DOI: 10.

2478/s11532-006-0042-0
Research article
CEJC 4(4) 2006 798–807

Simultaneous determination of fluoride and

monofluorophosphate in toothpastes by supressed
ion chromatography

Rajmund Michalski∗ , Barbara Mathews

Institute of Environmental Engineering of Polish Academy of Science
41-819 Zabrze, Poland

Received 28 June 2006; accepted 31 July 2006

Abstract: Suppressed ion chromatography with a highly selective and high capacity anion-exchange
column is used for the determination of anionic compounds in ten commercially available toothpastes.
Minimal sample preparation is needed, requiring only dilution and filtration over a solid-phase extraction
cartridge. Quantitative analysis of fluoride and monofluorophosphate, as well as chloride, nitrate,
orthophosphate, and sulfate is achieved in one chromatographic run with a total analysis time of 25
minutes.
Optimized analytical conditions are validated in terms of accuracy, precision, and total uncertainty, and
the results demonstrate the reproducibility of ion chromatography. The coefficient of determination for
the anions ranges from 0.9896 to 0.9997. The spiked recoveries for the anions are 88-108%. The change
in the fluoride content of the toothpaste with storage time is investigated.
c Versita Warsaw and Springer-Verlag Berlin Heidelberg. All rights reserved.

Keywords: Toothpaste, fluoride, monofluorophosphate, ion chromatography

1 Introduction

Fluoride occurs naturally in trace quantities in soil, water, plants, and animals. When
fluoride is ingested, some is taken up by body tissues with long-term deposition in teeth
and bones. The biological role and importance of fluorides revealed by studies in re-
cent decades have determined their use in prophylaxis and the treatment of disease;
small quantities of fluorides and its compounds are added to food, tablets, and especially
toothpastes and some dental gels.
∗
E-mail: michalski@ipis.zabrze.pl

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Toothpastes can be divided into categories of non-fluoridated and fluoridated with

sodium fluoride, sodium monofluorophosphate or aminofluorides. Fluoride toothpastes
were introduced into the market in 1973. Silica-based toothpastes usually contain NaF
and calcium carbonate-based – monofluorophosphate.
Fluoride and its compounds are the active ingredients in toothpastes used to reduce
bacteria-producing plaque. In addition, carbonate and phosphates, such as pyrophos-
phate and tripolyphosphate, are used in toothpaste to reduce tartar. Saccharin is used
as sweetener, and sorbitol and glycerol are used as moisture control agents. Furthermore,
toothpaste contains other compounds such as detergents, flavors, dyes, and preservatives.
The US Food and Drug Administration has approved the use of three decay-preventing
compounds in toothpaste: stannous fluoride, sodium fluoride, and sodium monofluo-
rophosphate (MFP). Fluoride can react with other ingredients in toothpaste to form
insoluble compounds and therefore can be unavailable to fulfill its therapeutic function.
Monofluorophosphate can hydrolyze to free fluoride and phosphate ions during storage.
The determination of fluoride and MFP in toothpaste is important in order to eval-
uate the quality and stability of these cavity-preventing compounds. It is also necessary
to develop analytical procedures not only for the determination of MFP ion, but also
for the simultaneous determination of MFP and its hydrolysis products, fluoride and
orthophosphate.
For the quality control of toothpaste, a fast and powerful technique is required to deter-
mine the quantity of the active ingredients. Several analytical techniques have been used
to determine the fluoride content in dental care products, including gas chromatography
[1], ion-selective electrodes [2], spectrophotometric methods [3], and electrophoresis [4].
Some of these methods suffer from interferences and limited sensitivities; they can
be labour intensive and are often difficult to automate. Moreover, most of them have
the disadvantage that MFP must be hydrolysed in acid first before the total fluoride is
determined. The fluoride content from the acid hydrolysis is difficult to quantify, because
any HF formed during hydrolysis may evaporate.
Ion chromatography is an innovative analytical technique that has significantly im-
proved the analysis of anions and cations. Ion chromatography eliminates the need to
use hazardous reagents, which are often integral to wet chemical methods.
There are many important advantages of using IC: a broad range of applications, well-
developed hardware, many detection options, reliability (good accuracy and precision),
high selectivity, high speed, high separation efficiency, good tolerance to sample matrices,
and a low cost of consumables. In addition, ion chromatography is widely accepted as a
standard, reference methodology for water and wastewater analysis [5].
Gradient ion chromatography has been used for monofluorophosphate determination
in hardened paste [6] and total fluoride content in dentifrices [7]. Furthermore, Dionex
recommends gradient elution for fluoride and monofluorophosphate analysis using a hy-
droxyselective, high capacity column [8]. Although gradient elution is a very useful mode
for complex matrix sample analyses, most ion chromatography users have isocratic sys-
tems with conductivity detectors.

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In this work, the suppressed isocratic ion chromatography method is used for the
simultaneous determination of fluoride, monofluorophosphate and four common anions
(chloride, nitrate, phosphate, and sulphate) found in commercially available toothpastes.
The changes in fluoride ion concentration in toothpaste depending upon the length of stor-
age time are investigated. Ion chromatography and ion-selective electrode potentiometry
are compared for fluoride ion determination.

2 Experimental
Ion chromatographic separations were performed on a Metrohm ion chromatography sys-
tem (Herisau Metrohm AG, Switzerland) equipped with an 818 IC Pump, an 837 IC
Eluent Degasser, an 830 IC Interface, an 820 IC Separation Center, a Rheodyne injec-
tion valve, Metrodata 2.3 software, an autosampler (838 Advanced Sample Processor),
an MSM suppressor, and a conductivity detector (819 IC).
Determination of fluoride ion concentration with ion-selective electrode potentiome-
try was carried out using a CPI-502 IonMeter (Elmetron, Poland) and an ion-selective
electrode, 09-37 from Marat (Marat, Poland).
Analytical grade Na2 CO3 and NaHCO3 used for the eluent preparation and 95%
H2 SO4 used as a regenerant solution were obtained from Fluka (Fluka, Steinheim, Switzer-
land). Standard solutions of inorganic anions were prepared by dissolving of suitable salts
in water. Analytical grade KF, NaCl, NaNO3 , K3 PO4 and Na2 SO4 were purchased from
Fluka (Fluka, Steinheim, Switzerland). Sodium monofluorophosphate was obtained from
Sigma Aldrich (Sigma Aldrich, USA).
Certified reference materials for MFP are not available commercially, therefore a ref-
erence standard solution (AccuIonT M , NIST, USA) containing F− (20 mg L−1 ), Cl− (30
3− 2−
mg L−1 ), NO− −1 −1 −1
3 (100 mg L ), PO4 (150 mg L ) and SO4 (150 mg L ) was used for
the recovery test. Water used in the experiments was purified using Millipore equipment
(Millipore, Bedford, MA, USA) and had an electrical conductivity < 0.05 µS/cm.
Standard solutions of fluoride, MFP, and the other anions (1000 mg/L) were prepared
by dissolving the appropriate amounts of the corresponding salts in deionized water.
Calibration solutions were prepared by diluting the standard solutions to the required
concentration just before use. All of the above solutions were diluted to the proper
concentrations as required and were kept in polyethylene flasks at + 4 ◦ C.

3 Results and discussion

3.1 Sample preparation

Ten different toothpastes produced by international producers (e.g. GlaxoSmithKline,
Palmolive, Procter&Gamble, Unilever) were bought on the market. A 1.000 ± 0.002 g
sample of each toothpaste was dissolved in 10 mL of deonized water. The samples were
stirred, and a 5 mL of suspension of each sample was passed through an OnGuard RP

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(Dionex, USA) solid-phase extraction cartridge in order to remove most of the non-polar
organic compounds. Next, each sample was filtered through a 0.15 µm membrane filter
and injected directly onto the anion-exchange column.

3.2 Method optimization

The separation columns used for the optimization test were a Metrosepp A SUPP 5
(150 × 4 mm, I.D.) from Metrohm (Metrohm, Herisau, Switzerland), an Ion Pac AS9-
SC (250 × 4 mm, I.D.), and anIonPac AS 14 (250 × 4 mm I.D.) from Dionex (Dionex,
Sunnyvale, USA). These anion-exchange columns produce the proper resolution between
fluoride ion and a negative water dip, as well as the resolution of fluoride ion among the
other anions without increasing the total analysis time over reasonable time frame.
Two eluent concentrations (1.7 mM Na2 CO3 + 1.5 mM NaHCO3 and 3.5 mM Na2 CO3
+ 1.0 mM NaHCO3 ) and two eluent flow rates (0.8 mL min−1 ; 1.0 mL min−1 ) were tested
in oder to optimize the separation. The column oven was maintained at 32 ◦ C. Samples
were injected using 20 µL loop injector.
The optimum chromatographic performance occurs using the Dionex IonPac AS 14
column preceded by an IonPac AG 14 guard column using the 3.5 mM Na2 CO3 + 1.0
mM NaHCO3 eluent with a flow rate of 0.8 ml min−1 .

3.3 Calibration and method validation

Validation based on ISO guidelines [9] was carried out with responses for peaks areas
collected and evaluated. Ten calibration solutions of inorganic anions were injected into
the eluent stream. The concentrations ranges were chosen according to the expected
concentration of the anions in the solutions prepared from the toothpaste samples. All
calibration solutions were analysed in triplicate under the optimized chromatographic
conditions.

Table 1 Method validation parameters.

Parameter F− Cl− NO−

3 MFP PO3−
4 SO2−
4

Concentration range [mg L−1 ] 1-10 0.5-5 0.5-5 10-100 10 - 100 10 - 100
Standard deviation [mg L−1 ] 0.60 0.52 0.87 1.36 1.47 1.47
Coefficient of variation [%] 2.35 1.39 0.85 2.78 3.63 3.63
Limit of detection [mg L−1 ] 0.15 0.11 0.18 0.34 1.14 1.07
Limit of quantification [mg L−1 ] 0.45 0.33 0.54 1.02 3.42 3.21
Regression coefficient (r) 0.9981 0.9987 0.9968 0.9896 0.9974 0.9982

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Table 2 Concentrations of fluoride and selected inorganic anions in toothpaste water solutions (n=3) [mg L−1 ].

Electrical F− Producer data

Toothpaste pH conductivity Ion Ion-selective Cl− NO−
3 MFP PO3−
4 SO2−
4 fluoride compounds
[µS cm−1 ] chromatography potentiometry content

1 7.23 167.70 8.13 ± 0.16 7.89 ± 0.22 1.07 ± 0.09 0.37 ± 0.03 < LOD 0.95 ± 0.05 57.88 ± 1.43 NaF 0.31%
2 7.49 266.80 3.59 ± 0.05 1.54 ± 0.06 0.78 ± 0.02 0.40 ± 0.02 90.42 ± 1.67 319.33 ± 18.3 3.92 ± 0.11 NaF 0.1%, MFP 0.76%
3 6.85 252.20 8.17 ± 0.19 7.81 ± 0.19 0.38 ± 0.02 0.42 ± 0.02 < LOD 156.26 ± 9.52 41.06 ± 1.26 NaF 0.32%
4 7.84 716.62 1.76 ± 0.08 0.85 ± 0.06 0.24 ± 0.02 0.38 ± 0.02 75.90 ± 1.23 29.57 ± 1.05 31.00 ± 0.95 NaF 0.1%, MFP 0.76%
5 6.63 328.67 1.47 ± 0.06 1.27 ± 0.08 0.61 ± 0.03 0.45 ± 0.03 < LOD 4.25 ± 0.15 41.19 ± 1.01 Sodium aminofluoride 0.125%

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6 7.08 558.29 9.27 ± 0.19 8.27 ± 0.21 0.46 ± 0.03 0.28 ± 0.02 < LOD 234.60 ± 13.3 51.28 ± 1.38 NaF 0.25%
7 7.41 614.56 7.99 ± 0.17 7.45 ± 0.22 1.74 ± -0.08 0.41 ± 0.03 < LOD 37.35 ± 1.37 36.36 ± 1.29 NaF 0.31%

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8 7.62 662.07 7.76 ± 0.15 7.17 ± 0.19 0.24 ± 0.02 0.32 ± 0.02 < LOD 51.29 ± 1.44 50.26 ± 1.34 NaF 0.36%
9 6.84 514.72 2.62 ± 0.08 2.34 ± 0.07 0.32 ± 0.02 0.52 ± 0.04 < LOD 1.29 ± 0.09 29.54 ± 0.95 Sodium aminofluoride 0.125%
10 7.17 508.33 8.26 ± 0.20 6.15 ± 0.13 0.27 ± 0.02 0.72 ± 0.05 66.62 ± 1.19 49.28 ± 1.57 41.93 ± 1.15 NaF 0.15%, MFP 0.76%
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Concentration ranges of the anions in the calibration solutions, standard deviations,

coefficients of variation, limits of detection, limits of quantification and regression coeffi-
cients are given in Table 1. The limit of detection (LOD) and the limit of quantification
(LOQ) were calculated as six and ten times the ratio between the standard deviation of
the regression and the slope of the calibration line, respectively.
The pH value, electrical conductivity and mean (n=3) concentrations of the anions
in the solutions prepared from the toothpaste samples (including standard deviation) are
given in Table 2. Because of the high content of PO3− 4 ions, samples no.2, 4, and 6 were
diluted with deonized water (1:10 v/v) before injection. Moreover in Table 2, the results
obtained for fluoride ion using ion chromatography and ion-selective potentiometry meth-
ods are compared. The table also contains producer data about the fluoride compounds
and content in the toothpastes. A sample chromatogram obtained for sample no.4 is
shown on Figure 1.

mV

400 MFP 75.82

350

300
Fluoride 1.72
250

200

150
Sulfate 31.15

100 Phosphate 29.74

Chloride 0.21
50
Undefined peak Nitrate 0.35

0
ch1

0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 min

Fig. 1 An example chromatogram of sample no. 4.

The recovery study was performed by adding 1.0 mL of Reference Standard Solution
(AccuIonT M , NIST, USA) to 1.0 g of toothpaste and 9.0 mL of deonized water. The
whole mixture was stirred and after purification was analysed chromatographically. The
recoveries of the analyzed anions are listed in Table 3.

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Table 3 Recovery of fluoride and common inorganic anions from analysed samples.
F− Cl− NO−
3 PO3−
4 SO2−
4
Toothpaste Added Found Recovery Added Found Recovery Added Found Recovery Added Found Recovery Added Found Recovery
[mg L−1 ] [%] [mg L−1 ] [%] [mg L−1 ] [%] [mg L−1 ] [%] [mg L−1 ] [%]

1 + 0.47 94 + 0.72 96 + 2.31 92 + 3.69 98 + 3.62 96

2 + 0.55 102 + 0.82 109 + 2.69 108 + 3.82 102 + 3.78 101
3 + 0.53 106 + 0.70 93 + 2.53 101 + 3.45 92 + 3.65 97
4 + 0.55 102 + 0.84 102 + 2.67 107 + 3.81 102 + 3.49 93
5 0.50 + 0.49 98 0.75 + 0.69 92 2.50 + 2.39 96 3.75 + 3.55 95 3.75 + 3.29 88

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6 + 0.52 104 + 0.66 88 + 2.32 93 + 3.32 88 + 3.62 96
7 + 0.47 94 + 0.71 95 + 2.27 91 + 3.84 102 + 3.28 87

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8 + 0.46 92 + 0.77 103 + 2.37 95 + 3.52 94 + 3.59 96
9 + 0.51 102 + 0.73 97 + 2.45 98 + 66 97 + 3.51 94
10 + 0.54 108 + 0.81 108 + 2.64 106 + 3.93 105 + 3,84 102
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3.4 The influence of toothpaste storage time on the content of fluoride

ions
During storage, MFP hydrolyses to phosphate and fluoride ions. On the other hand free
fluoride can react with other ingredients in toothpastes to form insoluble compounds. In
order to study these effects, an analysis of the toothpaste aging was carried out. All
toothpastes were analyzed 4 times: first just after purchasing (batch “0”), and then after
one (batch “1”), two (batch “2”) and three (batch “3”) months. The changes in the
concentration of fluoride and MFP are shown on Figure 2.

12

Batch ”0” Batch “1”

Batch “2” Batch “3”

10

8
Fluoride [mg/L]

0
1 2 3 4 5 6 7 8 9 10
Sample

Fig. 2 The changes of fluoride content in toothpastes during 3 month storage.

According to the results obtained in this study, the following can be concluded.
(1) Suppressed isocratic ion chromatography using an anion-exchange column, a car-
bonate/bicarbonate eluent, and a conductivity detector was validated and applied
3− 2−
to the determination of F− , MFP, Cl− , NO− 3 , PO4 and SO4 in toothpastes. This
method is an alternative, simple, robust, and low cost method compared to wet
methods used for inorganic ion analysis. Suitable selection of analytical conditions
(column, eluent, flow rate) produced results comparable with more sophisticated
gradient elution.
(2) Validation parameters show that this method is accurate, sensitive, and reliable
under the described analytical conditions. The calculated percentage recovery is in
the range of 88-108%.

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(3) The mean content of fluoride ions in the samples ranges from 1 – 10 mg/L. According
to the producers’ data, the MFP was detected in only three of the ten samples.
(4) The concentrations of chloride and nitrate were equivalent to the background of the
water used for sample preparation. The concentration of sulfate originating from the
toothpaste ingredients does not exceed 60 mg/L. The content of phosphate does not
correlate with the presence of MFP in toothpastes. According to the producers’ data,
its presence originates from other compounds added to toothpastes (e.g. potassium
phosphate, sodium pyrophosphate).
(5) There is no significant difference in the concentration of fluoride ions in the tooth-
pastes during their three month storage, although its concentration decreases slightly.
Thus, the content of available fluoride ions is stable and does not appear to depend
upon a reasonable storage time.
(6) According to the literature [10], the differences between the fluoride concentrations as
determined by ion chromatography and ion-selective potentiometry can be explained
by considering the presence of some metals in samples (e.g. aluminium).
(7) For future study, the influence of storage conditions (e.g. temperature and long-term
storage) should be carried out.

References

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[8] Dionex AN 156, Determination of Anions in Toothpaste by Ion Chromatography,

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