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Assay of Sulphanilamide

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Sulphonamide/Sulfonamide

Amide Sulphone Sulphonamide

A functional group for most sulphur/sulpha drugs

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Some Sulphonamide Drugs

Hydrochlorothiazide Furosemide Sulfamethoxazole

(Diuretic) (Diuretic) (Antibiotic)

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Sulfa/Sulpha drugs

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Sulfonamides
• The term has become more associated with
antibiotics which are structurally similar with
p-aminobenzoic acid (PABA).
• Largely bacteriostatic against gram +ve and
gram –ve bacteria.
• Eg. Sulfadiazine, sulphamethoxazole,
cotrimoxazole, trimethoprim.

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Sulphanilamide – A Sulphur drug

• 1932 – Gerhard Domagk observe the effect of

Prontosil on Streptococcus in mice.
• A sulphonamide (para-aminobenzenesulfonamide)
is the API.
• Magic bullet of the WWII; standard in 1st aid kits.
• Used to be very useful but currently only indicated
for vaginal yeast infections.

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Sulphanilamide
• Antibiotic – antibacterial agents (kills bacteria).
• General mechanism: interference with some
normal process in the organism.

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Assay of Sulphanilamide
Assay of Sulphanilamide with NaNO2 (1 : 1)
Step 1
Standardization of 0.1 M NaNO2 solution with 0.02 M
KMnO4 solution

Step 2
Reaction of sulphanilamide with 00.01 M NaNO2
solution.
**KMnO4 is not a primary standard!

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Standardization of 0.1 M NaNO2(aq)

solution with 0.02 M KMnO4(aq) solution
Overall Reaction 1
2MnO4- (aq) + 5NO2- (aq) + 6H+ (aq)  2Mn2+ (aq) + 5NO3- (aq) + 3H2O (l)

Generation of NO2- (aq) in acidic environment

2NaNO2 (aq) + H2SO4 (aq)  Na2SO4 (aq) + 2HNO2 (aq)

3HNO2 (aq)  HNO3 (aq) +H2O (l) + 2NO (g) ↑

Breakdown of Nitrous acid

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Standardization of 0.1 M NaNO2(aq)

solution with 0.02 M KMnO4(aq) solution
Into a conical flask pipette 0.02 M
KMnO4 solution (25 mL) and add
water (about 30 mL). Add conc. H2SO4
0.1 M NaNO2
(4 - 5 mL) rotating the flask as the
solution
addition is made. Titrate with the
NaNO2(aq) solution in a continuous
stream of drops, shaking and rotating
the flask vigorously and continuously.
1 mL 0.1 M NaNO2  2 mL 0.02 M (0.1 N)
0.02 M KMnO4
KMnO4
Solution
acidified with Expected endpoint = 12.25 mL
H2SO4(aq) f(NaNO2)= Actual/ Nominal

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Reaction of sulphanilamide with

0.1 M NaNO2 solution.
• Primary amine reacts with NaNO2 solution to yield
diazonium salt.

0.1 M NaNO2
solution
Add
1. 2.5000 g of C6H8N2SO2.
2. 50 mL conc. HCl.
3. 5 g KBr dissolved in H2O.
4. Make up to volume.

25.00 mL aliquot
250 mL Flask
0.017221 g NH2C6H4NH2  1 mL 0.1 M NaNO2
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Mechanism of End Point

N02- (aq) + I- (aq) + H+ (aq) → H2O (l) + NO (g) + 1/2 I2(aq)

Iodine reacts with starch to yield a dark blue/purple

colour

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Determination of %W/V
• How much sulphanilamide is in 25 mL and hence
100 mL (mass in 100 mL of solution)

If 0.1 M NaNO2 that reacted is x mL

0.017221 g NH2C6H4NH2  1 mL 0.1 M NaNO2

In 25 mL contains (0.0177221 g × X × f )
%w/v = (0.0177221 g × X × f ) *100/25

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Determination of % content
• Assuming the sample was obtained from tablet
• An equivalent weight would have been taken.
• Estimate equivalent weight as y g.

API in 25 mL, m1 = (0.0177221 g × x )

Mass of y in 25 mL, m2 = y × 25 mL/250 mL

%w/w = m1/m2 × 100%

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Percentage Purity
• Percentage purity determines how pure an API is.
• This is also referred to as Assay.

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Considerations for Titrations

• Three burette readings required and must not be
more than 0.05 mL apart from each other for both
standardization and assay of sulphanilamide.

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Precautions
• Appropriate burette reading.
• Good pipetting and solution transfer skills.
• Avoid spilling of reagents.
• Ensure proper mixing of all solutions.

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Think through
• You have an analyte A that you want to assay.
• You A reacts with B in a 1 :3 ratio
• B can be standardize with C and their
stoichiometry is 4:5
• What concentrations of B and C will u prepare and
why?

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