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Titration Method

The document outlines a titration method for estimating amino acids, specifically glycine, using formaldehyde and sodium hydroxide. It details the preparation of reagents, the procedure for titration, and the calculations required to determine the concentration of unknown glycine solutions. The method relies on the reaction of amino acids with formaldehyde and the subsequent titration with sodium hydroxide to achieve accurate results.

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19 views

Titration Method

The document outlines a titration method for estimating amino acids, specifically glycine, using formaldehyde and sodium hydroxide. It details the preparation of reagents, the procedure for titration, and the calculations required to determine the concentration of unknown glycine solutions. The method relies on the reaction of amino acids with formaldehyde and the subsequent titration with sodium hydroxide to achieve accurate results.

Uploaded by

surbhisaini0904
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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Experiment-6: Titration Methods for Estimation of Amino

acids
Aim: Estimation of Amino acids by Titration methods

Principle:

Amino acids react with formalin to from methylene amino acids. But it was found that
the reaction was more complex and the main product is monomethylol and di
methylol amino acids. Amino acids exist in zwitter ionic form and cannot be titrated
directly with alkali. Thus amino groups of amino acids are blocked by reaction with
formaldehyde. But formaldehyde does not react with the charged amino groups
(-NH3+), thus first the amino acid reacts with sodium hydroxide solution to give
glycine which condenses with formaldehyde to give a stable anion.
Further formaldehyde solution contains formic acid and also amino acids are not
exactly neutral species, thus it is necessary that both the formaldehyde and amino acid
solution should have the same pH before mixing and for this purpose each solution is
first made just alkaline to phenolphthalein by means of dilute sodium hydroxide
solution.

Reagents:
a. Neutral formalin solution
It is prepared by neutralization of formalin solution with alkali to
phenolphthalein end point.
b. Standard Glycine solution
An accurately weighed 0.8 grams of Glycine is dissolved in distilled water in a
measuring flask 100 mL.
c. Unknown Glycine solution
d. Sodium hydroxide solution 0.1 N
e. Phenolphthalein indicator

Procedure:
1. A standard glycine solution (20 mL) is pipetted out into a conical flask and
phenolphthalein (2-3 drops) added.
2. It is titrated with sodium hydroxide solution until a faint pink color appears.
3. Then the neutralized formaldehyde solution (10 mL) is added.
4. The pink color of the solution in the titration flask disappears immediately and
the solution becomes acidic.
5. Then the contents of the flasks are titrated with sodium hydroxide solution till
the pink color is obtained.
6. Same experiment repeated with unknown glycine solution.

Observations:
Weight of Glycine dissolved in 100 mL = w g
Volume of sodium hydroxide solution used with standard glycine solution=v1 mL
Volume of sodium hydroxide solution used with unknown glycine solution=v2 mL
Volume of glycine solution taken= 20 mL

1
Calculations

V1 mL of NaOH in 20mL standard glycine solution= (w/5) g of glycine


V2mL of Na OH in 20mL of unknown glycine solution= (w/5) x (v2/v1) g of glycine
Thus strength= (w/5) (v2/v1) (1000/20) g/L
= (wxv2x10)/v1 g/L

Result:

2
Reference:

1) Sadasivam S and Balasubramanian T (1985). Practical Manual


(Undergraduate), Tamil Nadu Agriculture University, Coimbatore, p.2.

2) Sadasivam S, Manickam A (2008). Biochemical Methods, New Age


International Publishers, 3rd Edition, ISBN: 978-81-224-2140-8.

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