# Rapid and Sensitive Determination of Trace Chloride Ion in Drinks Using Resonance Light Scattering Technique

Authors: Hui Cao; Dong Hui Wu
Journal: Journal of Automated Methods and Management in Chemistry
 (2008)

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## Abstract

A resonance light scattering (RLS) technique to determine chloride ions in drinks was developed. Chloride ions were found to bindAg+ forming AgCl aggregates that produced intense resonance scattering light. Effects of factors such as acidity, ionic strength, and coexistent interferents on the RLS of AgCl aggregates were investigated. The pH of solution almost did not affect the production of RLS and few foreign species interfered with the detection of chloride ions. The resonance scattering light intensity at the maximum peak of 571 nm was linear to the concentration of chloride ions in the range of 1.42–8.52 ng mL-1 with a detection limit of 0.71 ng mL-1. To determine the feasibility of the proposed method, some samples of water and drinks were analyzed. The attained results were in agreement with that of ion-selective electrode method. Good recovery results were also obtained with the range of 94.08–105.63%. The sensitivity and selectivity of the RLS method are high enough to determine trace amounts of chloride ions without any significant interference from high concentration of other components such as common anions and cations.


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## Body

## 1. Introduction

Resonance light scattering (RLS) is
an elastic scattering and occurs when an incident beam in energy is close to an
absorption band. Pasternack et al. first established the RLS technique to study
the biological macromolecules by means of an ordinary fluorescence spectrometer
[1]. Due to its high sensitivity, selectivity, and convenience, RLS studies
have attracted great interest among researchers. In recent years, RLS technique
has been used to determine pharmaceutical [2–5], ion
[6], bacteria [7], and various biological macromolecules, such as nucleic acids [8–10], proteins [11–15], and peptide
[16].Over the years, numerous analytical methods for Cl−  in a variety of samples have been developed, such as ion chromatography
[17, 18], near-infrared spectrometry [19],
spectroscopy [20], ion-selective electrode method [21], turbidimetric
method [22], and so on. Among these methods, the turbidimetric method was
popular and regarded relatively reliable for the quantification of Cl−. Although it often
provided very accurate results, it suffered from the long experimental time, lower
sensitivity, and complexity. Partly, because of the lower sensitivity, few
of the above methods were applied to determine Cl− in biological
systems. The classical ion-selective electrode method is currently used in food
analysis of Cl− in drinks. Nevertheless, these
methods similarly suffered from the drawbacks of low sensitivity and
inapplicability to low chloride-containing samples.The objectives of this report are to establish a rapid, sensitive, and selective RLS analytical method
to determine of Cl− and to develop an alternative standard method for determination of Cl−  in marketing
drink. The obtained results were compared with that of the ion-selective
electrode method.


## 2. Experimental

### 2.1. Apparatus

RLS spectra were recorded on a JASCO FP-6500 spectrofluorophotometer equipped with a thermostated cell
compartment using quartz cuvettes (1.0 cm) (Japan). The width of excitation and
emission slits was set at 3.0 nm. A Bayer Rapidchem 744 electrolyte
analyzer (UK) was used in the ion-selective electrode detection. The pH measurements were carried out on a
PHS-3C Exact Digital pH meter equipped with Phonix Ag-AgCl reference electrode
(Cole-Parmer Instrument Co., Ill, USA), which was calibrated with standard
pH buffer solutions.


### 2.2. Reagents

A stock solution containing 355.00 ng mL−1 of Cl− was prepared by
dissolving 0.5850 g of NaCl (>99.99%, Sigma Co., Mo, USA) in doubly distilled water and diluting to
1000 mL. The working solutions were obtained by diluting the stock solution
prior to use. A working solution of silver nitrate (169.87 μg mL−1)
was prepared with doubly distilled water. All other reagents and solvents were
of analytical reagent grade and used as purchased. Double-distilled water was
used throughout. HAc and NH3·H2O (0.01 mol L−1)
were used to control the acidity, while 0.10 mol L−1 NaNO3 was
used to adjust the ionic strength of the aqueous solutions.


### 2.3. Standard Procedure

An appropriate aliquot of Cl− working solution was added to 1.0 mL of silver nitrate working
solution and diluted to 10.0 mL with water. After
standing for five minutes, the solution was synchronous scanned on the spectroflourometer through the
wavelength range of 250–750 nm. The
obtained RLS spectra were recorded, and the intensity was
measured at 571 nm. The enhanced RLS signal of AgCl system was represented as ΔI=I−I0
(I and I0 are the RLS intensity of the system in the presence and absence of Cl−, resp.).
The operations were carried out at 20°C.


### 2.4. Sample

The mineral water, pure
water, and green tea commonly found in the supermarket were used as samples.
The famous brands such as Nongfu spring, Laoshan, Nestlé, Robust, Watsons, and
Masterkong were taken into account. These samples were diluted with
double-distilled water and sterilized by filtration (0.2μm) before testing.
Chloride ion in each sample was determined by the above-mentioned procedures by
adding a 1.0 mL aliquot of the prepared sample instead of the chloride ion working solution.


## 2.1. Apparatus

RLS spectra were recorded on a JASCO FP-6500 spectrofluorophotometer equipped with a thermostated cell
compartment using quartz cuvettes (1.0 cm) (Japan). The width of excitation and
emission slits was set at 3.0 nm. A Bayer Rapidchem 744 electrolyte
analyzer (UK) was used in the ion-selective electrode detection. The pH measurements were carried out on a
PHS-3C Exact Digital pH meter equipped with Phonix Ag-AgCl reference electrode
(Cole-Parmer Instrument Co., Ill, USA), which was calibrated with standard
pH buffer solutions.


## 2.2. Reagents

A stock solution containing 355.00 ng mL−1 of Cl− was prepared by
dissolving 0.5850 g of NaCl (>99.99%, Sigma Co., Mo, USA) in doubly distilled water and diluting to
1000 mL. The working solutions were obtained by diluting the stock solution
prior to use. A working solution of silver nitrate (169.87 μg mL−1)
was prepared with doubly distilled water. All other reagents and solvents were
of analytical reagent grade and used as purchased. Double-distilled water was
used throughout. HAc and NH3·H2O (0.01 mol L−1)
were used to control the acidity, while 0.10 mol L−1 NaNO3 was
used to adjust the ionic strength of the aqueous solutions.


## 2.3. Standard Procedure

An appropriate aliquot of Cl− working solution was added to 1.0 mL of silver nitrate working
solution and diluted to 10.0 mL with water. After
standing for five minutes, the solution was synchronous scanned on the spectroflourometer through the
wavelength range of 250–750 nm. The
obtained RLS spectra were recorded, and the intensity was
measured at 571 nm. The enhanced RLS signal of AgCl system was represented as ΔI=I−I0
(I and I0 are the RLS intensity of the system in the presence and absence of Cl−, resp.).
The operations were carried out at 20°C.


## 2.4. Sample

The mineral water, pure
water, and green tea commonly found in the supermarket were used as samples.
The famous brands such as Nongfu spring, Laoshan, Nestlé, Robust, Watsons, and
Masterkong were taken into account. These samples were diluted with
double-distilled water and sterilized by filtration (0.2μm) before testing.
Chloride ion in each sample was determined by the above-mentioned procedures by
adding a 1.0 mL aliquot of the prepared sample instead of the chloride ion working solution.


## 3. Results and Discussion

### 3.1. Characteristics of the RLS Spectra

The RLS spectra of Cl−  and AgNO3 in water were shown in Figure 1(a) and (b), respectively. It can be seen that
the RLS intensity of AgNO3 solution is quite weak in the whole
scanning wavelength region. In contrast, upon addition of trace amount of Cl− to AgNO3 solution, a remarkably enhanced RLS with a maximum peak at 571 nm was observed
under the same conditions (1.0 mL of silver nitrate). The
addition of increasing Cl−  to the solution
leads to the gradual enhancement in RLS intensity, exhibiting a
concentration-dependent relationship. The production of RLS and its intensity are correlative with the formation of the aggregate and its
particle dimension in solution. Bearing on this point, we
inferred that the Cl− ion may displace
NO3− ion in AgNO3 (aq), forming a new AgCl (s) compound that
could be expected to be an aggregate. The size of AgCl (s)
particles may be much less than the incident wavelength, and thus the enhanced
light-scattering signal occurs under the given conditions. In this way, the resonance
light scattering formula could be applicable to the AgCl
system.Figure 1
RLS profile of
AgCl systems. (b)–(h) 6.99μg mL−1 of AgNO3,
1.0 mg mL−1 tween-80, and 0.0, 1.42, 2.84, 4.26, 5.68, 7.10,
8.52 ng mL−1 of Cl−. (a) 35.5 ng mL−1 of Cl−
and 1.0 mg mL−1 tween-80.


### 3.2. Effects of pH Value and Ionic Strength on RLS

The newly-formed AgCl (s) particles may be
ascribed to the higher electrostatic attraction between Ag+ and Cl−  than that of the
coexistent NO3− ion. Moreover, the RLS is relevant to the
size of the formed aggregated species. Hence, the pH value and ionic strength may exert certain influence on the attraction
strength and the dimension of suspension particles, and thus the RLS production
and its intensity. The RLS intensity of AgCl and
AgNO3 solution did not change with the variation of pH in range of
2.0–11.0. Sodium nitrate was used to maintain the ionic
strength of the solution. The unexpected observation is that both of the RLS intensity of AgCl and AgNO3 system hardly changed with the
concentration changes of added NaNO3 (figures are not presented).


### 3.3. Stability

The formation process of
aggregation particles generally includes three steps: nucleation, crystal
growth, and aggregation. The size of the particles is one important factor
affecting the RLS production and its intensity. To improve the reproducibility
of RLS intensity of a suspension system, it is crucial to impede the rapid
sedimentation of the particles. However, the AgCl
system is very stable in 40 minutes and the average deviation of RLS signal was
found to be lower than 3.80% (figure is not
presented). Moreover, the addition order of the reagents has little
effect on the RLS spectra.


### 3.4. Tolerance of Foreign Ions

There exist a number of cations and anions in drinks. To gain an insight into the RLS selectivity of
the AgCl for detection of Cl−  in drinks, the
interference of some commonly coexistent ions, such as Na+, Zn2+,
Ca2+, Fe2+, Mg2+, Al3+, CO32−,
SO32−, SO42−, PO43−,
K+, and I−, was investigated under the same conditions. The results demonstrated
that the CO32− and PO43− in excess of 1000 folds in
concentration relative to Cl− influenced the RLS of AgCl (s) particles.
This may be due to the formation of extended aggregate around AgCl particle cores by the relatively higher
negatively-charged ions of CO32− and PO43−.
I− in the same concentration relative to Cl−  strongly affected
the RLS signal. However, chloride is a prominent negatively charged ion of the
drinks. The concentration of I− is lower than
that of Cl−. Other ions
hardly interfered with the AgCl system (Table 1).
All concentration of other anions is lower than chloride ion. When the
concentration of each ion was the same as or somewhat lower than that of Cl−, no interference
was observed (results were not presented herein). Sodium ion (NaNO3)
hardly exerts interference as well as when its concentration reaches to ca. 10000 times that of Cl−. Therefore, the
tolerance levels of the interference of these coexisting ions in the samples
were very high and the assays can be performed without removing them.Table 1
Effect of interfering ions on the Cl− determination(a).
ion[ion]/[Cl−]Change in RLS (%)ion[ion]/[Cl−]Change in RLS (%)K+100000.75CO32−10002.01Ca2+100001.39PO43−10001.79Mg2+100001.56Ac−10000.88Zn2+100001.02HPO42−10001.09Fe2+100001.34H2PO4−10001.67Al3+100001.19I−16.32SO42−10002.34SO32−10001.86(a)The cationic ions were added in the form of nitrate, and the
anionic ions were added in the form of sodium. The
concentration of Cl−  is 8.52 ng mL−1.


## 3.1. Characteristics of the RLS Spectra

The RLS spectra of Cl−  and AgNO3 in water were shown in Figure 1(a) and (b), respectively. It can be seen that
the RLS intensity of AgNO3 solution is quite weak in the whole
scanning wavelength region. In contrast, upon addition of trace amount of Cl− to AgNO3 solution, a remarkably enhanced RLS with a maximum peak at 571 nm was observed
under the same conditions (1.0 mL of silver nitrate). The
addition of increasing Cl−  to the solution
leads to the gradual enhancement in RLS intensity, exhibiting a
concentration-dependent relationship. The production of RLS and its intensity are correlative with the formation of the aggregate and its
particle dimension in solution. Bearing on this point, we
inferred that the Cl− ion may displace
NO3− ion in AgNO3 (aq), forming a new AgCl (s) compound that
could be expected to be an aggregate. The size of AgCl (s)
particles may be much less than the incident wavelength, and thus the enhanced
light-scattering signal occurs under the given conditions. In this way, the resonance
light scattering formula could be applicable to the AgCl
system.Figure 1
RLS profile of
AgCl systems. (b)–(h) 6.99μg mL−1 of AgNO3,
1.0 mg mL−1 tween-80, and 0.0, 1.42, 2.84, 4.26, 5.68, 7.10,
8.52 ng mL−1 of Cl−. (a) 35.5 ng mL−1 of Cl−
and 1.0 mg mL−1 tween-80.


## 3.2. Effects of pH Value and Ionic Strength on RLS

The newly-formed AgCl (s) particles may be
ascribed to the higher electrostatic attraction between Ag+ and Cl−  than that of the
coexistent NO3− ion. Moreover, the RLS is relevant to the
size of the formed aggregated species. Hence, the pH value and ionic strength may exert certain influence on the attraction
strength and the dimension of suspension particles, and thus the RLS production
and its intensity. The RLS intensity of AgCl and
AgNO3 solution did not change with the variation of pH in range of
2.0–11.0. Sodium nitrate was used to maintain the ionic
strength of the solution. The unexpected observation is that both of the RLS intensity of AgCl and AgNO3 system hardly changed with the
concentration changes of added NaNO3 (figures are not presented).


## 3.3. Stability

The formation process of
aggregation particles generally includes three steps: nucleation, crystal
growth, and aggregation. The size of the particles is one important factor
affecting the RLS production and its intensity. To improve the reproducibility
of RLS intensity of a suspension system, it is crucial to impede the rapid
sedimentation of the particles. However, the AgCl
system is very stable in 40 minutes and the average deviation of RLS signal was
found to be lower than 3.80% (figure is not
presented). Moreover, the addition order of the reagents has little
effect on the RLS spectra.


## 3.4. Tolerance of Foreign Ions

There exist a number of cations and anions in drinks. To gain an insight into the RLS selectivity of
the AgCl for detection of Cl−  in drinks, the
interference of some commonly coexistent ions, such as Na+, Zn2+,
Ca2+, Fe2+, Mg2+, Al3+, CO32−,
SO32−, SO42−, PO43−,
K+, and I−, was investigated under the same conditions. The results demonstrated
that the CO32− and PO43− in excess of 1000 folds in
concentration relative to Cl− influenced the RLS of AgCl (s) particles.
This may be due to the formation of extended aggregate around AgCl particle cores by the relatively higher
negatively-charged ions of CO32− and PO43−.
I− in the same concentration relative to Cl−  strongly affected
the RLS signal. However, chloride is a prominent negatively charged ion of the
drinks. The concentration of I− is lower than
that of Cl−. Other ions
hardly interfered with the AgCl system (Table 1).
All concentration of other anions is lower than chloride ion. When the
concentration of each ion was the same as or somewhat lower than that of Cl−, no interference
was observed (results were not presented herein). Sodium ion (NaNO3)
hardly exerts interference as well as when its concentration reaches to ca. 10000 times that of Cl−. Therefore, the
tolerance levels of the interference of these coexisting ions in the samples
were very high and the assays can be performed without removing them.Table 1
Effect of interfering ions on the Cl− determination(a).
ion[ion]/[Cl−]Change in RLS (%)ion[ion]/[Cl−]Change in RLS (%)K+100000.75CO32−10002.01Ca2+100001.39PO43−10001.79Mg2+100001.56Ac−10000.88Zn2+100001.02HPO42−10001.09Fe2+100001.34H2PO4−10001.67Al3+100001.19I−16.32SO42−10002.34SO32−10001.86(a)The cationic ions were added in the form of nitrate, and the
anionic ions were added in the form of sodium. The
concentration of Cl−  is 8.52 ng mL−1.


## 4. Analytical Applications

### 4.1. Detection and Quantification Limits

The detection and quantification limits were calculated assb+3s and sb+10s,
respectively, where sb is the average RLS signal of ten blank solutions (16.99 μg mL−1 of silver nitrate) and s is the
standard deviation. When the RLS intensity at 571 nm was selected, the
detection and quantification limit were calculated to be 0.71 ng mL−1 and 1.42 ng mL−1, respectively,
indicating high sensitivity of this method for the determination of Cl−. The sensitivity of the RLS method is
prominently higher than that of turbidimetry
(results are not presented).


### 4.2. Calibration Curve and Assay of Samples

According to the above standard procedure, the calibration curve was obtained by plotting
the concentration of Cl− against the
intensity of RLS spectrum at 571 nm under the optimum conditions (Figure 2).
The linear regression equation using the least square method was ΔI=−26.97+27.25CCl− (ng mL−1)
(r2=0.9989) in the range of 1.42–8.52 ng mL−1 of Cl−. The proposed
method was applied to determine the concentration of Cl− in drinks and
the results were shown in Table 2.Table 2
Determination results of the chloride ion in drinks samples (n = 5).
SamplesRLS (μg mL−1)RSD (%)ISE (μg mL−1)RSD (%)Mineral waterNongfu spring20.351.3121.193.46LaoshanN.DN.DNestlé37.012.4337.951.54Robust37.381.5238.531.23Watsons36.912.0636.403.22Masterkong38.501.3339.081.68Pure waterWahahaN.DN.DNongfu springN.DN.DGreen tea65.791.0167.061.87Figure 2
The calibration curve of AgCl system at wavelength of 571 nm.


### 4.3. Recovery

To further examine the accuracy of the proposed RLS method, the recovery experiments were carried out
by adding known amounts of Cl− to drinks
diluted samples (Table 3). The
recovery results were obtained in the range of 94.08–105.63%,
which indicated that the proposed RLS method is practical to assay the Cl− in drinks.Table 3
Recovery test of some samples.
SampleCl−  in sample (μg)added Cl−  (μg)found (μg)recovery range (%)139.235.575.6–76.7102.53–105.63236.9135.570.3–71.994.08–98.58337.3835.571.9–73.897.18–102.53438.5035.572.2–75.994.92–105.35537.0135.570.5–74.194.37–104.51


### 4.4. Comparison of RLS and ISE Method

The proposed method was applied to determine the concentration of chloride ion in
drinks and the results were shown in Table2. To verify the veracity and
practicality, the samples were analyzed simultaneously by the currently used
ISE method (Table 2). It can be seen that the results determined by the RLS
method were almost in agreement with those by the ISE method. The average RSD
of the RLS method is 1.01%–2.43%, which is some lower than that of ISE method (1.23%–3.46%), which
proved that the RLS assay in drinks was practical.These two methods, RLS
and ISE, can give the similar results of chloride ion in drinks (Table2).
However, the operations of RLS and ISE methods were significantly different. As we know, ISE
method is an electrochemistry analytical method, in which the variation of
temperature, fluctuation of voltage, and treatment of electrode may bring on
significant influence on the determination though it features high selectivity.
However, the RLS method hardly suffers these factors.


## 4.1. Detection and Quantification Limits

The detection and quantification limits were calculated assb+3s and sb+10s,
respectively, where sb is the average RLS signal of ten blank solutions (16.99 μg mL−1 of silver nitrate) and s is the
standard deviation. When the RLS intensity at 571 nm was selected, the
detection and quantification limit were calculated to be 0.71 ng mL−1 and 1.42 ng mL−1, respectively,
indicating high sensitivity of this method for the determination of Cl−. The sensitivity of the RLS method is
prominently higher than that of turbidimetry
(results are not presented).


## 4.2. Calibration Curve and Assay of Samples

According to the above standard procedure, the calibration curve was obtained by plotting
the concentration of Cl− against the
intensity of RLS spectrum at 571 nm under the optimum conditions (Figure 2).
The linear regression equation using the least square method was ΔI=−26.97+27.25CCl− (ng mL−1)
(r2=0.9989) in the range of 1.42–8.52 ng mL−1 of Cl−. The proposed
method was applied to determine the concentration of Cl− in drinks and
the results were shown in Table 2.Table 2
Determination results of the chloride ion in drinks samples (n = 5).
SamplesRLS (μg mL−1)RSD (%)ISE (μg mL−1)RSD (%)Mineral waterNongfu spring20.351.3121.193.46LaoshanN.DN.DNestlé37.012.4337.951.54Robust37.381.5238.531.23Watsons36.912.0636.403.22Masterkong38.501.3339.081.68Pure waterWahahaN.DN.DNongfu springN.DN.DGreen tea65.791.0167.061.87Figure 2
The calibration curve of AgCl system at wavelength of 571 nm.


## 4.3. Recovery

To further examine the accuracy of the proposed RLS method, the recovery experiments were carried out
by adding known amounts of Cl− to drinks
diluted samples (Table 3). The
recovery results were obtained in the range of 94.08–105.63%,
which indicated that the proposed RLS method is practical to assay the Cl− in drinks.Table 3
Recovery test of some samples.
SampleCl−  in sample (μg)added Cl−  (μg)found (μg)recovery range (%)139.235.575.6–76.7102.53–105.63236.9135.570.3–71.994.08–98.58337.3835.571.9–73.897.18–102.53438.5035.572.2–75.994.92–105.35537.0135.570.5–74.194.37–104.51


## 4.4. Comparison of RLS and ISE Method

The proposed method was applied to determine the concentration of chloride ion in
drinks and the results were shown in Table2. To verify the veracity and
practicality, the samples were analyzed simultaneously by the currently used
ISE method (Table 2). It can be seen that the results determined by the RLS
method were almost in agreement with those by the ISE method. The average RSD
of the RLS method is 1.01%–2.43%, which is some lower than that of ISE method (1.23%–3.46%), which
proved that the RLS assay in drinks was practical.These two methods, RLS
and ISE, can give the similar results of chloride ion in drinks (Table2).
However, the operations of RLS and ISE methods were significantly different. As we know, ISE
method is an electrochemistry analytical method, in which the variation of
temperature, fluctuation of voltage, and treatment of electrode may bring on
significant influence on the determination though it features high selectivity.
However, the RLS method hardly suffers these factors.



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*Source: 745636-2008-05-19.xml*