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Swami Vivekanand College of Pharmacy, Indore

This document provides instructions for performing the assay of Dextrose Injection I.P. using colorimetry. It describes the required glassware, chemicals including the DNSA solution, and the colorimetric principle where dextrose produces an intense red color with DNSA in alkaline medium. The procedure involves preparing standard glucose solutions, adding DNSA and heating to develop color, then measuring absorbance to construct a standard curve and determine the unknown dextrose concentration.

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Shubham wani
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3K views

Swami Vivekanand College of Pharmacy, Indore

This document provides instructions for performing the assay of Dextrose Injection I.P. using colorimetry. It describes the required glassware, chemicals including the DNSA solution, and the colorimetric principle where dextrose produces an intense red color with DNSA in alkaline medium. The procedure involves preparing standard glucose solutions, adding DNSA and heating to develop color, then measuring absorbance to construct a standard curve and determine the unknown dextrose concentration.

Uploaded by

Shubham wani
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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Download as PDF, TXT or read online on Scribd
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SWAMI VIVEKANAND COLLEGE OF PHARMACY,INDORE

Experiment no. 3
Object- To perform the assay of Dextrose Injection I.P. by Colorimetry
Requirements-
Glassware required- Pipettes(1ml,5ml,10ml), Beakers(250ml), vol.
flask(100ml) Test tubes, testtube stand.
Chemical required-Std. glucose solution-300mg, 3N NaOH, Rochelle
salt -150gm, DNSA solution
DNSA solution-Take 5gm of DNSA→In 100ml of 2 N
NaOH→Dissolve and add 250 ml of distilled water and mix→Add 150
gm of Rochelle salt(sod. Pot. tartrate)→dissolve and make up the vol. to
500ml with distilled water→ filter if necessory→ stored in a amber
coloured glass bottle.
Principle-
Dextrose gives an intense red color with di nitro salicylic acid (DNSA)
in an alkaline medium . The reduced DNSA formed, has maximum
absorbance at 600nm. Estimation , hence involves color development
followed by reading the A using a suitable colorimeter against a reagent
black. A std plot with known concentration of glucose is first made from
which known conc. Can be easily found out.
Operation-
(1) Turn on the colorimeter warm up for 15 min.
(2) Set zero.
(3) Set wavelength.
(4) Insert blank
(5) Set full scale.
(6) Insert unknown.
SWAMI VIVEKANAND COLLEGE OF PHARMACY,INDORE

(7) Read % T or A.

Procedure-
(1) Prepare glucose solution (stock) 300mg%.
(2) Pipette 0,0.5,1.0,2.0 and 2.5 ml of above solution in to
clean corning test tubes of about 20ml of capacity.
(3) Add 3 ml of 3N NaOH to all the test tubes.
(4) Add 3ml of DNSA solution solution to all the test tubes.
(5) Add enough distilled water to make 10ml in each
case.(4ml,3.5ml.3ml,2.5ml,2.0ml,1.5ml)
(6) Shake the test tubes to mix contents.
(7) Keep the test tubes in boiling water for 15 min. cool the
test tubes.
(8) Read A or %T at 600nm.
(9) Dextrose inj. I.P.(25%w/v). Dilute 1 ml to 100ml. Take 1
ml and 2ml of diluted sample. Proceed from step (3) of
procedure.
Observations-
Conc. of standard stock Glucose solution -300mg%
Sr. no. Stock Conc. A %T
sample µg/ml
1. 0.0 ml -
2. 0.5 ml 150
3. 1.0 ml 300
4. 1.5 ml 450
5. 2.0 ml 600
SWAMI VIVEKANAND COLLEGE OF PHARMACY,INDORE

6. 2.5 ml 750
7. Unknown

Calculation-
Result-
Given unknown solution contain------------- mg/ml of Glucose.

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