Archives of Oral Biology (2005) 50, 541552

www.intl.elsevierhealth.com/journals/arob

Erosive effects of different acids on bovine enamel:

release of calcium and phosphate in vitro
Christian Hannig*, Arne Hamkens, Klaus Becker,
Rengin Attin, Thomas Attin
Department of Operative Dentistry, Preventive Dentistry and Periodontology,
ttingen, Robert-Koch-Str. 40, D-37075 Go
ttingen, Germany
University of Go
Accepted 2 November 2004

KEYWORDS
Erosion;
Calcium;
Phosphate;
Demineralisation

Summary The present study intended to investigate minimal erosive effects of

different acids on enamel during short time incubation via determination of calcium
and phosphate dissolution. Bovine enamel specimens were eroded for 15 min with
eight different acids of pH 2, 2.3 and 3 (citric (CA), maleic (MA), lactic (LA), tartaric
(TA), phosphoric (PA), oxalic (OA), acetic (AA) and hydrochloric acid (HCl)). Calcium
(Ca) and phosphate (P) release were determined photometrically using arsenazo III
(calcium) and malachite green (phosphate) as substrates. Each subgroup contained
eight enamel specimens. Amount of titratable acid was determined for all acidic
solutions.
MA, LA, TA, AA and HCl caused linear release of Ca and P, PA of Ca, CA of P. For CA,
MA, LA, TA, AA, PA and HCl mineral loss was shown to be pH-dependent. Ca dissolution
varied between 28.6
4.4 (LA, pH 2) and 2.4
0.7 nmol mm2 min1 (HCl, pH 3), P
dissolution ranged between 17.2
2.6 (LA, pH 2) and 1.4
0.4 nmol mm2 min1
(HCl, pH 3). LA was one of the most erosive acids. AA was very erosive at pH 3. HCl and
MA were shown to have the lowest erosive effects. There was only a weak correlation
(r = 0.28) between P and Ca release and the amount of titratable acid.
The method of the present study allows investigation of minimal erosive effects via
direct determination of P and Ca dissolution. During short time exposition at constant
pH level, erosive effects mainly depend on pH and type of acid but not on amount of
titratable acid.
# 2004 Elsevier Ltd. All rights reserved.

Introduction
* Corresponding author. Tel.: +49 551 39 2898;
fax: +49 551 39 2037.
E-mail address: Christian.hannig@med.uni-goettingen.de
(C. Hannig).

Dental erosion is defined as irreversible loss of

dental hard tissue due to chemical processes without involvement of microorganisms.1 Acids are
assumed to be the main etiological factor.2 Erosive

00039969/$ see front matter # 2004 Elsevier Ltd. All rights reserved.
doi:10.1016/j.archoralbio.2004.11.002

542

tooth destruction is classified to be intrinsic or

extrinsic according to the source of the acids.3
Main intrinsic factor inducing dental erosions is
gastric juice which comes in contact with the dentition especially in patients suffering from anorexia
nervosa, bulimia or chronic regurgitations.4,5 Erosive potential of gastric juice is mainly determined
by hydrochloric acid: gastric juice has been shown to
have a significantly higher erosive potential compared to carbonated acidic drinks.6
An external factor creating dental erosions is
represented by acidic fumes in chemical industry,
for example, in production of batteries.7,8 Also,
some orally applied medicaments, such as acetylic
salicylic acid, may cause erosions.8 However, in
modern western societies diet can be regarded as
most important factor for erosions.3,9,10 Increasing
consumption of acidic beverages, such as sport
drinks or lemonades, is one main reason.11 Also
fruits, vegetables and especially juices which are
regarded as healthy food, may cause significant
erosive effects.2,3 Thereby, citric acid represents
the main acid in fruits and vegetables.10 Vinegar and
vinegar conserves of vegetables contain acetic acid.
Oxalic acid occurs mainly in rhubarb, tartaric acid in
wine and grapes, maleic acid in apples and apple
drinks.12,13 Phosphoric acid is used in numerous cola
drinks.13 Lactic acid which is usually associated with
caries is present in sauerkraut or yoghurt as metabolic product of lactobacilli. Organic acids, in particular citric, maleic and tartatic acid, are deemed
to be extremely erosive to dental hard tissues due to
the ability of some of them to form chelate complexes with calcium released from the tooth.14
Many in vitro investigations were performed to
elucidate the erosive capabilities of different foods
or beverages which represent usually a mixture of
different acids. Typical methodical approaches are
electron microscopy, profilometric evaluation of
surface loss or determination of microhardness.15
20
Other studies already investigated erosive potential of acidic solutions measuring calcium and phosphate dissolution.21,22 This was done with various
drinks but not with pure solutions of certain acids. In
these studies, there was no clear relationship
between acidityeither described by pH or the
titratable amount of acidand demineralization.
This may be explained by other minerals or molecules present in the drinks altering the erosive
potential. As yet, the erosive potential of pure
solutions of different acids at constant pH values
typically occurring in acidic drinks has not been
characterized systematically. This is especially true
for short time incubation. Typical incubation times
in the existing studies range between 10 and
120 min, but usually incubation for 30 or 120 min

C. Hannig et al.
is quite common.13,1520 These time periods are
much longer than the contact of the teeth with
an erosive substrate during consumption of erosive
food or beverages.14,23 Furthermore, erosive effects
were often evaluated without any standardization
of pH of the test solutions.13,1520
Therefore, the present study aimed to quantify
demineralization of enamel samples via highly sensitive determination of calcium and phosphate dissolution caused by certain acids after short time
erosion for 15 min. This was done systematically
for different solutions of pure acids typically occurring in foods and drinks at constant pH levels to
provide standardized basic parameters for direct
comparison of the erosive effects.

Materials and methods

Specimens
Cylindrical enamel specimens (diameter 5 mm)
were gained from bovine incisors of 2-year-old cattle. Specimens were etched laterally and at the
bottom for 30 s with 37% phosphoric acid gel (Etching gel, DMG, Hamburg, Germany) and treated for
30 s with primer (OptiBond FL, bottle 1, Kerr, Karlsruhe, Germany). Bonding of OptiBond (bottle 2) was
applied afterwards and light cured for 30 s. Application of bonding was performed three times. In the
following, unsealed enamel surfaces were ground
flat and polished (grit 4000). In a standardized
grinding procedure, ca. 200 mm of enamel was
removed. This was controlled with a micrometer
measurement device (HHW, Hommel, Schwenningen, Germany). Specimens with structural alterations of enamel were eliminated.
Before incubation with the different acids, specimens were stored in artificial saliva for 7 days for
standardized remineralization.24 Five hundred millilitres of artificial saliva contained 0.001 g ascorbate,
0.015 g glucose, 0.290 g NaCl, 0.085 g CaCl2, 0.080 g
NH4Cl, 0.635 g KCl, 0.080 g NaSCn, 0.165 g KH2PO4,
0.100 g carbamide and 0.170 g Na2PO4.
Eight enamel specimens were included in each
subgroup.

Acids
Enamel specimens were incubated in the acidic solutions of acetic, citric, maleic, tartaric, phosphoric,
oxalic, hydrochloric and lactic acid (Merck, Darmstadt, Germany) at pH 2, 2.3 and 3, respectively.
Concentrations of the certain acids at the three pH
levels are given in Table 1. pH was checked with a pH
meter (RadiometerA/S, Copenhagen, Denmark).

Erosive effects of different acids on bovine enamel

Table 1 Concentrations of the acids at the differnt pH

levels

543

Table 2 Amount of titratable acid as determined for

the tested acids [mval NaOH/l acid]

Concentrations of acids
[mmol/l] at
Hydrochloric acid
Acetic acid
Lactic acid
Maleic acid
Tartaric acid
Phosphoric acid
Citric acid
Oxalic acid

Amount of titratable acid

[mval NaOH/l acid]

pH 2.0

pH 2.3

pH 3,0

10.5
2214
292
15.7
50.0
16.2
95.2
8.84

4.50
721
71.5
5.67
18.4
5.90
24.2
4.46

0.75
34.9
3.60
0.86
1.32
0.88
1.87
0.75

Hydrochloric acid
Acetic acid
Lactic acid
Maleic acid
Tartaric acid
Phosphoric acid
Citric acid
Oxalic acid

pH 2.0

pH 2.3

pH 3.0

10.0
2210.0
242.5
30.0
99.3
27.0
275.0
18.8

4.7
712.5
57.5
11.0
36.3
9.5
67.5
9.3

1.0
36.0
2.8
1.7
2.8
1.4
5.3
1.6

Titration performed until pH 7.

Incubation of specimens with the different

acids
Enamel samples were incubated for 5 min in 1000 ml
acid (pH 2 and 2.3) and in 5000 ml (pH 3), respectively. Due to the lower acid concentration at pH 3,
the higher incubation volume was chosen at this pH
level to avoid consumption of acid. Stability of pH
under the chosen conditions was verified in preliminary trials. During incubation and before determination of calcium and phosphate, samples were
shaken gently.

Photometric determination of calcium and

phosphate release
Mineral dissolution caused by the different acids was
determined by assessing calcium and phosphate
release into the solutions photometrically in double
assays.
Calcium (Ca) assay
Release of calcium was measured for specimens
incubated in acetic, maleic, tartaric, phosphoric,
hydrochloric and lactic acid using the arsenazo III
method (Fluitest 1, Ca-A-II, analyticon, Lichtenfels, Germany).
Arsenazo III reacts with calcium in an acid solution to form a blue purple complex. Intensity developed is proportional to the calcium concentration.
Absorption can be determined at l = 650 nm.25,26
Reagent for determination of calcium was composed
of 100 mmol/l Imidazol buffer (pH 6.5) and
0.12 mmol/l arsenazo III. Determination of calcium,
dissolved by oxalic acid and citric acid, was not
possible, since the method did not allow reliable
calcium determination in these acids.
Phosphate (P) assay
Erosive dissolution of phosphate was tested with all
acids except for phosphoric acid as this acid releases

phosphate by itself and thereby interferes with the

determination of phosphate release from enamel.
Malachite green reacts with phosphate to a coloured
complex which can be determined photometrically
at l = 650 nm. 0.045 mg of malachite green soluted
in 100 ml aqua bidest. were admixed to 12.69 g
ammonium molybdate dissolved in 300 ml HCl
(4 mol/l). The reagent was stirred for 30 min afterwards and filtered (pore size 0.22 mm). The reagents
were stored at 4 8C.
For determination of Ca and P loss, individual
standard curves were obtained for each pH of all
acids with standardized Ca and P solutions. Diluted
samples of standardized calcium and phosphate
solutions were admixed to the certain acidic solutions. Standard solutions contained 1.46 g P or Ca
per 100 ml aqua bidest. and the corresponding test
volume of acid, respectively. Precision of the measurements was validated with the standard solutions.
Phosphate release induced by phosphoric acid
and calcium dissolution caused by citric acid were
calculated according to the calcium to phosphate
ratio which was found for all other acids. Oxalic acid
was not considered for detailed determination of
phosphate release per minute due to the special
kinetics of demineralization. Mineral loss of the
enamel samples was calculated by accumulating
mean calcium and phosphate loss for the certain
acids.
Preparation and gaining of solutions for the
photometric assays
At pH 2 and 2.3, after 1, 2, 3, 4, and 5 min, 100 ml
were taken from the acidic solutions and replaced
by 100 ml of fresh acid to maintain the volume and to
keep pH constant. This was controlled with a pH
electrode (RadiometerA/S, Copenhagen, Denmark).
The addition of 100 ml in the respective time led to a
dilution of the samples. This dilution was taken into

544

C. Hannig et al.

Figure 1 Mean calcium (A) and phosphate (B) dissolution from enamel specimens during immersion in hydrochloric acid
(HCl) adjusted to pH 2, 2.3 and 3 [nmol/mm2]. S.D.: standard deviation; (&): pH 2; (~): pH 2.3; (^): pH 3.0.

Figure 2 Mean phosphate release from enamel specimens during 5 min incubation in oxalic acid at pH 2, 2.3 and 3. The
phosphate release is given as [nmol/mm2] of enamel surface. S.D.: standard deviation; (&): pH 2; (~): pH 2.3; (^): pH
3.0.

Erosive effects of different acids on bovine enamel

545

Figure 3 Mean phosphate release of enamel immersed in oxalic acid for 5 min, followed by admixture of fresh oxalic
acid (after 5 min) and of hydrochloric acid (after 10 min) to the oxalic acid solution.

Figure 4 Mean calcium release [nmol mm2 min1] from bovine enamel specimens stored for 5 min in different acids of
pH 2.0 (A), 2.3 (B), 3.0 (C), (HCl: hydrochloric acid; AA: acetic acid; LA: lactic acid; MA: maleic acid; TA: tartaric acid; PA:
phosphoric acid). Significantly different data are signed with different letters (p < 0.05). n = 8 enamel specimens per
subgroup (MV
S.D.).

546

C. Hannig et al.

Figure 5 Mean phosphate release [nmol mm2 min1] from bovine enamel specimens stored for 5 min in different acids
of pH 2.0 (A), 2.3 (B), 3.0 (C), (HCl: hydrochloric acid; AA: acetic acid; LA: lactic acid; MA: maleic acid; TA: tartaric acid;
CA: citric acid). Significantly different data are signed with different letters (p < 0.05). n = 8 enamel specimens per
subgroup (MV
S.D.).

Figure 6 Correlation of mineral loss and proton concentration for all acids and pH levels. Mineral loss as calculated by
accumulation of calcium and phosphate release. Correlation coefficient: r = 0.96.

Erosive effects of different acids on bovine enamel

547

Figure 7 Correlation of mineral loss and the amount of titratable acid for all acids and pH levels. Mineral loss as
calculated by accumulation of calcium and phosphate release. Correlation coefficient: r = 0.26.

consideration in later performed calculation of calcium and phosphate concentration in the solutions.
From the volume taken from the solution, 5, 10 or
20 ml were admixed to 100 ml of the calcium test or
to 200 ml of the phosphate assay, respectively. The
amount admixed to the assays depended on the
calcium or phosphate concentration which was to
be expected according to preliminary trials for the
respective acid.
At pH 3, after 1, 2, 3, 4 and 5 min, 1000 ml were
taken from the incubated volume and replaced
immediately by 1000 ml of fresh acid to maintain

volume and pH level. Due to the low ion concentrations, gained samples were evaporated at 80 8C for
5 h and resolved in 100 ml HCl (pH 2). According to
the concentration of Ca and P which was to be
expected, 5, 10 or 20 ml of the solution were added
to calcium or phosphate assays.

Titration
Amount of titratable acid in the different acidic
solutions at pH 2, 2.3 and 3 was determined by
titration with NaOH (1 mol/l) until pH 7 (Table 2).

Figure 8 Erosive alterations caused by 1 min immersion in (a) oxalic acid, (b) lactic acid, (c) acetic acid and (d) citric
acid (all pH 2).

548

C. Hannig et al.

Figure 9 Erosive alterations caused by 1 min immersion in (a) maleic acid, (b) tartaric acid, (c) phosphoric acid and (d)
hydrochloric acid (all pH 2).

Scanning electron microscopic (SEM)

evaluation of erosive effects
For SEM investigation, samples were treated with
the different acids for 5 min (pH 2). Morphology of
etched surfaces was evaluated with SEM (960, Zeiss,
Oberkochen, Germany, 2.4 kV, 20 mA, 270 s). Air
dried samples were sputtered with gold (Fissons
Instruments, Typ SC 510, Uckfield, UK) resulting in
a gold coating of 7.5  1011 m thickness.

Statistics
Data were evaluated with MannWhitney U-test
(Statistica, StatSoft, Hamburg, Germany). Level
of significance was set at p < 0.05. Where appropriate, tests for correlations were carried out with
Excel 5.0 (Microsoft Office 2000, USA).

Results
Kinetics of calcium and phosphate release
from the enamel during exposition to acids
for 15 min at constant pH
Maleic, lactic, acetic, hydrochloric, phosphoric and
tartaric acid led to a linear dissolution of calcium in

a time dependent manner during incubation for 1

5 min at constant pH. One example is given for
hydrochloric acid (Fig. 1A).
For all acids, except for phosphoric acid, a photometric determination of phosphate release was
performed. With exception of oxalic acid, all acids
led to a linear phosphate dissolution in a time and pH
dependent manner during short time incubation.
For hydrochloric acid, linear kinetics of phosphate
release are depicted exemplarily (Fig. 1B).
Oxalic acid featured a special characteristic
when incubated with enamel specimens. Determination of erosive phosphate dissolution yielded nonlinear kinetics for pH 2 and 2.3 during incubation for
15 min (Fig. 2). In order to investigate the reason
for this observation, specimens were incubated in
oxalic acid for another 5 min. The additional oxalic
acid did not lead to further phosphate dissolution;
phosphate release reached a plateau. After admixture of hydrochloric acid (pH 2) a new increase of
phosphate dissolution was observed (Fig. 3).

Calcium and phosphate release per minute

Linearity of mineral loss during incubation of the
enamel samples for 15 min with the different acids
allowed experimental determination of calcium and
phosphate dissolution per minute (Figs. 4 and 5). For

Erosive effects of different acids on bovine enamel

549

each acid, the erosive calcium and phosphate

release at the different pH levels differed significantly (p < 0.05) (Figs. 4 and 5). At pH 2, lactic acid
caused significantly strongest release of calcium and
phosphate (p < 0.05). At this pH, acetic acid featured weakest erosive effects. At pH 2.3, lactic acid
led to strongest calcium and phosphate loss followed by acetic acid and tartaric acid. Hydrochloric
acid was least erosive. At pH 3, acetic acid was most
erosive compared to all the other acids. Lowest
erosive effects were observed with hydrochloric
acid. In summary, at all pH levels lactic acid was
one of the most erosive acids. Acetic acid was very
erosive at higher pH levels, only.
The ratio of calcium to phosphate release [Ca:P]
ranged between 1.45
0.07 for tartaric acid, pH 3
and 1.94
0.35 for acetic acid, pH 3. Over all acids
and pH values, the ratio was 1.68
0.13 as average.

affected. This is an advantage of the method compared to other methods for evaluation of erosive
effects such as profilometry or determination of
microhardness.13,18,27
In previous studies, amount of mineral dissolved
by erosive drinks, acids, juices, and foodstuffs was
shown to be associated with their titratable amount
of acid, exposure time, temperature, concentration, character of the certain acid and pH under
different test conditions.19,2831 However, to our
knowledge, the present study is the first systematic
attempt to elucidate erosive character of different
pure acids at three certain pH levels in detail via
photometric determination of Ca and P loss during
short time incubation.
Bovine teeth were used as a substitute for human
enamel as done in other investigations.32,33 Enamel
samples were incubated with an excess of acid
providing constant pH levels. This resembles the
situation during consumption of acidic drinks. After
consumption of a low pH drink, the pH on the tongue
and tooth surfaces stays low for about 2 min.14,23 pH
levels between 2 and 3 were chosen as many acidic
drinks range in this level.2,11 In other studies comparing erosive effects of different acids incubation
was performed with acidic solutions of same concentrations.13,27 Due to the different levels of dissociation this will of course lead to different pH
values. In the present study, for direct comparison of
different acids, three standardized pH levels were
chosen, since various beverages of same pH often
contain different acids.
Under the given conditions, there was a clear
influence of pH and type of acid on erosive effects.
The different erosive character of the acids found
for Ca and P dissolution was illustrated by the
scanning electron microscopic evaluation of acid
treated enamel samples. A strong correlation
between mineral loss and proton concentration
was found for all acids. Correlation of pH and erosive
effects has already been proven in other studies by
profilometry. However, in these studies samples
were eroded for 10 or 30 min.13,27
In a profilometric study with 1030 min incubation of the enamel samples, strongest erosions
were observed with PA, less erosive effects were
caused by CA, LA and MA.13 It is noteworthy that in
this experimental setup acidic solutions of same
acid concentrations but of different pH levels were
used for characterization of erosive capacity of the
certain acids. Due to the differing test conditions,
it is not unexpected that grading of the erosive
capacity of the certain acids is different from the
present results where lactic acid and acetic acid
were found to have strongest erosive impact, HCL
was least erosive. In contrast to the present data

Correlation of mineral loss with titratable

acid and proton concentration
There was a strong correlation of the mineral loss
and the proton concentration which is determined
by pH (r = 0.96, Fig. 6). Also, correlation of the
amount of titratable acid (Table 2) and the mineral
loss was calculated (Fig. 7). Only a weak correlation
was recorded (r = 0.28) under the given conditions.
Therefore, it can be concluded that erosive impact
of acids on bovine enamel is predominantly determined by the pH of the acid during short time
exposition at constant pH level.

Scanning electron microscopic evaluation of

erosive effects
All acids caused distinct erosive alterations of the
enamel surfaces. Electron microscopic pictures are
given in Figs. 8 and 9. Oxalic acid yielded precipitations on the enamel surface (Fig. 8a).

Discussion
In the present study, a photometric method for
direct sensitive and specific determination of calcium and phosphate loss during erosive attacks was
adopted.25,26 This allowed to investigate erosive
characteristics of certain acids during short time
incubation in detail. With this method, even minimal erosive effects occurring within 1 min can be
analysed quantitatively. Therefore, it seems to be
acceptable to talk about nano erosive effects
because nmol or ng of Ca and P dissoluted from
enamel can be detected precisely.26 Remineralisable but softened areas in the eroded lesions are not

550

using pure acids, Bartlett et al. found that gastric

juice, which contains mainly HCl was more erosive
than acidic carbonated beverages usually made
with phosphoric or citric acid.4 Under different
conditions also, pure HCL was more erosive than
phosphoric acid.27
One possible reason for the different erosive
effects caused by the acids are specific interactions
of organic acids with hydroxyapatite.34,35 Mono-, di, and tri-carboxylic acids are chemisorbed and
bonded to hydroxyapatite via ionic interactions.34
Thereby, decalcification of and chemisorption to
hydroxyapatite depend on the solubility of the carboxylic salt in its own acidic solution. These chemisorption processes are not influenced by pH or
concentration of the acid. Carboxylic groups are
assumed to substitute hydroxy or phosphate groups
on the surface of enamel or hydroxyapatite, respectively.34,35 Due to the different structures of the
organic acids, different behavior was observed for
oxalic, lactic, citric, and maleic acid. Oxalic acid
was chemically bonded to hydroxyapatite, whereas
maleic, lactic, and citric acid decalcify hydroxyapatite after adsorbing chemically.34,35
Apart from pH or type of acids present in acidic
drinks, also titratable amount of acid is assumed to
have a strong influence on the erosive capacity.2,28
The total acid level as determined via the amount of
titratable acid is considered even more important
than pH of acidic solutions, since it determines the
actual amount of protons available to interact with
the tooth surface.3 However, under the chosen conditions with abundance of acid, the amount of
titratabe acid was not well correlated with dissolution of calcium and phosphate during short time
exposition at constant pH.
Other investigations were based on longtime
exposition of enamel specimens to acidic
reagents.36,37 It should be noted that if a limited
amount of acid is present, erosive effects of the
strong acids are distinctly restricted due to their
high grade of dissociation. Weak acids with high
amount of titratable acid and low level of dissociation have the capability for further dissociation
releasing additional protons if acid is consumed.
In the present study, which was based on a photometric assay, determination of calcium dissolution
induced by oxalic acid and citric acid was not possible. A potential reason for this may be their capacity to form chelating complexes with calcium.38
These chelating processes are influenced by pH and
tend to increase as pH rises since the metal ions
compete directly with hydrogen ions. The chelation
of calcium by oxalate and citrate may detract ions
from the chromogenous molecules applied in the
photometric test.

C. Hannig et al.

SEM analysis showed formation of a precipitate on

the surface of the samples treated with oxalic acid.
It is known that oxalic acid forms precipitates with
calcium.3941 If enamel specimens are treated with
oxalic acid, initial dissolution of enamel is followed
by deposition of calcium oxalate crystals (mono- and
di-hydrate forms) as shown in a previous electron
microscopic study.41 This calcium oxalate layer was
found to prevent further dissolution of phosphate by
oxalic acid, but offers no protection against erosive
attacks by other acids: the addition of HCl increased
phosphate loss from the samples pretreated with
oxalic acid.
Dental enamel consists of about 95 wt.% inorganic mineral which is predominantly characterized
as calcium-phosphate crystals. Hydroxyapatite is
dissolved according to the chemical reaction
Ca10(PO4)6(OH)2 + 8H+ $ 10Ca2+ + 6HPO42 + 2H22H2O.42 The ratio of calcium to phosphate in average amounted to about 1.7 which was confirmed by
the present experimental results underlining precision of the photometric assays applied.42
In interpretation of the present data it must be
regarded that they are basic in vitro results. In the in
vivo situation, the effects of acids on dental hard
tissue are modulated by several factors, such as
other components of acidic beverages like buffering
agents, fluoride, calcium and phosphate.2,29 Also,
salivary flow rate,43 mode of drinking2,3 and especially the aquired salivary pellicle have great impact
on erosive effects.19,44,45 These aspects were
neglected in order to establish a method for numerical description of nano erosion via loss of Ca and P
per mm2 under standardized and reproducible conditions. Further investigations are necessary to elucidate impact of other factors, such as pellicle on
erosive Ca and P dissolution induced by different
acids. Also, mineral content of the surrounding
solution and fluoride content of the enamel specimens may influence the erosive effects. Therefore,
superficial enamel layers containing 20-fold more
fluoride than deeper layers46 were removed in the
standardized grinding procedure to minimize possible fluoride effects.
However, knowledge of the distinct erosive
potential of different acids given as calcium and
phosphate release and their concentrations in beverages allows to use them as variables in a multiple
regression calculating erosive potential of beverages and foodstuffs.28

Conclusion
 The highly sensitive approach used in the present
study allows investigation of minimal erosive

Erosive effects of different acids on bovine enamel

551

effects during short time exposition of enamel to

erosive substrates via photometric determination
of Ca and P loss.
 Different acids expose different erosive character
on enamel as shown via Ca and P release.
 During short time exposition of enamel to various
acids at constant pH, erosive capacity is mainly
determined by pH and type of acid, but not by
amount of titratable acid or concentration of the
certain acid.

20. Lupi-Pegurier L, Muller M, Leforestier E, Bertrand MF, Bolla M.

In vitro action of Bordeaux red wine on the microhardness of
human dental enamel. Arch Oral Biol 2003;48:1415.
21. Grenby TH, Mistry M, Desai T. Potential dental effects of
infants fruit drinks studied in vitro. Am J Nutr 1990;64:273
83.
22. Meurman JH, Harkonen M, Naveri H, Koskinen J, Torkko H,
Rytomaa I, et al. Experimental sports drinks with minimal
dental erosion effect. Scand J Dent Res 1990;98:1208.
23. Millward A, Shaw L, Harrington E, Smith AJ. Continuous
monitoring of salivary flow rate and pH at the surface of
the dentition following consumption of acidic beverages.
Caries Res 1997;31:449.
24. Klimek J, Hellwig E, Ahrens G. Fluoride taken up by plaque,
by the underlying enamel and by clean enamel from three
fluoride compounds in vitro. Caries Res 1982;16:15661.
25. Smith HG, Bauer PJ. Light-induced permeability changes in
sonicated bovine disks: arsenazo III and flow system measurements. Biochemistry 1979;18:506773.
26. Attin T, Becker K, Hannig C, Buchalla WR, Hilgers R. Method to
detect minimal amounts of calcium dissolved in acidic solutions. Caries Res. 2005; in press.
27. West NX, Hughes JA, Addy M. The effect of pH on the erosion
of dentine and enamel by dietary acids in vitro. J Oral
Rehabil 2001;28:8604.
28. Lussi A, Jaggi T, Scharer S. The influence of different factors
on in vitro enamel erosion. Caries Res 1993;27:38793.
29. Featherstone JD, Rodgers BE. Effect of acetic, lactic and
other organic acids on the formation of artificial carious
lesions. Caries Res 1981;15:37785.
30. Cairns AM, Watson M, Creanor SL, Foye RH. The pH
and titratable acidity of a range of diluting drinks and
their potential effect on dental erosion. J Dent 2002;
30:3137.
31. Larsen MJ, Nyvad B. Enamel erosion by some soft drinks and
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