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Determination of Fluoride in Water by Reversed-Phase High-Performance

Liquid Chromatography using F --La 3+-Alizarin Complexone Ternary
Complex

Article  in  Chromatographia · January 2004

DOI: 10.1365/s10337-004-0312-y

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Determination of Fluoride in Water
by Reversed-Phase High-Performance
Liquid Chromatography using F)-La3+-
Alizarin Complexone Ternary Complex
2004, 59, 745–747

X. R. Xu1, H. B. Li1, J.-D. Gu1, 2, 3,&

, K. J. Paeng4
1
Laboratory of Environmental Toxicology, Department of Ecology & Biodiversity, The University of Hong Kong, Pokfulam Road,
Hong Kong SAR, People’s Republic of China
2
The Swire Institute of Marine Science, The University of Hong Kong, Shek O, Cape d’Aguilar, Hong Kong SAR,
People’s Republic of China; E-Mail: jdgu@hkucc.hku.hk (J.-D. Gu)
3
Environmental and Molecular Microbiology, Key Laboratory of Tropical Marine Environment Dynamics (LED),
South China Sea Institute of Oceanography, Chinese Academy of Sciences, 164 West Xingang Road, Guangzhou 510301, P.R. China
4
Department of Chemistry, Yonsei University, Wonju 220-710, South Korea.

Received: 31 October 2003 / Revised: 13 January 2004 / Accepted: 9 February 2004

Online publication: 21 May 2004

resins, it tends to co-elute with short

Abstract retention times with co-existing cations
and anions and its peak also tends to
A method for the determination of fluoride by reversed-phase, high-performance liquid chro- overlap with the water dip negative peak
matography (RP-HPLC) is described. Fluoride, La3+ and alizarin complexone form F-La3+- [9, 10]. On the other hand, HPLC inor-
alizarin complexone ternary complex, which is separated from the matrix on a RP, Ultrasphere ganic ion determination as its complex
C18 column (250 · 4.6 mm, 5 lm) using methanol-water (19:81, v/v) mobile phase at by as an alternative method, has been
1.00 mL min)1; detection at 568 nm. The calibration graph was linear from 1.0–150 ng mL)1 used in the determination of inorganic
for fluoride with a correlation coefficient: 0.9993 (n ¼ 6). The detection limit was 0.2 ng mL)1. ions [11–18], but few examples have
The method was successfully applied to the determination of fluoride in river and tap water. appeared in the literature concerning
Recovery was: 94–102%, RSD in the range: 1.9 –3.6%. non-metallic anions. Although fluoride
can be determined spectrophotometri-
cally as the F)-La3+-alizarin complex-
one, ternary complex [19], the sensitivity
Keywords
and precision of the method are limited
Column liquid chromatography because of overlap of the absorption
Fluoride – alizarin complex bands of the La3+-alizarin complexone
Fluoride in river and tap water complex and the F)-La3+-alizarin com-
plexone ternary complex [5]. If the ter-
nary complex is separated from the
reagent blank and interfering substances
in the sample, the sensitivity and selec-
Introduction determination of fluoride. Spectropho- tivity of the method will be effectively
tometry and the fluoride-ion selective improved. Here we report a novel
Fluoride is an essential trace element; electrode have low sensitivity, for method for determination of fluoride by
low fluoride intakes can cause decayed example, the colorimetric detection lim- RPLC as the F)-La3+-alizarin com-
teeth and is toxic at high concentrations. its for fluoride are rarely better than plexone ternary complex. The proposed
High fluoride intakes can also cause 0.020 lg mL)1, and it is frequently rec- method was about 100-times more sen-
mottling of teeth and bone disorders, ommended that the fluoride-ion selective sitive than the spectrophotometric one
hence it has received much attention electrode should not be used for fluoride using the F)-La3+-alizarin complexone
with regard to medicinal, pharmaceuti- concentrations < 0.1 lg mL)1 because ternary complex [19]. The present meth-
cal, public health and environmental of the risk of erratic electrode response od was more sensitive than previous
protection [1, 2]. [8]. Fluorimetry is more sensitive than HPLC ones [18]. Sensitivity was also
In the literature, several methods, potentiometry and spectrophotometry. comparable with ion chromatographic
including potentiometry (ion selective Generally, however, fluoride must be methods [8, 9] and was successfully
electrode) [3], spectrophotometry [1, 4, separated prior to determination to applied to determine fluoride in river
5], fluorimetry [6, 7], and ion chroma- avoid interference [6]. As fluoride is very water and tap water.
tography [8, 9], have been proposed for weakly retained by anion-exchange

Short Communication Chromatographia 2004, 59, June (No. 11/12) 745

DOI: 10.1365/s10337-004-0312-y
0009-5893/04/06
2004 Friedr. Vieweg & Sohn Verlagsgesellschaft mbH
 (a)
0.012
0.009
Absorbance (au)
0.006
0.003
0 0
0 2 4 6
Retention time (min)
Fig. 1. Chromatograms of fluoride in standard solution (A) and river water (B): 1 ¼ F)-La3+-alizarin complexone ternary complex
Experimental
Apparatus
The LC system used throughout consisted
of two Waters 510 pumps (Waters, Mil-
ford, MA, USA), a sample injector (Rhe-
odyne, Cotati, CA, USA) with 20 lL loop
and a Waters 996 photodiode array
detector. Evaluation and quantification
were on a Millennium Chromatography
data system (Waters, Milford, MA, USA).
The column was a reversed-phase Ultra-
sphere C18 column (250 · 4.6 mm I.D.,
5 lm, Beckman, Fullerton, CA, USA).
Peak areas were used for quantification.
Reagents
All solutions were prepared with AR
grade compounds. Reverse osmosis-Milli
Q water (18 MW) (Millipore Corp., USA)
was used for all solutions and dilutions.
Stock fluoride solution: 1.0 mg mL)1
(prepared by dissolving 0.2210 g sodium
fluoride [Sigma, USA] in 100 mL water)
was stored in a polyethylene bottle. A
working standard solution was prepared
by suitable dilution of stock solution with
water. The alizarin complexone (BDH,
UK) solution was 0.002 M, and the lan-
thanum nitrate (Sigma, USA) solution
was 0.002 M. Acetate buffer solution was
prepared by addition of 0.1 M sodium
acetate (Sigma, USA) solution to
0.100 M acetic acid (BDH, UK) until the
pH ¼ 4.3. The composite fluoride reagent
comprised the alizarin complexone, lan-
thanum nitrate and acetate buffer solu-
tions as well as acetone (2:2:1:4, v/v) [19].
All other solutions were prepared by
dissolving appropriate amounts of com-
mercially available chemicals in water.
746
0.04
0.03
Absorbance (au)
0.02
1
0.01
8 10 0 2
Procedures
All chromatographic separations were
carried out at ambient temperature. The
mobile phase was methanol-water (19:81,
v/v) and the flow-rate: 1.00 mL min)1.
The injection volume was 20 lL; the
column eluate was monitored at 568 nm;
the mobile phase was filtered through a
0.45 lm membrane and degassed by
sonication prior to use.
A certain (moderate) volume of stan-
dard fluoride solution was transferred to a
10 mL volumetric flask and 4.00 mL of the
composite fluoride reagent added and well
mixed. After 10 min, 1.00 mL triethyl-
amine was added and, finally, the solution
was diluted to the mark with water.
A 2.00 mL water sample was trans-
ferred to a 10 mL volumetric flask, and
the other procedures were the same as for
the standard sodium fluoride solution
described above. The sample solution was
also filtered through a 0.45 lm mem-
brane filter before injection.
Results and Discussion
F), La3+ and alizarin complexone form
F)-La3+-alizarin complexone ternary
complex [1, 5, 19], which may be deter-
mined by spectrophotometrically. How-
ever, the sensitivity and precision of this
method are limited because of overlap of
the absorption bands of the La3+-alizarin
complexone complex and the F)-La3+-
alizarin complexone ternary complex [5].
To separate the ternary complex and the
reagent blank, an LC method was adop-
ted. In a preliminary study, although the
F)-La3+-alizarin complexone ternary
complex was stable for up to 1 h in
solution, it was unstable on a C18 column
Chromatographia 2004, 59, June (No. 11/12)
(b)
1
4 6 8 10
Retention time (min)
and the ternary complex peak could not
be obtained. The complex became very
stable on the C18 column after triethyl-
amine was added to the solution of the
ternary complex. If triethylamine was
added to the composite fluoride reagent
before adding fluoride, it would be very
difficult to form the F)-La3+-alizarin
complexone ternary complex [19]. Tri-
ethylamine should therefore be added
after reacting fluoride with the composite
fluoride reagent for 10 min: when the
analyte solution may be injected imme-
diately. The complex was stable in the
solution and on a C18 column for up to
2 h at room temperature, and fluoride ion
in the sample solution was determined in
the form of the F)-La3+-alizarin com-
plexone ternary complex by LC. Fur-
thermore, the reagent blank and matrix
interference could be separated from the
ternary complex on a C18 column with
methanol-water mobile phase (19:81, v/v).
The effect of methanol in the mobile
phase on separation of the ternary com-
plex and the La3+-alizarin complexone
complex was significant. When methanol
in the mobile phase was >25%, the ter-
nary complex could not be separated
from the La3+-alizarin complexone
complex. However, when methanol in
mobile phase was <15%, retention of the
ternary complex was >15 min, and the
peak broadened. Methanol-water (19:81,
v/v) was therefore chosen as mobile
phase: flow-rate: 1.00 mL min)1; detec-
tion wavelength: 568 nm – which was the
maximum absorption wavelength of the
complex between 400–700 nm in the
mobile phase. Under these conditions,
representative chromatograms of fluoride
in standard solution and river water are
shown in Figs. 1A and B, respectively.
The chromatogram of fluoride in tap
Short Communication

water was similar to that of river water
(data not shown).
The calibration graph was linear from
1– 150 ng mL)1 for fluoride with a cor-
relation coefficient of 0.9993 (n ¼ 6). The
linear regression equation is given by:
A ¼ 195 C ( ng mL)1) + 850, where A is
the peak area of the ternary complex,
while C is the concentration of fluoride.
The RSDs of the slope and intercept were
2.7% and 3.1%, respectively. The detec-
tion limit (S/N ¼ 3 [20]) was 0.2 ng mL)1.
The RSD was 2.5% for determination of
30 ng mL)1 fluoride standard (n ¼ 8).
The stability of the method was good and
the RSD was < 5% for 30 ng mL)1
fluoride determined on seven consecutive
days (once a day).
The interference of a number of dif-
ferent ions was studied by spiking
20 ng mL)1 fluoride samples with known
quantities of foreign materials and anal-
ysing them by the present method. Under
the present conditions of determination,
no interference (relative error <±5%)
was observed at ratios (m:m) of 1000:1
) ) ) 47:547–559
for K+, Na+, NHþ

4 , Cl , Br , I , NO3 ,
2+
HCO3 , CO3 , and Ca ; 100/1 for NO


2

2,
Mg2+, SO2
2

4 ; and SiO3 . There was some
interference of Cu2+, Zn2+, Fe3+ and
Al3+ at ratios m:m of 10:1. For some
kinds of water samples with high con-
centrations of interfering ions, the cations
could be removed by passing the sample
through a cation-exchange column
(Dowex 50W-X8, H+ form) [9], before
pre-column derivatization (complexa-
tion).
The present method was applied to
determine fluoride in real samples, i.e.,
river water and tap water. Results are
given in Table 1 and as shown there,
recovery was 94–102% and the RSD was
in the range: 1.9–3.6%. The results
agreed with those obtained by the stan-
dard method (ion-selective electrode)
[21]. The t-test proved that no significant
difference between the present and the
standard methods could be observed at
the 95% probability level. Three spectra:
200–700 nm were taken from the apex,
upslope and downslope points from the
peak of the ternary complex in the chro-
matogram. They completely overlapped
each other, which indicated that the peak
of the ternary complex was pure. The
fluoride in a certified reference material of
human urine (GBW 09106) was also
determined by this method. The certified
value of the urine sample was 0.62 ±
0.04 lg mL)1. The result obtained by the
Short Communication
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a
Table 1. Determination of fluoride in river water and tap water
Samples Fluoride RSDb Fluoride Recovery Fluoride
content (%) added (%) contentc
(lg mL)1) (lg mL)1) (lg mL)1)
River water 1 0.21 2.3 0.25 97 0.22
River water 2 0.27 1.9 0.25 94 0.25
River water 3 0.18 3.1 0.25 101 0.19
Tap water 1 0.23 2.1 0.25 98 0.24
Tap water 2 0.16 3.6 0.25 102 0.15
Tap water 3 0.19 2.7 0.25 96 0.21
a
Average of three determinations.
b
n=8.
c
Results by standard method [21].
present method was 0.59 lg mL)1 with the determination of fluoride in river wa-
an RSD (n ¼ 8) of 2.3%. ter and tap water. It can be employed as
Compared with the spectrophotomet- an alternative analytical method for the
ric method [4, 19], the proposed method determination of trace fluoride.
was more sensitive, and detection limits
were 20 ng mL)1 [4], 20 ng mL)1 [19],
and 0.2 ng mL)1 (present method), References
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