o Procedures

I. Standard Water Quality Tests

• Alkalinity Test
• Chloride Test
• Total Hardness Test
• Calcium Hardness Test
• Magnesium Hardness Test
• Nitrate Test

II. Spectrophotometric Test Procedures

• How to Use a Spectrophotometer (Instructional Step)
• Turbidity Test
• Total Chlorine Test
• Free Chlorine Test – Method 1
• Free Chlorine Test – Method 2
• Phosphate Test
• Eliminox Test
• Zinc Test
• High-Level Iron Test
• Low-Level Iron Test
• Silica Test
• Sulphate Test
 Precautions before water analysis:
🔬 Precautions Before Water Analysis:
🧼 Sample Collection & Handling

1. ✅ Use Clean & Sterile Containers

– Prevents contamination during sampling.
2. ✅ Label Samples Clearly
– Include date, time, location, and type of analysis.
3. ✅ Use Appropriate Sample Bottles
– Glass for organics, plastic for inorganics.
4. ✅ Avoid Exposure to Sunlight & Heat
– Some analytes degrade quickly under these conditions.
5. ✅ Store and Transport Samples Properly
– Use coolers or refrigeration to maintain sample quality.

⚗️ Lab Preparation & Equipment

6. ✅ Calibrate Instruments Before Use

– Ensures accurate readings for pH, DO, etc.
7. ✅ Check Expiry Dates of Reagents
– Expired chemicals = incorrect results.
8. ✅ Use Deionized or Distilled Water
– For rinsing and reagent prep, to avoid contamination.
9. ✅ Perform a Blank Test (if required)
– Helps detect contamination in tools or reagents.
10. ✅ Avoid Soap Residue in Glassware
– Soap can interfere with chemical analysis.

🧤 Safety & Personal Hygiene

11. ✅ Wear Proper PPE

– Gloves, lab coat, goggles – always!
12. ✅ Avoid Touching Inner Surfaces
– Especially bottle caps and necks – prevent contamination.
13. ✅ Dispose of Waste Properly
– Follow chemical disposal guidelines.
14. ✅ Ensure Proper Ventilation
– Especially when working with volatile or hazardous chemicals.
📋 Testing Procedures & Accuracy

15. ✅ Follow Standard Operating Procedures (SOPs)

– Consistency is key to reliable data.
16. ✅ Avoid Over-Dilution or Incorrect Prep
– Always measure accurately.
17. ✅ Minimize Time Between Collection & Testing
– For more reliable results, especially biological.
18. ✅ Know Holding Times for Each Test
– Microbial samples, for example, must be tested within hours.
19. ✅ Avoid Air Bubbles (BOD/DO Tests)
– Air can falsely raise oxygen levels.
20. ✅ Document Every Step & Observation
– Good lab records = traceable and validated data.

🎯 Tip:
Clean workspace = clear results. Always start and finish with a tidy station!
 STANDARD WATER QUALITY TESTS

#1: Alkalinity Test Procedure

Apparatus:

• Flask
• Measuring Cylinder
• Burette

Reagents:

• Methyl Orange (M.O.) Indicator

• Hydrochloric Acid (HCl) N/50

Procedure:

1. Take a 20 mL water sample using a measuring cylinder. Pour the sample into a clean flask
(as shown in figure 1).
2. Add 2 to 3 drops of Methyl Orange (M.O.) indicator into the water sample. Gently swirl
the flask to mix the indicator with the water.
o Result: The solution should turn yellow if the sample contains alkaline substances
(e.g., bicarbonates, carbonates) (as shown in figure 2).
Figure 1
3. Titrate with Hydrochloric Acid (HCl) N/50:
o Using a burette, slowly add HCl N/50 drop by drop into the water sample while
continuously swirling the flask.
o Continue adding the acid until the color of the solution changes from yellow to red
(as shown in figure 3).
▪ The yellow color indicates an alkaline solution, and the red color indicates
the endpoint, where the alkalinity has been neutralized by the acid.
Figure 2
4. Record the burette reading:
o Note the volume of acid (in mL) used to reach the red color. This is the burette
reading.
o Calculate the alkalinity using the following formula:
Alkalinity (ppm)=Burette Reading (mL)×50
o The result is the alkalinity of the water sample, expressed in parts per million (ppm).
Figure 3
Important Notes:

• Methyl Orange (M.O.) Indicator: The color change from yellow to red is the key indicator of
the test's endpoint. Ensure that the color change is distinct and clear.
• Accuracy: Always ensure you add the acid slowly to avoid overshooting the endpoint. Stir
continuously to ensure even mixing of the sample.
• Precision: Record the burette reading carefully, and if needed, repeat the test for better
accuracy.
#2: Chloride Test Procedure
Apparatus:

• Flask
• Measuring Cylinder
• Burette

Reagents:

• Ferric (Fe) Indicator

• Silver Nitrate (AgNO₃) N/10
• Ammonium Thiocyanate (NH₄SCN) N/10

Procedure:

1. Prepare the Water Sample:

o Take a 50 mL water sample using a clean measuring cylinder.
o Pour the sample into a clean flask (as shown in figure 1).
2. Add Ferric Indicator:
o Add 20 mL of Ferric (Fe) Indicator to the sample.
o Gently swirl the flask to mix. The solution will take on a yellowish color (as shown in Figure 1
figure 2).
3. Add Ammonium Thiocyanate (NH₄SCN):
o Add 2 to 3 drops of Ammonium Thiocyanate (NH₄SCN) to the solution.
o Gently mix the solution. The sample should now turn a brownish color (as shown in
figure 3). due to the complex formed between the ferric ions and thiocyanate ions. Figure 2
4. Titrate with Silver Nitrate (AgNO₃): 2
o Using a burette, slowly add Silver Nitrate (AgNO₃) N/10 drop by drop into the
sample while continuously swirling the flask.
o Continue titrating until a white precipitate forms in the solution (as shown in
figure 4). This indicates that all chloride ions in the sample have reacted with the Figure 3
silver ions.
5. Add Excess Silver Nitrate:
o Add 3 mL of excess Silver Nitrate (AgNO₃) to ensure that all chloride ions have
reacted and there is sufficient AgNO₃ present for the next titration. 3
6. Titrate with Ammonium Thiocyanate (NH₄SCN):
o Now, titrate the excess Silver Nitrate (AgNO₃) with Ammonium Thiocyanate
Figure 4
(NH₄SCN) from the burette.
o Slowly add NH₄SCN drop by drop while swirling the flask, until the solution turns a
light brownish color (as shown in figure 5). This indicates the endpoint, where all
excess silver ions have been reacted with the thiocyanate ions.
7. Record the Burette Readings: Figure 5
o Note the burette readings for both AgNO₃ and NH₄SCN.
o Subtract the NH₄SCN burette reading from the AgNO₃ burette reading to determine
the volume of NH₄SCN used to titrate the excess AgNO₃.
8. Calculate Chloride Concentration:
 o The difference between the burette readings represents the amount of NH₄SCN used
to neutralize the excess AgNO₃.
o Calculate the chloride concentration in the sample using the following formula:

Chloride (ppm) = (AgNO₃ burette reading−NH₄SCN burette reading) × 117

▪ The result will give you the chloride concentration in parts per million (ppm).

Key Notes:

• White Precipitate Formation: The appearance of a white precipitate during the AgNO₃
titration indicates that chloride ions have reacted with silver nitrate.
• Light Brownish Color: The light brownish color after titrating with NH₄SCN indicates that all
excess silver ions have reacted, marking the endpoint of the second titration.
• Excess AgNO₃: Adding excess AgNO₃ ensures that there is enough silver nitrate to react with
all chloride ions in the sample, providing an accurate reading in the second titration with
NH₄SCN.
#3: Total Hardness Procedure

Apparatus:

• China Dish
• Stirrer
• Measuring Cylinder
• Burette

Reagents:

• Total Hardness (T.H.) Indicator

• Ammonia Buffer
• EDTA N/50

Procedure:

1. Prepare the Solution:

o Take distilled water in a clean china dish (approximately 50 mL) (as shown
in figure1).
o Add ammonia buffer to the distilled water in the china dish. This helps to
maintain the pH of the solution at the proper level for the titration process.
2. Add Total Hardness Indicator:
o Add a few drops of Total Hardness (T.H.) Indicator to the solution. Stir gently. Figure 1
3. Add EDTA Solution:
o Slowly add EDTA N/50 solution to the mixture while stirring gently. The
solution will change to light blue (as shown in figure 2), which indicates that
the EDTA is binding with the calcium and magnesium ions (which are
responsible for hardness).
4. Add the Water Sample:
o Add 10 mL of the water sample to the solution in the china dish. The solution
will turn pink (as shown in figure 3), indicating the presence of hardness ions Figure 2
(calcium and magnesium) in the water sample.
5. Titrate with EDTA:
o Using the EDTA N/50 solution in the burette, slowly titrate the pink solution
drop by drop, while continuously stirring. Titrate until the pink color
changes to light blue (as shown in figure 2), This indicates that all the
hardness ions have reacted with the EDTA, and the titration is complete.
Figure 3
6. Calculate the Total Hardness:
o Record the burette reading (the volume of EDTA solution used).
o Multiply the burette reading (in mL) by 100 to calculate the total hardness of
the water sample in ppm (parts per million).

Total Hardness (ppm)=Burette Reading (mL)×100

Key Notes:

• Yellow Color: The initial yellow color indicates that calcium and magnesium ions are
present in the sample, contributing to hardness.
• Pink Color: The solution will turn pink when the water sample is added, showing that
hardness ions are present.
• Light Blue Color: The change from pink to light blue signals the endpoint of the
titration, meaning all hardness ions have reacted with EDTA.
• Add EDTA drop by drop as you approach the endpoint to avoid overshooting and
ensure an accurate result.

#4 Calcium Hardness Procedure

Apparatus:

• China Dish
• Stirrer
• Measuring Cylinder
• Burette

Reagents:

• Calcium Hardness (C.H.) Indicator

• Sodium Hydroxide (NaOH) N/1
• EDTA N/50

Procedure:

1. Prepare the Solution:

o Take distilled water in a clean china dish (approximately 50 mL).
o Add 30 mL of Sodium Hydroxide (NaOH) N/1 to the distilled water (as shown
in figure 1). This increases the pH of the solution and makes it alkaline, which
is necessary for the calcium ions to react with the EDTA during the titration.
2. Add Calcium Hardness Indicator: Figure 1
o Add 4mg/L to 5mg/L drops of Calcium Hardness (C.H.) Indicator to the
solution. Stir gently.
o The solution will turn Purple as the indicator binds with calcium ions present
in the solution (as shown in figure 2).
3. Add Raw Water Sample:
o Add 10 mL of the raw water sample to the solution. The solution will turn
pink, indicating that calcium ions are present in the water sample as in Figure 2
figure 3.
 4. Titrate with EDTA (First Titration):
o Fill the burette with EDTA N/50 solution.
o Titrate drop by drop, stirring gently after each addition, until the solution
changes from pink to purple (as shown in figure 2).
o The appearance of the purple color indicates that all the calcium ions in the
sample have reacted with the EDTA. Figure 3
o Zero the burette before starting to ensure accurate readings.
5. Add the Water Sample:
o Add 10 mL of the water sample to the solution.
o The solution will turn pink again, and you will need to titrate again with EDTA.
6. Titrate with EDTA (Second Titration):
o Titrate with EDTA N/50 solution again, drop by drop, until the solution
changes from pink to purple.
o This final purple color indicates that the titration is complete, and all the
calcium ions have reacted with the EDTA.
7. Calculate Calcium Hardness:
o Record the burette reading (the volume of EDTA solution used in the
titration).
o Multiply the burette reading (in mL) by 100 to calculate the calcium hardness
in ppm (parts per million).

Calcium Hardness (ppm)=Burette Reading (mL)×100

Key Notes:

• Blueish Color: The appearance of a blueish color after titration indicates that all
calcium ions have reacted with EDTA.
• Pink Color: The solution will turn pink when calcium ions are present in the water
sample.
• Titration Endpoint: The endpoint of the titration is indicated by a color change from
pink to blueish. This indicates that all calcium ions have been neutralized.

#5 Magnesium Hardness Procedure

Magnesium Hardness = Total hardness - Calcium Hardness

 Spectrophotometer

📘 Introduction
A spectrophotometer is a commonly used analytical instrument in scientific laboratories that measures
the amount of light absorbed by a solution. It operates by passing a beam of light through a sample and
detecting how much of that light is absorbed at a specific wavelength. This technique is widely used to
determine the concentration of substances in solution and plays a vital role in various fields like biology,
chemistry, environmental science, pharmaceuticals, and clinical diagnostics.

🔹 Hach DR 3900 Spectrophotometer Overview

The Hach DR 3900 is a high-performance benchtop visible spectrum spectrophotometer designed for
precise water quality analysis. It offers a wavelength range of 320–1100 nm and supports over 240 pre-
programmed test methods, making it suitable for various applications such as municipal water, surface
water, and QA/QC laboratories.

📘 Picture:
🔧 Key Features

• Optical System: Spectral reference beam technology

• Light Source: Gas-filled tungsten lamp
• Wavelength Range: 320–1100 nm
• Spectral Bandwidth: 5 nm ± 1 nm
• Wavelength Accuracy: ±1.5 nm (340–900 nm)
• Photometric Accuracy: ±5 Abs at 0.0–0.5 Abs
• Sample Compatibility: 13 mm round and 10/50 mm rectangular cuvettes
• Display: 7" TFT color touchscreen
• Connectivity: USB, Ethernet, optional RFID module
• Power Supply: 110–240 V AC, 50/60 Hz
• Dimensions: 350 x 255 x 151 mm
• Weight: 4.2 kg
• Operating Conditions: 10–40°C, <80% relative humidity
• Storage Conditions: -30–60°C

🔹 How to use DR 3900 Spectrophotometer

✅ 1. Power On the Instrument

• Plug in the DR3900 and press the power button.

• Wait a few minutes for it to fully start up and warm the light source

✅ 2. Choose the Test Method

• On the touchscreen, either:

o Select your test from the menu (e.g., COD, nitrate, phosphate),
o Or scan the barcode on the reagent pack if available.

💡 The DR3900 automatically loads the correct wavelength and settings when you scan Hach reagents.

✅ 3. Prepare the Blank and Sample

• Use 10 mL square cell for both the blank and the sample.
• Follow the reagent instructions carefully:
o Fill one cuvette with the blank (usually DI water + reagents, no sample).
o Fill another cuvette with your prepared sample.

📌 Make sure both are clean and wiped with a tissue. Handle square cell by the top to avoid
fingerprints.

✅ 4. Zero the Instrument (Blank Calibration)

• Place the blank cell in the sample chamber.

• Close the lid securely.
 • Tap “Zero” on the touchscreen.
• Wait for the display to show 0.000 (absorbance) or say “Zero completed.”

✅ 5. Run the Test

• Take out the blank and insert your sample cell.

• Close the lid again.
• Tap “Read” or “Start” to begin measurement.

⏱️ In a few seconds, the DR3900 will show your result in mg/L, ppm, or Abs depending on the
method.

✅ 6. Record or Save the Results

• Write down the reading in your lab notebook

✅ 7. Clean Up and Repeat (if needed)

• If testing more samples, clean and refill the 10 mL sample cell and repeat steps 5–6.
• Always rinse cuvettes between different samples.

✅ 8. Shutdown After Use

• When finished, turn off the instrument.

• Clean and store the 10 mL cuvettes properly.
• Cover the spectrophotometer to protect it from dust.

📌 Notes:
• Only use clean 10 mL square cell – scratches, bubbles, or dirt will affect results.
• Make sure to align the cell correctly in the holder (some have alignment marks).
• Always zero with a blank before starting a new test or changing parameters.
 Spectrophotometric Test Procedures

🔹 Phosphate Test Procedure

APPARATUS:

• 10 mL Square cell

REAGENTS:

• Phosphate Reagent (Molybdovanadate Dropper)

PROCEDURE:

1. Prepare the Sample:

o Take 10 mL of the water sample and transfer it into a clean 10 mL square cell
as shown in figure 1.
2. Select the Test Program:
o On the DR3900 spectrophotometer, set the program code to 480 (Phosphate
test).
3. Add the Reagent:
o Add 8 drops of the phosphate reagent (Molybdovanadate) to the cell. Figure 1
o Close the cuvette and gently shake to mix the solution.
4. Wait for Color Development:
o The sample will develop a yellowish color as shown in figure 2, indicating the
presence of phosphate.
5. Set the Timer:
o Set the timer for 7 minutes to allow the color to fully develop.
6. Zero the Instrument: Figure 2
o Insert a blank cell (distilled water) into the spectrophotometer.
o Press “Zero” to calibrate the instrument.
7. Read the Sample:
o After the 7-minute timer is complete, place the cell with the sample into the
instrument.
o Press “Read” to measure the absorbance.
8. Record the Result:
o The spectrophotometer will display the phosphate concentration in ppm (parts
per million).
🔹 Turbidity Test Procedure

APPARATUS:

• 10 mL square cell

REAGENTS:

• No reagent required for turbidity testing.

PROCEDURE:

1. Prepare the Sample:

o Take 10 mL of the water sample and pour it into a clean 10 mL cell.
2. Select the Test Program:
o On the DR3900 spectrophotometer, set the program code to 265 (Turbidity
test).
3. Zero the Instrument:
o Fill a separate 10 mL cell with distilled water to use as a blank.
o Insert the blank into the instrument and press “Zero” to calibrate.
4. Measure the Sample:
o Remove the blank and insert the sample cell.
o Close the lid and press “Read” to take the measurement.
5. Record the Result:
o The DR3900 will display the turbidity value in FAU (Formazin Attenuation
Units).

🔹 Sulphate Test Procedure

APPARATUS:

• 10 mL Square cell

REAGENTS:

• Sulphate Reagent Pillow (Vario Sulfa)

PROCEDURE:
 1. Prepare the Sample:
o Pour 10 mL of the water sample into a clean 10 mL cell as shown in figure 1.
2. Select the Test Program:
o On the DR3900 spectrophotometer, set program code 680 (Sulphate Test).
3. Zero the Instrument:
o Fill another cell with 10 mL of distilled water.
o Insert it into the instrument and press “Zero” to calibrate.
4. Add Reagent:
o Open one Vario Sulfa reagent pillow.
Figure 1
o Add the entire contents into the sample cell.
o Cap and shake gently until the powder dissolves completely as shown in figure
2.
5. Set the Timer:
o Allow the reaction to develop for 5 minutes.
6. Measure the Sample:
o After the timer ends, insert the sample cell into the instrument.
o Close the lid and press “Read” to take the measurement.
Figure 2
7. Record the Result:
o The DR3900 will display the sulphate concentration in ppm (mg/L).

⚠️ Note (For High Sulphate Concentrations):

If the sulphate concentration is too high and out of range:

1. Dilute the Sample:

o Mix 2 mL of the sample with 48 mL of distilled water in a clean container as shown
in figure 3.
o Mix thoroughly.
2. Re-Test:
o Take 10 mL from the diluted solution into a cell.
o Repeat the procedure from Step 2 using this diluted sample.
3. Multiply the Result: Figure 3
o After measurement, multiply the result by 25 to get the actual concentration.
🔹 Total Chlorine Test Procedure

APPARATUS:

• 10 mL Square cell

REAGENTS:

• Chlorine Powder Pillow (DPD Total Chlorine Reagent)

PROCEDURE:

1. Prepare the Sample:

o Pour 10 mL of the water sample into a clean 10 mL cell.
2. Select the Test Program:
o On the DR3900 spectrophotometer, set program code 80 (Total Chlorine Test).
3. Add Reagent:
o Open one Total Chlorine reagent pillow.
o Add the full contents into the sample cell. Figure 1
o Cap and shake gently until the powder fully dissolves then pink color appears
shown in figure 2.
4. Set the Timer:
o Start a timer for 3 minutes to allow the reaction to complete.
5. Zero the Instrument:
o Fill another cell with 10 mL of distilled water.
o Insert the blank into the spectrophotometer and press “Zero” to calibrate.
6. Measure the Sample:
o After the 3-minute reaction time, insert the sample cell into the instrument.
Figure 2
o Press “Read” to begin measurement.
7. Record the Result:
o The DR3900 will display the total chlorine concentration in ppm (mg/L).

🔹 Free Chlorine Test Procedure

APPARATUS:

• 10 mL Square cell

REAGENTS:

• DPD Free Chlorine Powder Pillow

PROCEDURE:

1. Prepare the Sample:

o Pour 10 mL of the water sample into a clean 10 mL cell.
2. Select the Test Program:
o On the DR3900 spectrophotometer, set program code 80 (Free Chlorine Test).
3. Add the Reagent:
o Open one DPD Free Chlorine reagent pillow.
o Add the entire contents to the sample cell.
Figure 1
o Cap and shake gently to dissolve the powder.
4. Observe Color Change:
o A pink color will develop if free chlorine is present in the sample as shown in
figure 2.
5. Set the Timer:
o Start a 3-minute timer to allow for complete color development.
6. Zero the Instrument:
o Fill another cell with 10 mL of distilled water.
o Insert the blank into the instrument and press “Zero” to calibrate.
7. Measure the Sample: Figure 2
o After the timer ends, insert the sample cell into the DR3900.
o Press “Read” to measure the chlorine concentration.
8. Record the Result:
o The DR3900 will display the free chlorine concentration in ppm (mg/L).

🔹 Free Chlorine Test Procedure (Orthotolidine Method)

APPARATUS:

• 10 mL Square cell

REAGENTS:

• Chlorine Dropper (Orthotolidine Solution)

PROCEDURE:

1. Prepare the Sample:

o Pour 10 mL of the water sample into a clean 10 mL cell as shown in figure 1.
2. Select the Test Program:

Figure 1
 o On the DR3900 spectrophotometer, set program code 9006 (Free Chlorine –
Orthotolidine method).
3. Add Reagent:
o Add 6 drops of Orthotolidine (chlorine dropper) to the sample.
o Cap the cuvette and shake gently to mix.
4. Observe Color Change:
o The solution will turn a yellow-greenish color if free chlorine is present as
shown in figure 2.
5. Zero the Instrument:
o Fill another cuvette with 10 mL of distilled water.
o Insert the blank into the instrument and press “Zero”.
Figure 2
6. Measure the Sample:
o Immediately insert the sample cell into the spectrophotometer.
o Press “Read” to measure the chlorine concentration.
7. Record the Result:
o The DR3900 will display the free chlorine concentration in ppm (mg/L).

🔹 Zinc Test Procedure

APPARATUS:

• 10 mL Square cell
• 50 mL Beaker

REAGENTS:

• Zinc Reagent Pillow (Zinc Ove)

• Zinc Dropper (Cyclohexanone)

PROCEDURE:

1. Prepare the Sample:

o Take 20 mL of the water sample in a clean 50 mL beaker.
2. Add the Reagent Pillow:
o Add one Zinc reagent pillow (Zinc Ove) to the sample.
o Shake well for 30 seconds until a red color develops as shown in figure 1.
3. Select the Test Program:
o On the DR3900 spectrophotometer, set program code 780 (Zinc Test).
4. Transfer to Cell:
o Pour 10 mL from the beaker into a clean 10 mL cell.
Figure 1
 5. Zero the Instrument:
o Insert this cell into the spectrophotometer and press “Zero” to calibrate.
o Discard this cell after zeroing.
6. Prepare the Final Sample:
o Pour a fresh 10 mL of the treated sample into another clean cell.
o Add 1 mL of Zinc dropper (Cyclohexanone) and mix gently.
o A darker red color indicates the presence of zinc as shown in figure 2.
7. Set the Timer:
o Allow the reaction to develop for 3 minutes.
8. Read the Sample:
o After the timer ends, insert the cell into the spectrophotometer.
o Press “Read” to measure the zinc concentration.
9. Record the Result: Figure 2
o The instrument will display the zinc value in ppm (mg/L).

🔹 Silica Test Procedure

APPARATUS:

• 10 mL Square cell

REAGENTS:

• Silica 1 Dropper (Molybdate)

• Silica 2 Pillow (Citric Acid)
• Silica 3 Pillow (Amino Acid)

PROCEDURE:

1. Prepare the Sample:

o Pour 10 mL of the water sample into a clean 10 mL cell as shown in figure 1.
2. Select the Test Program:
o On the DR3900 spectrophotometer, set program code 651 (Silica Test).
3. Add Silica 1 Reagent:
o Add 14 drops of Silica 1 (Molybdate dropper) into the cell as shown in figure
2.
o Mix gently and start the first timer (follow instrument instructions for timing, Figure 1
typically ~2 minutes).
4. Add Silica 2 Reagent:
o After the first timer finishes, add one Silica 2 reagent pillow (Citric Acid).
o Shake or mix gently, then start the second timer (typically ~2 minutes).
5. Zero the Instrument:

Figure 2
 oAfter the second timer ends, insert the cell (with sample and both reagents) into
the instrument.
o Press “Zero” to calibrate.
6. Add Silica 3 Reagent:
o Remove the cell, add one Silica 3 reagent pillow (Amino Acid).
o Shake gently until a blue color develops as shown in figure3.
o Start the third timer (typically ~2 minutes) for full color development.
7. Read the Sample:
o After the timer finishes, insert the cell into the spectrophotometer.
o Press “Read” to measure the result.
8. Record the Result: Figure 3
o The DR3900 will display the silica concentration in ppm (mg/L).

⚠️ Note (For High Silica Concentrations):

If the silica concentration is too high (above instrument range):

1. Dilute the Sample:

o Mix 2 mL of the sample with 48 mL of distilled water in a clean container (total 50
mL) as shown in figure 4.
o Mix thoroughly.
2. Re-Test:
o Take 10 mL from the diluted solution into a cell.
o Repeat the same procedure from Step 2.
3. Multiply the Result:
o After measurement, multiply the result by 25 to get the actual silica concentration.
Figure 4

🔹 Eliminox Test Procedure

APPARATUS:

• 10 mL Square cell
• 50 mL Beaker

REAGENTS:

• Eliminox Reagent Pillow (DEHA 1)

• Eliminox Dropper (DEHA 2)
PROCEDURE:

1. Prepare the Sample:

o Take 25 mL of the water sample in a clean 50 mL beaker as shown in figure 1.
2. Add Reagents:
o Add one Eliminox reagent pillow (DEHA 1) to the beaker.
o Then add 0.5 mL of Eliminox dropper (DEHA 2).
o Stir or swirl gently to mix.
3. Incubate in Dark:
o Place the beaker in a dark area and allow the reaction to develop for 10
minutes.
o A purple color will appear if Eliminox (DEHA) is present as shown in figure 2. Figure 1
4. Transfer Sample to Cuvette:
o After 10 minutes, pour 10 mL of the reacted solution into a clean 10 mL cell.
5. Zero the Instrument:
o Insert a cell filled with 10 mL of distilled water into the spectrophotometer.
o Press “Zero” to calibrate.
6. Measure the Sample:
o Remove the blank and insert the sample cell.
o Press “Read” to begin measurement. Figure 2
7. Record the Result:
o The DR3900 will display the Eliminox (DEHA) concentration.

🔹 High-Level Iron (HLI) Test Procedure

APPARATUS:

• 10 mL Square cell

REAGENTS:

• Iron Reagent Pillow (Vario Ferro)

PROCEDURE:

1. Prepare the Sample:

o Pour 10 mL of the water sample into a clean 10 mL cell as shown in figure 1.
2. Add Reagent:
o Add one Iron reagent pillow (Vario Ferro) to the cuvette.
o Cap and shake gently to dissolve the reagent completely it shows light brown
color if iron is present in it as shown in figure 2.
Figure 1
 3. Select the Test Program:
o On the DR3900 spectrophotometer, set program code 265.
4. Start the Reaction Timer:
o Set a 3-minute timer to allow full color development.
5. Zero the Instrument:
o Fill another cell with 10 mL of distilled water. Figure 2
o Insert it into the spectrophotometer and press “Zero” to calibrate.
6. Read the Sample:
o After the timer ends, insert the sample cell into the instrument.
o Press “Read” to take the measurement.
7. Record the Result:
o The DR3900 will display the High-Level Iron (HLI) concentration in ppm
(mg/L).

🔹 Low-Level Iron (LLI) Test Procedure

APPARATUS:

• 10 mL Square cell
• 50 mL Beakers (x2)
• Laboratory Oven (Capable of 90°C)

REAGENTS:

• Iron Indicator (Fe-4 Reagent)

PROCEDURE:

1. Prepare the Samples:

o Pour 50 mL of the water sample into one beaker as shown in figure.
o Pour 50 mL of distilled water into a second beaker (for blank) as shown in
figure.
2. Add Reagent:
o Add 2 mL of Iron Indicator (Fe-4 Reagent) into each beaker as shown in
figure.
o Stir gently to mix
3. Heat the Solutions:
o Place both beakers in a laboratory oven.
o Heat at 90°C for 40 minutes.
4. Cool and Transfer:
o After heating, allow both beakers to cool slightly.
o Take 10 mL from each beaker and pour into separate clean 10 mL cell.
 5. Zero the Instrument:
o Insert the cell containing distilled water + reagent into the spectrophotometer.
o Press “Zero” to calibrate.
6. Measure the Sample:
o Insert the cell containing the sample + reagent.
o Press “Read” to take the measurement.
7. Record the Result:
o The DR3900 will display the Low-Level Iron (LLI) concentration in ppm
(mg/L).

Notes:
• Ensure beakers are heat-resistant and cleaned before use.
• Do not skip the blank (distilled water) step — it's essential for accurate results.
• Always use gloves and safety gear when handling hot samples and reagents.
• Let cell cool to room temperature before reading if they're too warm.

🔹 Nitrate Test (Titration Method)

APPARATUS:

• 50 mL Beaker
• Dropper (for reagents)

REAGENTS:

• R34 – Ferroin Indicator

• R35 – Marine Indicator (Titrant)

PROCEDURE:

1. Prepare the Sample:

o Take 2 mL of the water sample in a clean 50 mL beaker.
2. Add Indicator:
o Add 4 drops of R34 (Ferroin Indicator) to the sample.
o A red color will appear as shown in figure 1, indicating the starting point of
the reaction.
3. Titrate the Sample:
Figure 1
 o Slowly add R35 (Marine Indicator) drop by drop.
o Stir gently after each drop.
o Continue adding R35 until the solution changes from red to parrot green as
shown in figure 2.
4. Calculate the Nitrate Concentration:
o Count the number of drops of R35 used.
o Multiply the number of drops by 50 to get the Nitrate concentration in ppm.
Figure 2
Formula:
Nitrate (ppm) = No. of R35 drops × 50

Notes:
• Add R35 slowly and stir between drops to avoid overshooting the color change.
• Perform the test under good lighting to observe the color shift accurately.
• Use clean droppers for each reagent to avoid cross-contamination.
• This is a manual test — no spectrophotometer is required.

Muhammad Ahsan

Cell # : 03455102241

#OPEN TO WORK