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Journal of the Ghana Science Association, Vol. 4 No. 1. 2002

Responses of saliva pH to ingestion of Golden Tree

chocolate and the effect of stick chewing
F.K. Addai1, I.K. Nuamah2 and Joyce Addo3
1

Department of Anatomy, University of Ghana Medical School,

P. O. Box 4236, Accra Ghana
2
15 St. Chad Road, Withington, Manchester, M20 4WH, England
3
111 Lake Park Drive, Martin, TN 38237, U.S.A
ABSTRACT
Volunteers ingested Golden Tree chocolate with the expectation of causing saliva pH decline, and to
determine whether stick chewing could reverse it. Saliva pH rose from a median baseline of 7.30 (mean
7.32, SD 0.34) to 7.80 (mean 7.82, SD 0.34) in volunteers (stick chewers) who chewed a piece of
Garcinia manii stick for 10 minutes immediately after chocolate ingestion. In volunteers (controls) who
did not chew stick after chocolate ingestion, saliva pH rose from a median baseline of 7.50 (mean 7.44,
SD 0.32) to 7.60 (mean 7.61, SD 0.25). In all volunteers, saliva pH fell from peak values over a 20minute period. However, whereas the median saliva pH of control fell significantly (P<0.01) below
baseline value, that of stick chewers did not; although salivary pH in both groups remained higher than
neutral throughout the experiment. Each of thirty-eight voluntees ingested a bar of Golden Tree
chocolate (pre-warmed in an oven to prevent chewing) after giving a baseline saliva sample. The pH of
saliva samples was measured using a Kent EIL 7020 pH meter. Our results suggest that Golden Tree
chocolate has a non-acidogenic potential that was transiently accentuated by stick chewing.
INTRODUCTION
Golden Tree milk chocolate produced
by Cocoa Processing Company in Ghana has a
sweet taste, and the ingredients listed on its
wrapper include sugar. For these reasons, it was
selected for ingestion with the intention of
depressing saliva pH and subsequently to assess
the efficacy of stick chewing in reversing the pH
decline. The rationale was that sugar in Golden
Tree chocolate would be substrate for oral
bacteria to break down and release organic
(lactic, Propionic, and acetic) acids in
consonance with the acidogenic basis of dental
caries formation (Pollard et al., 1996).
Previously, it had been shown that stick
chewing reversed salivary pH decline following
a glucose challenge (Addai et al., 2002). The
present study was a follow-up to determine
whether or not a similar effect could be
demonstrated after ingestion of a sucrosesweetened snack. We have defended the choice
of the pH of saliva in our experimental design
(Addai and Nuamah, 2000).
Briefly, the
response of salivary pH is important because

saliva is the source of components that affect

plaque pH (Mandel, 1994). Plaque is the
mixture of food remnants and oral
microorganisms that adhere to teeth, the
increasing acidity of which initiates the carious
process culminating in tooth decay (Anderson et
al., 1993). Saliva pH can also decrease the acid
challenge of cariogenic bacteria by buffering
their acids to a pH that will not demineralise
teeth (Anderson et al., 1993).
Chocolate contains more than three
hundred known chemicals (Spadaccini, 1998),
some of which are implicated in its hedonic or
pleasurable effects (Bruinsma and Taren, 1999).
A number of bad outcomes have been linked
with eating chocolate; including acne. tooth
decay, and weight gain. However, chocolate is
also known for being an anti-depressant, an
aphrodisiac, and capable of lowering the risk of
heart disease (Spadaccini, 1998). Available
literature (in English) on the causal relationship
between chocolate ingestion and formation of
dental caries appears to be inconsistent. In one

Journal of the Ghana Science Association, Vol. 4 No. 1. 2002

study of ten snack foods, milk chocolate caused

the greatest pH drop in in vivo dental plaque
after ingestion (Koparal, et al., 1998). In
another investigation of six foods, chocolate bar
produced the second highest quantity of lactic
acid thirty minutes after ingestion, but the third
lowest quantity of acid two hours after ingestion
(Linke et al., 1997). It has also been pointed out
that the sugar in chocolate does not contribute to
cavity formation any more than the sugar in any
other food (Spadaccini, 1998). Moreover, some
reports suggest that the view that sucrose is a
veritable cariogenic agent may not be entirely
accurate. For instance, clearance of sucrose
from saliva proceeded much faster than glucose,
fructose, maltose or sorbitol (Luke et al., 1999);
whereas oral clearance of foods containing
cooked starch was significantly slower and
contributed to long periods of lactic acid
production compared to chocolate-covered
candy bar (Linke and Birkenfeld, 1999); and
high-starch foods contributed more significantly
to the development of carious lesions than highsucrose foods (Kashket, et al., 1996).
Golden Tree milk chocolate bears a label
indicating that it contains a minimum of 35%
cocoa solids. Undoubtedly, the variety of
ingredients in chocolate preparations around the
world makes it difficult to determine which
component affects oral pH. In any case, there
appears to be no study of the direct effect of
Golden Tree chocolate ingestion on plaque or
salivary pH so as to assess its harmfulness or
otherwise to teeth. This study therefore fills an
information gap that is unhelpful to the
promotion of dental health in Ghana. Besides,
the present report is important to the health
educator who needs to give dietary advice. It is
pertinent in this context, that dental caries
incidence among Ghanaians is high (Addo,
1997) and possible dietary factors responsible
for this state of affairs need to be identified.

volunteers before the experiment were the same

as previously described (Addai and Nuamah,
2000). The sample size was determined to have
a 90% power of detecting a difference of 0.40 in
the mean pH of saliva samples obtained from
the volunteers at 1% significance level and a
standard deviation of 0.30. The experiments
were done between 12 noon and 2pm. At the
commencement of the experiment, each
volunteer gave a saliva sample (T0) by spitting
3-4 times into a labelled 25 ml clean and dry
conical flask. After giving the T0 saliva sample,
each subject was given a 100 g bar of Golden
Tree Chocolate to eat within 15 minutes. To
prevent chewing, the chocolate was pre-warmed
o
(32 C) in an oven until it was soft to touch.
Immediately after ingesting the chocolate, stickchewing volunteers were given a piece of
chewing stick (Garcinia manii, local name
Nsokor) to chew for 10 minutes. Control
volunteers did not chew anything; instead they
waited to give saliva samples (T1)
synchronously with stick-chewers at the end of
10 minutes. Thereafter, every volunteer gave a
saliva sample at five-minute intervals for a
further 20 minutes. The Golden Tree chocolate
was purchased from a Mobilmart supermarket,
and a chewing stick billet bought at the
Kantamanto market in Accra after a botanist had
identified the species.
Each appropriately
labelled saliva sample was promptly diluted
with 20 ml distilled water (pH 7.0) to facilitate
pH measurement using a Kent EIL, 7020 pH
meter. The pH of saliva samples collected from
each
volunteer
was
separately
and
independently measured by FKA who was
blinded to the group they belonged to. The pH
values from all subjects were analysed on an
IBM personal computer for differences in their
medians/means, using Instat-2 statistical
software.
3.

2.

MATERIALS AND METHODS

Thirty-eight volunteers from the
population of students at the University of
Ghana Medical School were randomly divided
into stick chewers and controls by balloting.
Inclusion criteria and details of preparing the

RESULTS
The time interval for collecting saliva
samples and their designation to permit
distinction are given in Table 1. Median and
mean pH of saliva samples obtained at various
time
intervals
in
this
study
are

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Journal of the Ghana Science Association, Vol. 4 No. 1. 2002

Table 1. Time intervals for collecting saliva samples from volunteers. The figures in brackets
indicate the cumulative time after chocolate ingesting by control subjects who did not chew stick.
Activity

Designation of saliva sample

Beginning (baseline)
Ingestion of Golden Tree Chocolate
Chocolate effect I/Stick chewing
Chocolate effect II/Stick effect I
Chocolate effect III/Stick effect II
Chocolate effect IV/Stick effect III
Chocolate effect V/Stick effect IV

Elapsed time In
minutes
----15
10
5(15)
5 (20)
5(25)
5(30)

T0
---T1
T2
T3
T4
T5

Table 2. Basic statistics on pH of whole saliva collected from volunteers and designated as
indicated in Table 1.
Status of volunteers

*N

Statistic

T0

T1

T2

T3

T4

T5

Stick chewers

19

Median
Mean
S.D

7.30
7.32
0.34

7.80
7.82
0.34

7.60
7.58
0.31

7.50
7.44
0.26

7.40
7.40
0.30

7.30
7.35
0.26

Median
7.50 7.60 7.50
Mean
7.44 7.61 7.48
S.D
0.32 0.25 0.18
*N represents sample size, and S.D. stands for standard deviation.

7.40
7.43
0.19

7.40
7.41
0.16

7.30
7.35
0.17

Controls

19

presented in Table 2.
A comparison by
Friedmans Nonparametric Repeated Measures
test yielded a statistic of 53.80 for the pH of
saliva collected at various time intervals from
stick chewers and 25.76 for saliva from
controls. These Friedman statistics (for both
groups of volunteers) had probability or P<
0.0001 indicating that changes in salivary pH
from baseline values were statistically
significant. The pH of saliva collected from
stick chewers and controls at each time point of
the experiment were compared using unpaired
Welchs approximate t-test (that does not
assume equal variances) and summarized in
Table 3. Only the pH of T1 saliva samples for
stick-chewers
was
significantly
higher
compared to that of control volunteers (Table 3).
Thirty minutes after ingesting chocolate the
salivary pH of stick chewers (T5) was
significantly higher than their baseline sample
(T0) becauses Dunns multiple comparison

posthoc test produced a rank sum difference of

44.0 (P<0.01). For controls, the pH of saliva
thirty minutes after chocolate ingestion (T5) was
lower than their baseline (T0) sample, with a
Dunns multiple comparison posthoc rank sum
velue of negative 76.50 and P < 0.001.

4.

DISCUSSION
A non-parametric analysis was used to
compare the median pH of saliva collected at
the various time-points because a Bartletts test
for homogeneity of variances indicated
significant differences among their standard
deviations,
making
ANOVA
method
inappropriate. The finding that pH of saliva was
raised significantly above that of basal level
after control volunteers ingested Golden Tree
chocolate was against our original expectation.
But it demonstrates that ingestion of Golden
Tree chocolate

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Journal of the Ghana Science Association, Vol. 4 No. 1. 2002

Table 3.
Synopsis of comparisons by the Welchs unpaired approximate t-test of mean pH of
saliva collected from stick chewers and controls.
Saliva Samples
Statistic

T0

T1

T2

T3

T4

T5

Mean pH difference
Degrees of freedom
t-value

0.12
35
1.09

-0.21
32
2.17

-0.11
29
1.27

-0.005
33
0.07

0.02
27
0.21

-0.005
31
0.07

Controls

-0.10
-0.41
-0.27
-0.16
-0.14 -0.15
0.33
-0.13
0.06
0.15
0.17 0.14
Probability
0.28
0.04
0.21
0.94
0.84 0.94
Mean pH difference is the Mean saliva pH of controls minus Mean saliva pH of stick chewers.
failed to cause a decline in saliva pH, instead it
raised it. Thus, Golden Tree chocolate is not
only non-acidogenic it is apparently antiacidogenic. This finding is interpreted to mean
that ingesting Golden Tree chocolate is not
harmful to teeth, but may actually protect
against dietary acidogenicity.
It is inferential from our results that
Golden Tree chocolate contains some
ingredient(s) that either prevented its sugar from
being digested by oral bacteria or neutralised the
acid resulting from sucrose metabolism.
Regardless of the mechanism responsible for the
present finding, it is hypothesised that the high
content of cocoa solids in Golden Tree
chocolate could be a contributory factor to its
demonstrated non-acidogenicity. It is of interest
that cocoa has cariostatic properties attributable
to antibacterial effects of cocoa mass extract
(Takahashi et al., 1999) and cocoa bean husk
extract shows caries inhibitory activity
(Ooshima et al., 2000a). Indeed cocoa mass
extract has also been reported to have direct
cariostatic activity (Ooshima et al., 2000b).
Moreover, there are indications that the cocoa
butter in chocolate coats the teeth and may help
protect them by preventing plaque from forming
(Spadaccini, 1998).
The experience of enhanced salivation
following ingestion of Golden tree chocolate is
commonplace. This may be partly ascribed to
the habit of chewing the chocolate, since
mastication results in increased salivation

(Edgar, 1992). In the present study mastication

was eliminated, so increased salivation would
have resulted from gustatory (taste) and
olfactory (smell) stimulation. Again, it is
common knowledge that Golden Tree chocolate
has a strong cocoa aroma, and raw cocoa beans
have a sour taste. It is noteworthy therefore that
taste stimuli can cause a ten-fold increase in
saliva flow, and a sour stimulus is the most
effective in accelerating saliva flow (Edgar,
1992). The benefit of increased salivary flow
on caries inhibition is well known (Humphrey
and Williamson, 2001).
The rise in saliva pH was maintained
significantly higher in stick chewers compared
to those of controls. However, it is noteworthy
that both groups of volunteers could not be at
risk of caries formation since the pH of their
saliva was still in a safe range (>7.0) thirty
minutes after eating chocolate. considering that
only T1 saliva samples obtained immediately
after stick chewing had a statistically higher pH
than that of controls, it is suggested that the
accentuation of saliva pH elevation resulting
from stick chewing was transient. Inferentially,
the return of saliva pH to basal level was
apparently faster in stick chewers compared to
controls. Why this happened is difficult to
explain. It is however, attractive to speculate
that swallowing occurred faster than salivation
after chewing ceased, or that relatively higher
pH levels (mean 7.82, SD 0.34) obtained from
stick-chewers engendered a faster decline to

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Journal of the Ghana Science Association, Vol. 4 No. 1. 2002

basal level. Nonetheless, it is interesting that

the pH of the last saliva sample (T5) was
statistically lower than baseline (T0) in controls
but higher than T0 in stick chewers. There is a
suggestion in this result that chewing stick after
Golden Tree chocolate ingestion may be
superfluous as it does not offer any significant
or long-lasting additional dental protection
against acidogenic agents. However, it may be
useful to study whether chocolate ingestion
alone can reverse a decline in salivary pH
following a glucose rinse, a was demonstrated
with stick chewing (Addai et al., 2002).
5.

CONCLUSION
It is concluded that ingestion of Golden
Tree chocolate did not cause anticipated decline,
instead it raised the pH of saliva suggesting that
it is not only non-acidogenic but may have antiacidogenic potential. Considering that only a
transient accentuation was obtained from stick
chewing in raising salivary pH after ingestion of
Golden Tree chocolate, chewing stick after
chocolate ingestion may be superfluous. The
ingestion of Golden Tree chocolate may be
recommended as an adjunct to other cariesprevention oral hygiene methods, and
particularly as the snack of choice to reduce
cariogenic challenge to teeth after meals.
ACKNOWLEDGEMENTS
The financial support of the Dreyfus
Health Foundation of New York is
acknowledged. We thank the volunteers from
the year classes of 1996, 1997, and 1998 of the
University of Ghana Medical School who
participated in this study. We are also grateful
to our two Research Assistants, Ms. Victoria
Andan and Mr. Alex Prince Egyir Yawson. Dr.
G.K. Ameka of the Department of Botany,
University of Ghana, Legon, identified the
species of chewing stick.

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