s.d.

MDF (mN)

s.d.

Sodium alginate
Glass control
PVA
Glass control
HPMC
Glass control
F127
Glass control
Gelatin
Glass control
PEG
Glass control
Parafn wax
Glass control

1.96 (0.09)

4154.03
666.35
625.23
628.94
1209.28
665.01
1121.57
526.20
575.25
605.51
591.50
585.49
416.44
532.32

1379.01
72.71
57.00
73.54
339.70
118.83
361.05
59.86
76.02
70.79
98.85
54.69
54.84
57.52

263.42
13.72
29.11
13.44
95.14
14.52
81.85
11.56
19.83
12.58
12.70
12.86
8.61
10.60

111.39
1 .11
8.36
2.99
43.68
2.52
37.57
1.92
7.61
2.03
1.84
2.05
1.35
1.42

1.92 (0.10)
1.73 (0.18)
1.82 (0.17)
1.89 (0.20)
1.83 (0.16)
3.5 (0.20)

dye was retained on the disc with F127, whereas none was evident for PEG, indicating that all the coating had been removed.
This observation was conrmed by drying the discs and reweighing, demonstrating that 97% of the weight of the coat was removed
for PEG, whereas for F127 82.7% (SD 24.16) was removed.
4. Discussion
The aim of this study was to develop a test system to evaluate
adhesion of solid oral dosage form coatings to oesophageal mucosa
using shear stresses. Two materials were investigated in the initial
experiments; these were the high molecular weight sodium alginate, with established bioadhesive properties (Smart, 2005), and
PEG, a relatively low molecular weight non-adhesive material. In
a typical graph, there would be an initial increase from zero in the
force measured as the tension is applied to the cord and joint. If
no adhesion occurs then the force measured will correspond to
the frictional force of pulling the disc across the mucosa as seen
in the controls. If adhesion occurs then there would be a signicant
increase in the force measured relative to the control. In the case
of sodium alginate, such adhesion of the lm-coated disc to the
mucosal surface was evident, and once broken the adhesive joint
reformed when moved down the tissue until rebroken (Fig. 2). This
is the pattern one might expect of an adhesive material, if dislodged
it will simply re-adhere at the next contact point. PEG was expected
to hydrate rapidly and form a lubricating lm between disc and
mucosa; but the graph obtained with this showed little change to
that of the disc alone (Fig. 3). This coating is being rapidly removed
in an aqueous environment and therefore the forces generate are
similar to the glass disc alone.

Fig. 4. Relative work done and maximum detachment force for a range of test
coatings (n = 6, SD bars).

A variety of materials were tested in this system that should

have both adhesive or lubricant properties (Table 1, Fig. 4). Sodium
alginate is a polysaccharide consisting of homopolymeric blocks of
mannuronate and guluronate residues, and is known to be strongly
a bioadhesive in tensile testing studies (Mortazavi and Smart, 1995;
Smart, 2005). Sodium alginate lms were seen to facilitate adhesion, and this property might be of use if it is planned to retain
a formulation within the oesphagus. HPMC is a cellulose derivative that has been used extensively in pharmaceutical products
and is employed as the bioadhesive in some buccal formulations
(e.g. Suscard BuccalTM ), and has been found to be bioadhesive in
tensile testing studies (Mortazavi and Smart, 1995). This material
also showed signicant adhesive properties, indicating that tablets
coated with this material may lodge in the oesophagus. F127 is a
pluronic surfactant consisting of two blocks of hydrophilic polyoxethylene anking a central hydrophobic polyoxypropylene chain,
and it was anticipated that the hydrophilic components may orientate themselves so that they were projected outwards from the
lm to form a hydrated lubricating coating. However, this material
was found to act as a wet adhesive like other hydrophilic macromolecules and adhere to the oesophagel mucosa. Further work will
look at different grades of pluronics with differing HLB values to
see if they behave differently. Gelatin is protein that is the main
component of soft and hard capsules and has been reported to
have some bioadhesive properties, but in this study little adhesion
was observed. Low molecular weight PEG and PVA are hydrophilic
non-ionic polymers that have been reported to be non adhesive,
and indeed hydrate rapidly to form a lubricating lm. These lms
were found to behave in a similar fashion to the uncoated disc in
this study. The dye studies suggest that such polymers are rapidly
removed from the surface of the glass disc and deposit onto the
tissue in this study. Parafn wax was used to model the waxes
used in many formulations (Drake et al., 2002). As it is hydrophobic
and would provide a smooth surface it would be predicted not to
adhere, and the maximum detachment force and work done was
reduced relative to the glass disc alone, although the effect was not
signicant (multiple comparison Tukeys HSD, p < 0.05).
From this work it was concluded that this test system was capable of discriminating between polymer coatings, and showed that
some had signicant adhesive properties in the oesophagus. However none were identied as having signicant lubricating effects
relative to a roughened glass disc control. In further work a range
of materials will be evaluated for their lubricating and adhesive
effects, as a means of enhancing retention or facilitating transport of
a solid dosage form through the oesophagus. Other factors affecting
adhesion, such as the ow rate of liquid over the surface, pretreatment of the oesophagus and the thickness of the lm coating, will
also be evaluated.

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References
Al-Dujaili, H., Florence, A.T., Salole, E.G., 1986a. In vitro assessment of the adhesiveness of lm-coated tablets. Int. J. Pharm. 34, 6774.
Al-Dujaili, H., Florence, A.T., Salole, E.G., 1986b. The adhesiveness of proprietary
tablets and capsules to porcine oesophageal tissue. Int. J. Pharm. 34, 7579.
Batchelor, H.K., Banning, D., Dettmar, P.W., Hampson, F.C., Jolliffe, I.G., Craig, D.Q.M.,
2002. An in-vitro mucosal model for the prediction of the bioadhesion of alginate
solutions to the oesophagus. Int. J. Pharm. 238, 123132.
Channer, K.S., Virjee, J.P., 1985. The effect of formulation of oesophageal transit. J.
Pharm. Pharmacol. 37, 126129.
Collaud, S., Warloe, T., Jordan, O., Gurny, R., Lange, N., 2007. Clinical evaluation
of bioadhesive hydrogels for the topical delivery of hexylaminolevulinate to
Barretts esophagus. J. Contr. Rel. 123, 203210.
Drake, W.M., Worsley, D.F., Lentle, B.C., Kendler, D.L., 2002. Monitoring Esophogeal
trasit of wax-polished alendronate in healthy post-menopausal women: a new
technique to study of pill transit time. Curr. Ther. Res. 63, 103109.
Hill, L.L., Woodruff, L.H., Foote, J.C., Barreto-Alcoba, M.A., 2005. Esophageal injury by
apple cider vinegar tablets and subsequent evaluation of products. J. Am. Diet.
Assoc. 105, 11411143.
Kockisch, S., Rees, G.D., Young, S.A., Tsibouklis, J., Smart, J.D., 2004. In-situ evaluation of drug-loaded microspheres on a mucosal surface under dynamic test
conditions. Int. J. Pharm. 276, 5158.
McCargar, L., Crail, D., Dansereau, R., Myers, M., Lane, M., 2001. The in-vitro porcine
model is not predictive of the esophageal transit of risedronate tablets in
humans. Int. J. Pharm. 222, 191197.
Mako, A., Csoka, G., Pasztor, E., Marton, S., Horvai, G., Klebovich, I., 2009. Formulation
of thermoresponsive and bioadhesive gel for the treatment of oesophageal pain
and inammation. Eur. J. Pharm. Biopharm. 72, 260265.

203

Mortazavi, S.A., Smart, J.D., 1995. An assessment of some factors inuencing the
in-vitro assessment of mucoadhesion. Int. J. Pharm. 116, 223230.
Jaspersen, D., 2000. Drug induced oesophageal disorders. Drug Saf. 22 (3), 237
249.
Marvola, M., Rajaniemi, M., Marttila, E., Vahervuo, K., Sothmann, A., 1982. Effect of
dosage form and formulation factors on the adherence of drugs to the oesophagus. J. Pharm. Sci. 71 (9), 975977.
Perkins, A.C., Frier, M., Blackshaw, E., Spiller, R.C., Fairbairn, K.J., Dansereau, R.J.,
Kinghorn, T., San, P., Hosking, D., 2006. Esophageal transit of the weekly lm
coated risedronate (Actonel )placebl tablet in subjects with Kyphosis. Int. J.
Pharm. 311, 2025.
Perkins, A.C., Wilson, C.G., Frier, M., Blackshaw, P.E., Dansereau, R.J., Vincent, R.M.,
Wenderoth, D., Hathaway, S., Li, Z., SpillerF R.C., 2001. The use of scintigraphy to
demonstrate the rapid esophageal transit of the oval lm coated placebo risedronate tablet compared to a round uncoated placebo tablet when administered
with minimal volumes of water. Int. J. Pharm. 222, 295303.
Riley, R.G., Smart, J.D., Tsibouklis, J., Young, S.A., Hampson, F., Davis, J.A., Kelly, G.,
Dettmar, P.W., Wilber, W.R., 2002. The gastric mucosal retention of 14 C-labelled
poly(acrylic acids): an in vitro study. Int. J. Pharm. 23 (6), 8796.
Sakkinen, M., Marvola, J., Kanerva, H., Lindevall, K., Ahonen, A., Marvola, M., 2004.
Scintigraphic verication of adnerence of chitosan formulation to the human
oesophagus. Eur. J. Pharm. Biopharm. 57, 145147.
Smart, J.D., 2005. The basics and underlying mechanisms of mucoadhesion. Adv.
Drug Deliv. Res. 57, 15561568.
Ueda, K., Muto, M., Chiba, T., 2011. A case of esophageal ulcer caused by alendronate
sodium tablets. Gastrointest. Endosc. 73, 1037.
Young, S.A., Smart, J.D., 1998. The porcine oesophageal mucoadhesion test system:
a novel in vitro apparatus for the evaluation of liquid and semisolid retention. J.
Pharm. Pharmacol. 52, s167.