USP 2023: SULFAMETHOXAZOLE & TRIMETHOPRIM (COTRIMOXAZOLE)

TABLETS MONOGRAPH
A. DEFINITION
Sulfamethoxazole and Trimethoprim Tablets contain not less than (NLT) 93.0% and not more
than (NMT) 107.0% of the labeled amounts of Sulfamethoxazole & Trimethoprim.
Dissolution specification: NLT 75% of the labeled amount of Sulfamethoxazole and
Trimethoprim is dissolved after 60 minutes.
B. IDENTIFICATION
The retention time of the peak of Sulfamethoxazole and Trimethoprim of the Standard solution
correspond to that of the sample solution as obtained in the Assay.
C. ASSAY
Procedure
Mobile phase: Mix 1400 mL of water, 400 mL of acetonitrile and 2.0 mL of trimethylamine in a
2000 mL volumetric flask. Allow to equilibrate to room temperature, and adjust pH to 5.9 ± 0.1
with 0.2N sodium hydroxide or dilute glacial acetic acid. Then dilute with water to volume, pass
through a filter of 0.45 μm.
Standard solution: Prepare Trimethoprim reference/working standard to obtain a concentration
of 0.032 mg/mL and 0.032j mg/mL of Sulfamethoxazole in small amount of methanol and then
top up with the mobile phase. j is the ratio of the labeled amount in mg of Sulfamethoxazole to
the labeled amount in mg of Trimethoprim.
Sample solution: Transfer from finely powdered Tablets (NLT 20), an equivalent to 160 mg of
Trimethoprim, to a 100 mL volumetric flask. Add 50 mL of methanol and sonicate, with
intermittent shaking, for 5 minutes. Allow to equilibrate to room temperature and dilute with
methanol to volume, then filter. Dilute further using mobile phase so as to obtain a concentration
of 0.032 mg/mL of the Trimethoprim.
Chromatographic conditions
a. Mode: Liquid chromatography (LC)
b. Detector: UV at 254 nm.
c. Column: 25cm × 4.6 mm, 5 μm
d. Flow rate: 1-2 mL/min.
e. Injection volume: 20 μL

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D. DISSOLUTION
Medium: 0.1N HCl, 900 mL
Apparatus: 2, 75 rpm
Time: 60 minutes
For the chromatographic conditions and the standard concentration proceed as in Assay.
A Pharmacist was provided with six batches of Cotrimoxazole 960 mg tablets (each tablet
contains Sulfamethoxazole BP 800 mg and Trimethoprim BP 160 mg) for analysis in the Quality
Control Laboratory. An analyst carried out the assay and dissolution according to the USP
monograph. The analysis of one of the batches generated the following data.
Tablet number Weight (mg)
1 1118.28
2 1091.09
3 1093.13
4 1077.25
5 1092.82
6 1090.77
7 1079.71
8 1101.27
9 1081.20
10 1057.88
11 1083.46
12 1067.36
13 1073.38
14 1097.86
15 1088.89
16 1090.49
17 1076.98
18 1080.11
19 1055.65
20 1072.05

a. Calculate the average weight of Cotrimoxazole Tablets.

b. Calculate the percentage deviation from the mean of each tablet

c. Calculate the upper and lower acceptable deviation from the mean weight.

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 TRIMETHOPRIM
Standard A Standard B Sample A Sample B Sample C Dissolution
Peak 3025304 2804627 2696894 2754737 2280735 2613240
Area/Response
3035530 2792114 2600561 2751099 2501385 2609802
3039400 2790470 2703913 2747391 2704110 2608085

Weights (mg) 21.27 19.96 1080.07 1091.12 1090.68 2608022

Desired 2604525
Concentration
0.032 2601981
(mg/mL)
Dilution factor 625 5000 4500
(ml)
Standard Assay on drier basis 99.3%
Loss on drying (LOD) 0.3% 99.3% - 0.3% = 99.0%

SULFAMETHOXAZOLE
Standard A Standard B Sample A Sample B Sample C Dissolution
Response 43249806 46798853 37565106 38354680 31719926 36675227
43438191 46644891 37752864 38261955 34804823 36647860
43453342 46681899 37648341 38246300 37621600 36643188

Weights (mg) 17.68 18.86 1080.07 1091.12 1090.68 36633957

Desired 36589759
concentration
0.16 36600410
(mg/mL)
Dilution factor 100 5000 4500
(mL)
Standard Assay on dried basis 99.28%
Loss on drying 0.26% 99.28% - 0.26% = 99.02%

d. Calculate desired standard weight.

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e. Calculate the actual standard concentration.
f. Calculate the standard normalized response.

g. Calculate the percentage amount of Trimethoprim (and Sulfamethoxazole) in the sample.

ii. Determine the amount of Trimethoprim (and Sulfamethoxazole) in the sample powder taken.

iii. Determine the amount of Trimethoprim (and Sulfamethoxazole) in one tablet.

iv. Determine the percentage content/label claim and hence the average percentage for each
molecule.
v. What conclusion do you make from the sample of Cotrimoxazole Tablets provided for the
assay?

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