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TP and Calibration

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TP and Calibration

Uploaded by

Khairi Ali Omar
Copyright
© © All Rights Reserved
Available Formats
Download as PDF, TXT or read online on Scribd
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A P P L I C AT I O N N O T E

UV/Visible Spectroscopy

Water Analysis using


LAMBDA: Total Phosphorus Introduction

(T-P), Ascorbic Acid Method In this application


note, the quantitative
analysis of total
phosphorus (T-P)
was performed by as-corbic acid method. Data are rapidly acquired
using LAMBDA™ 465 UV-Vis Spectrophotometer and processed using
UV Lab™ Software.

Principle
Total phosphoric compounds in water sample is changed to
phosphate (PO4-) form by oxidation. After treatment with ammonium
molybdate • ascorbic acid solution, blue color is created. This color
is measured at 880 nm.
Reagents and Apparatus
1. Phosphate ion standard solution (0.005 mg P/mL)
2. Unknown sample
3. Ammonium molybdate solution
-D
 issolve 6 g ammonium molybdate tetrahydrate
((NH4)6Mo7O24·4H2O) and 0.24 g antimonyl potassium
tartrate (C4H4O6K·SbO·1/2H2O) in 300 mL D.I water, add
120 mL sulfuric acid (2 + 1) and 5 g ammonium sulfamate
(NH4OSO2 NH2), dilute to be 500 mL with D.I water.
4. 7.2% L-Ascorbic acid solution
- Dissolve 7.2 g L-ascorbic acid (C6H8O6) in 100 mL D.I water.
5. Ammonium molybdate • Ascorbic acid solution
- 500 mL ammonium molybdate solution mix with 100 mL
7.2% L-ascorbic acid solution.
6. LAMBDA 465 (PDA UV/Vis Spectrophotometer)
7. UV Lab Software
8. Cuvettes (10 mm pathlength) Figure 1. Experimental setup for T-P analysis.

Procedure Result
1. Prepare serial volume (1~20 mL) of phosphate ion standard 1. Calibration curve
solution (0.005 mg P/mL) in 100 mL volumetric flasks for
standards, dilute to volume with D.I water. Fill 25 mL nessler Figure 2 shows spectra of T-P standards. Table 1 and Figure 3
tubes with them each, then perform the experiment as show data and calibration curve of six standards. Correlation
following procedure. coefficient R2 is 0.9962.

2. Fill a 25 mL nessler tube with unknown sample. Table 1. Calibration data of T-P standards.

3. Add 2 mL ammonium molybdate • ascorbic acid solution. Concentration AU


No. Name
(ppm) (880.00 nm)
4. Mix and leave for 15 min at 20~40 °C.
1 Standard 1 0.00 0.0005
5. In Quantification Standard mode, measure the absorbance 2 Standard 2 0.10 0.0671
of standards with reference to standard 1 (0 ppm) at 880 nm.
3 Standard 3 0.20 0.1232
6. In Quantification Sample mode, measure the absorbance of 4 Standard 4 0.50 0.3102
unknown sample and calculate its concentration.
5 Standard 5 1.00 0.6068
Instrument Parameters 6 Standard 6 2.00 01.0829

The instrument parameters of the LAMBDA 465 are as follows: R2 = 0.99620


Figure 1 shows experimental setup. Function : Y = 0.5432X + 0.0211

Experiment Setup 2. Unknown sample


Data type: Absorbance
Concentration of unknown sample was acquired using
Sampling: Single cell
calibration curve of Figure 3. The concentration of unknown
Mode: (Spectra no.: 1/Scan no.: 30/
sample is 0.53 ppm (see Table 2).
Integration no.: 1/Gain no.: 1)
Experiment Method Table 2. Concentration of unknown sample.
Use wavelength: 880 nm Concentration Dilution AU
Name
Curve dimension: 1 (ppm) Factor (880.00 nm)
Sample 1 0.53 1.0 0.3085

2
Conclusion
Using LAMBDA 465 and UV Lab Software, quantitative analysis
of total phosphorus (T-P) in water was performed. Rapid
acquirement of spectra and good sensitivity were obtained with
LAMBDA 465. Good calibration curve of which R2 is 0.9962 was
acquired. UV Lab Software was used effectively for quantitative
analysis and to process the data efficiently.

Figure 2. The spectra of T-P standards by ascorbic acid method.

Figure 3. The calibration curve of T-P standards.

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