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Calibration of Uv Visible Spectrophotometer

Calibration involves ensuring that a measurement method or instrument will produce accurate results using a primary reference standard. It should be performed when a specific time period or operating hours has elapsed, with a new instrument, or when observations appear questionable. Calibration involves controlling the wavelength and absorbance, maintaining the instrument, taking precautions, and understanding abbreviations. When the instrument is not producing calibrated results, it should be labeled as faulty and repaired by a service engineer. The instrument should be kept free from dust, magnetic/electric fields, vibrations, and direct sunlight to extend its life.

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1K views

Calibration of Uv Visible Spectrophotometer

Calibration involves ensuring that a measurement method or instrument will produce accurate results using a primary reference standard. It should be performed when a specific time period or operating hours has elapsed, with a new instrument, or when observations appear questionable. Calibration involves controlling the wavelength and absorbance, maintaining the instrument, taking precautions, and understanding abbreviations. When the instrument is not producing calibrated results, it should be labeled as faulty and repaired by a service engineer. The instrument should be kept free from dust, magnetic/electric fields, vibrations, and direct sunlight to extend its life.

Uploaded by

ujjwal
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
Available Formats
Download as PPTX, PDF, TXT or read online on Scribd
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Unit 5

What is calibration:
• Calibration is the act of ensuring that a method or
instrument used in measurement will produce
accurate results.
• Calibration is performed using primary reference
standard
 When a specific time period is elapsed.
 When a specific operating hours has elapsed.

 With a new instrument.

 When observations appear questionable.


1. Control of wavelength
2. Control of absorbance
For ultraviolet region
For visible region
Limit of stray light
Resolution power
3. Maintenance
4. Precautions
5. Abbrevations
STEP 1: Dissolve 1
STEP 2: Record the
gram of holmium
CHEMICALS: 1.4M spectrum
oxide in 1.4M
perchloric acid, holmium
perchloric acid
perchlorate
holmium with the aid of
solution from
perchlorate heating on water
200nm to 600nm
solution bath, cool and
using 1.4M
dilute to 25ml with
perchloric acid as
same solvent.
reference solution
 Note down the maxima
observed at wavelength
against the acceptance criteria
given below:
 Deviation of wavelength reading at an
absorbance band
STEP 1:Weigh 57 to 63mg of potassium
dichromate primary standard and Step 2: Measure the absorbance at 235
transfer to 1000ml volumetric flask. nm, 257nm, 313nm and 350nm using
Dissolve in 0.005m sulphuric acid and 0.005M sulphuric acid as reference
make up to the mark with the same
acid.
 FOR VISIBLE REGION: Whole procedure is
same as UV region but at the end measure the
absorbance at 430nm.
 CALCULATION: Value of A(1%,1cm)

A(1%,1cm)= absorbance x 100


weight in gm x 100
STEP 1: Prepare the solution of
1.2%v/v potassium chloride and
dissolve with 50ml distilled water.

STEP 2:Determine the absorbance


at 200nm against purified water as
blank
STEP 1: Prepare a solution 0.02%v/v toluene in hexane

STEP 2: Record the spectrum of 0.02%v/v toluene in hexane from


250nm – 300nm using hexane as reference

STEP 3: record the absorbance at 269nm (max) and 266nm (min)n

STEP 4: Calculate the ratio of absorbance by dividing the absorbance


at maxima and minima
MAINTAINENCE
When the instrument not produce calibrated

results it should be labeled “FAUL TY” and should

be repaired by service engineer of instrument.


 The instrument should be always free from dust

 Place the instrument as far as possible from any strong magnetic or


electric field or any electric apparatus generating a high frequency.

 Avoid places where vibrations exist or direct sunlight.

 To extent the life of source lamp have them powered only when
required.

 The temp. Of all solution used in the test should not differ by more then
0.5ºC.

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