DETERMINATION OF FLUORIDE IN WATER SAMPLES

A Laboratory Activity submitted as partial fulfillment of the requirements in Organic

Chemistry for Engineering and Applied Sciences Laboratory 2

John Carlo A. Delarmente

Jasper Vic R. Dela Cerna
King Zhytt C. Driz
Jasper M. Talsan
Learners

Maria Cecelia V. Almeda

Professor

May, 2022
 Activity 11
DETERMINATION OF FLUORIDE IN WATER SAMPLES

I. Introduction

As rainwater falls on the Earth and contacts soil, it dissolves minerals, which are washed into
streams and lakes. These waters, in turn, transport a variety of cations and anions to the oceans. Over
millions of years, this resulted in the high salt content of ocean water. Common cations include
sodium, potassium, calcium, and magnesium; common anions are chloride, sulfate, carbonate,
bicarbonate, and nitrate, although other cations and anions may be present, depending on the local
geologic media. Some ions are nutrients; others may be potentially toxic.
Fluoride is a natural element found in rocks and soil, and a low concentration of fluoride is
found in most waters. Fluoride prevents tooth decay by making teeth stronger and more resistant to
acid attacks. It also helps with slowing down or stopping the decay process.
When fluoride levels in water are at optimal levels, it helps to protect teeth against cavities. This
is why so many community water systems add additional fluoride – a process called “fluoridation.”
However, fluoride is harmful. Fluoride concentration of 5 mg/l can cause tooth decay more than the
case. And use of fluoride in drinking water at the 1.5 mg/l limit was put into the value.
This activity demonstrates the detection of fluoride ion in water samples, using Ion Selective
Electrode. This activity enables the students to be familiar with the entire method, from sample
preparation to detection. Note that electrodes measure activity, not concentration. In low–ionic
strength waters, the activity is essentially equal to concentration, but for higher ionic strengths,
important differences in these measurements are present.
The determination of electrolytes: sodium, potassium, and chloride are one of the most
important functions in a clinical laboratory. Electrolytes affect most metabolic processes. They serve
to maintain osmotic pressure, hydration of various body fluid compartments, proper body pH, and
regulation of heart and muscle functions. Ion-selective electrodes are used in clinical chemistry to
measure concentrations of ions in the blood, serum, plasma, cerebrospinal fluid, and urine.
An ion-selective electrode, also known as a specific ion electrode is a transducer or a sensor that
converts the activity of a specific ion dissolved in a solution into an electric potential. A pH-selective
glass electrode is immersed into a solution whose ion concentration is to be measured. When the ion-
selective electrode is immersed in a solution, the solution potential difference develops between the
inner and the outer layers of the membrane of the electrode which is related to the ionic
concentration of the solution. According to the Nernst equation, the voltage is theoretically on the
logarithm of the ionic activity. Most analyzers have a common part such as a removable reagent
pack, display screen, keyboard, sample probe, sensors or electrodes, peristaltic pump, tubing, and
reagent pack or module.
The analyzer requires daily maintenance of cleaning the fluid path to remove protein deposits.
Cleaning is performed daily at the beginning of the day before the samples are run. After cleaning,
the instrument performs an automatic calibration. The instrument is set for two-point auto-
calibration every 8 hours. The calibration performed by the machine is assessed by obtaining
calibration values for each sensor. The values should be within the acceptable ranges. The values are
to be checked daily and should be documented. Acceptable ranges are displayed as bench-aid near
the equipment. If the acceptance criteria are breached, then perform maintenance as per the
manufacturer’s instructions. A few causes of calibration are sensor or electrode failure, reagent
expiry, and air bubbles in the probe. After calibration and before patient sample analysis, quality
control is analyzed to establish the equipment’s performance. The performance of the analyzer must
be monitored using quality controls. Each lot of analyzer control material contains an insert sheet, an
indication of the expected ranges for each analyte. The analyzer stores quality control results for
three levels, a maximum of 31 for each level. Results for each analyte are compared to the quality
reference control limits.
All standard precautions are need to be followed while handling samples. Serum or plasma (free
from hemolysis) is the recommended specimen. Separate serum from red blood cells as soon as
possible.
Pre-analytical errors can occur due to hemolysis, storage, time, and temperature. Other factors
includes skeletal muscle activity, special care taken to loosen the tourniquet before sample draw,
patient should not clench and unclench the fist repeatedly, use fresh sample for analysis, store the
sample in a stopper tube if the analysis is delayed.
Samples containing particulate matter are centrifuged and material is removed before analysis.
Grossly lipemic specimens should be cleared by ultracentrifugation. Pseudo hyponatremia maybe
seen lipemic specimens as a result of fluid displacement. Separate serum immediately after
centrifugation. Potassium from the red cells diffusing into the serum, gives falsely elevated results.
Hemolysis shows no significant interference with sodium or chloride. Turbid urine samples need to
cleared through centrifugation. In very rare cases, gammopathy type IgM causes unreliable results.
Ion-selective electrodes are not based on reduction-oxidation processes but on selective binding
of one type of ion to a membrane, which generates a potential. The combination of glass pH
electrode contains a complete galvanic cell which two electrodes in a single probe. As the pH of the
external solution changes, the electric potential across the glass membrane changes.
 II. Method

Fluoride has many sources, it can naturally be found in solid and water sources; it can even be
found in foods. It is also produced synthetically used in drinking water, toothpaste, mouthwashes, and
other chemically-iduced products. In this activity, fluoride concentration in water sample is to be
determined by Ion-Selective Electrode (ISE) method.

The following were the apparatus and the solutions used in this experiment:

Apparatus

● ion-selective electrode meter

● fluoride ion-selective electrode
● magnetic stirrer with TFE-coated stirring bracket
● beaker
● pipette

Solutions

● standard fluoride solution

● fluoride buffer (TISAB - Total IOnic Strength Adjusting Buffer) solution
● distilled water
● water sample

Preparation of Fluoride Standards

1. Prepare three standards by diluting 2.5, 5.0, and 10ml of standard fluoride solution to 100ml with
distilled water.
2. Take 25ml of the sample solution in a 100ml - beaker.
3. Add 25ml of total-ionic strength adjustment buffer (TISAB) to the same beaker in (2).

Calibration of Ion Selective Electrode Meter (ISEM)

1. Clean the electrode of theISEM with distilled water and dry it using tissue paper.
2. Place the beaker with the 2.5ml fluoride solution on a stirring plate and add the magnetic stirring
bar. After which, dip the Fluoride ISE in the solution.
3. Turn on the stirring plate; when the reading becomes stable, turn it off to get the accurate
measurement.
4. Then again, clean the electrode using distilled water and dry it using a tissue paper.
5. Repeat steps (2) and (1) but this time with the 5ml fluoride solution.
 6. Repeat steps (2) and (1) but this time with the 10ml fluoride solution.

Determination of Fluoride Concentration in Water Sample

1. Clean the electrode of the ISEM with distilled water and dry it using tissue paper.
2. Place the beaker with the water sample on a stirring plate and add the magnetic stirring bar. After
which, dip the Fluoride ISE in the solution.
3. Turn on the stirring plate; when the reading becomes stable, turn it off to get the accurate
measurement.
4. Then again, clean the electrode using distilled water and dry it using a tissue paper.

III. Procedure

Using an Ion Selective Electrode

- Wear proper protective equipment

- Put the meter to standby mode
- Immerse the electrodes into the solution
- Rinse the electrode over a large waste beaker using squirt bottle
- Rinse the reference electrode over a large waste beaker using squirt bottle
- Rinse the stir bar
- Rinse the beaker three times
- Pour out the rinses into a waste beaker
- Change the readout on the meter to the millivolt setting by pressing the absolute button.
- Pour the solution into the rinsed sample beaker
- Stir the solution using the rinsed stir bar and slowly increase the stir rate
- Wait for the solution to stabilize
- Record the potential of the sample and remove the stir bar using a magnetic wand
- Rinse the sample beaker for the next process
- Fill the beaker with a standard fit for the sample being analyzed
- Design a calibration curve.

ISE Calibration - Preparation of Standards and Samples

- a
 IV. Results and Discussion

Sample Concentration of Fluoride (mg/L)

1 2.5

2 5.0

3 10.0

Unknown Water 0.11

Table 1. Fluoride Concentration in an Unknown Water Sample.

The table above shows that the water sample given has a fluoride level of 0.11 mg/L, so it
is suitable for drinking. Since the acceptable limit of fluoride in drinking water is less than 1
mg/L.

V. References:

Dunnivant, F. M. , Environmental Laboratory Exercises for Instrumental Analysis and Environmental

Chemistry, John Wiley & Sons, Inc., Hoboken, New Jersey, 2004.

IS 3025 (Part 60) -2008 : Method of Sampling and Test (Physical and Chemical) for Water and
Wastewater : Fluoride , First Revision.

Griffin SO, Regnier E, Griffin PM, Huntley VN. Effectiveness of fluoride in preventing caries in adults. J
Dent Res. 2007;86(5):410–414.

Arnold FJ, Dean HT. Effect of fluoridated public water supply on dental caries prevalence. Public Health
Rep. 1956;71:652–658.

ELECTROLYTES BY ISE METHOD: https://www.youtube.com/watch?v=XlbXXAH8RdQ

Ion-Selective Electrodes: https://www.youtube.com/watch?v=44h-z4b_IME
Using an Ion Selective Electrode: https://www.youtube.com/watch?v=NHByG4g5-RE
ISE Calibration - Preparation of Standards and Samples - Part 1: https://www.youtube.com/watch?
v=hwy6Frl0jX4
ISE Calibration - Part 2: https://www.youtube.com/watch?v=WR27nliMDzk
Ion Selective Electrodes - Tech Tips with Vernier: https://youtu.be/sgXm9vuO8Zk
Determination of Fluoride by ISE - Video Tutorial: https://www.youtube.com/watch?
v=Zr7haKMlwqs
Introduction: Fluoride Determination by Ion-Selective Electrode: https://www.youtube.com/watch?
v=_JijCxatEW4

Chemistry - Ion Selective Electrode experiment: https://www.youtube.com/watch?

v=XMJtdBzsWVY

Determination of Fluoride in Water virtual lab:

https://ee2-nitk.vlabs.ac.in/exp/flouride-in-water/simulation.html