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Saturday, October 18, 2008

CALIBRATION OF UV-VISIBLE SPECTROPHOTOMETER

1.0 CONTROL OF ABSORBANCES
PROCEDURE: REPARATION OF POTASSIUM DICROMATE SOLUTION: Dry K2Cr2O7 at 130oC for 2 hours. Weigh accurately about the quantity of sample equal to 60mg into 1000ml volumetric flask. Add 0.005m Sulphuric acid to dissolve and dilute with the same solvent.
Find out the absorbances of (A 1% 1cm) above said solution at wavelengths of a 235nm, 257nm, 313nm and 350nm. Calculate the (1% 1cm) value using the following expression.
Absorbance X 1 X 1000 / Weight of K2Cr2O7 in g X 100
Check the value for each wavelength with the tolerances given four wavelengths.


PREPARATION OF REAGENT:
(i) Dilute 3ml of concentrated sulphuric acid and make up 100ml with distilled water.
(ii) Dilute 20ml to 2000ml with distilled water.
2. LIMIT OF STRAYLIGHT:
The absorbance of a 1.2-% w/v solution of potassium chloride at a path-length of 1cm should be greater than 2.0 at about 200nm when compared with water as reference liquid.
1.2% POTASSIUM CHLORIDE SOLUTION:
Take 1.2 g of Potassium chloride previously dried at 105oC for two hours into the 100ml volumetric flask. Add water to dissolve and make up to volume with water.
3. RESOLUTION POWER:
Record the spectrum of a 0.02% v/v solution of Toluene in Hexane UV. The ratio of the absorbance at the maximum at about 269nm to that at the minimum at about 266nm is not less than 1.5 unless other wise specified in the monograph.
0.02% v/v TOLUENE:
Take 0.02ml of Toluene into the 100ml volumetric flask. Add Hexane to volume.

26 comments:

eswar said...

Dear sir/Madam
Your Artical for calibration procedure for UV/VISIBLE SPECTROPHOTOMETER is very excelent and very usuful forme.
and another request please send calibration procedure for HPLC and GC.

Regards,
eswar.

peri said...

Dear Sir/madam
Why we have to use Potassium Dichromate and Sulphuric acid for uv calibration?
Is there any reason to heat the Potassium Dichromate at 130C?

hari said...

Dear sir/Madam
there is any importance usin Sulphuric acid(0.005M) as a medium for Dichormate in UV Claibration,
This UV Calibration is very useful to me plz provide GC calibration for me
Thanking you

Regards,
SANKAR,

harisankar said...

Dear sir/Madam
Please Provide Gc calibration for me this UV calibration is very useful for me
there is any importace Using Sulphuric acid(0.005M) as a Diluent for Dichormate
Thanking you

jengiti said...

dear sir/madam,

Your article is very use ful and provide, why k2Cr2O7 is only used for calibration of UV in limit of stray light
thanking you,

chintu said...

Dear sir/madam
Why we have to use particula chemicals(eg-Pot.dichromate,pot chloride,0.005M sulphuric acid etc)in calibration of UV spectrophotometer,pls send me mail

regards,
Nagaraj.P.E

kndprasad said...

Dear Sir/madam
Why we have to use Potassium Dichromate and Sulphuric acid for uv calibration?
Is there any reason to heat the Potassium Dichromate at 130C?

kndprasad said...

Dear sir/madam
Why we have to use particula chemicals(eg-Pot.dichromate,pot chloride,0.005M sulphuric acid etc)in calibration of UV spectrophotometer,pls send me mail

kndprasad said...

Dear sir/Madam
Your Artical for calibration procedure for UV/VISIBLE SPECTROPHOTOMETER is very excelent and very usuful forme.
and another request why we use deuterium in uv lamp

PRADEEP said...

Dear sir/madam
Why we have to use particular chemicals(eg-Pot.dichromate,potassium chloride,0.005M sulphuric acid etc)in calibration of UV spectrophotometer,pls send me mail

regards,
PRADEEP (UP)

latha said...

DEAR SIR/MADAM
Your Artical for calibration procedure for UV/VISIBLE SPECTROPHOTOMETER is very excelent and very usuful forme.
and another request please send Validation of uv/visible spectrophotometer
Thanking you

Regards,
latha,

pankaj singh said...

Dear Sir/madam
Why we have to use Potassium Dichromate and Sulphuric acid for uv calibration?
please send me s.o.p for shimadzu u.v.spectrophotometer.
thanking you
pankaj singh patel
e.mail-pankajsinghpatel@gmail.com

jyoti said...

Dear sir/madam,

this article is very use full for me please provide me why only this chemicals like K2Cr2O7,hexane and toluene used in calibration, why not other?
thanking you,

neelam said...

dear sir/madam
why we use 4 % holmium oxide in 1.4 M perchloric acid while mentioned in IP is 5 %.and it is not giving correct result, please help me to prepare this

siti fatimah said...

How do you determine the limit for the resolution (minimum ratio of the absorbance at the 269nm to that at the minimum at 266nm of a 0.02 percent V/V solution of toluene R in hexane R) as not less than 1.5.Is it based on trending?

requistation said...

Dear sir/Madam
Your Artical for calibration procedure for UV/VISIBLE SPECTROPHOTOMETER is very excelent and very usuful for me.
and another request please send calibration procedure for HPLC and GC.

Regards,
viswa
(chennai)

requistation said...

Dear sir/madam
Why we have to use particular chemicals(eg-Pot.dichromate,potassium chloride,0.005M sulphuric acid etc)in calibration of UV spectrophotometer,pls send me mail

rgards
viswa

requistation said...

Dear sir/madam
Why we have to use particular chemicals(eg-Pot.dichromate,potassium chloride,0.005M sulphuric acid etc)in calibration of UV spectrophotometer,pls send me mail

requistation said...

Dear sir/madam
Why we have to use particular chemicals(eg-Pot.dichromate,potassium chloride,0.005M sulphuric acid etc)in calibration of UV spectrophotometer,pls send me mail


regards
viswa

amit kumar said...

Dear Sir,
please tell me the procedure of "control of wavelength" in UV-spectrophotometer calibration.


Regards,
Amit Kumart

Bipin said...

Respected seniors;
Why we used 0.02% toluene in hexane.

hari priya said...

Hari Priya said: Dear sir/Madam What is stray light? Why we have to use Potassium Dichromate and Sulphuric acid for uv calibration? Regards,
Priya.

Nilesh Disale said...

Dear sir/Madam
Please Provide Gc calibration for me this UV calibration is very useful for me
there is any importace Using Sulphuric acid(0.005M) as a Diluent for Dichormate
Thanking you

Nilesh Disale said...

thank u...............

Anonymous said...

why kcl,k2cr207 shall be dried at 105C For one hour before calibration IN UV

chethan said...

Y 60 mg K2Cr2O7 used