Perindopril Erbumine
C23H43N3O5 Mol. Wt. 441.6
Perindopril Erbumine is 2-methylpropan-2-amine(2S,3aS,7aS)-1-[(2S)-2-[[(1S)-1(ethoxycarbonyl)butyl]amino]pro panoyl]octahydro-1H-indole-2-carboxylate.
Perindopril Erbumine contains not less than 99.0 per cent and not more than 101.0 per cent of C23H43N3O5, calculated on the anhydrous basis.
Category. Antihypertensive.
Dose. Initial dose: 4 mg daily for a weak, then increase to maintenance dose as tolerated.
Maintenance dose: 8 mg once a day.
Description. A white or almost white, slightly hygroscopic, crystalline powder.
Identification
- Determine by infrared absorption spectrophotometry (2.4.6). Compare the spectrum with that obtained withPerindoprilerbumine RS or with the reference spectrum of perindopril erbumine.
If the spectra obtained show differences, dissolve the substance under examination and the reference substance separately in methylene chloride, evaporate to dryness and record new spectra using the residues.
- In the perindopril impurity A, the principal spot in the chromatogram obtained with the test solution corresponding to the spot in the chromatogram obtained with reference solution (c).
Tests
Specific Optical rotation (2.4.22). -69° to -66° (anhydrous basic), determined on 1.0 per cent w/v solution in ethanol (95 per cent).
Impurity A. Determine by thin-layer chromatography (2.4.17), coating the plate with silica gel.
Mobile phase. A mixture of 1 volume of glacial acetic acid, 40 volumes of toluene and 60 volumes of methanol.
Test solution. Dissolve 200 mg of the substance under examination in methanol and dilute to 10.0 ml with the same solvent.
Reference solution (a). Dissolve 5 mg of perindopril impurity A RS (2S,3aS,7aS)-octahydro-1H-indole-2-carboxylic acid) in methanol and dilute to 25.0 ml with the same solvent.
Reference solution (b). Dilute 5.0 ml of reference solution (a) to 20.0 ml with methanol.
Reference solution (c). To 5.0 ml of reference solution (a), add 5 ml of a 2 per cent w/v solution of 1,1-dimethylethylamine in methanol.
Apply to the plate 10 µl of the test solution, reference solution (b) and reference solution (c). Dry the plate in a current of warm air and expose to iodine vapour for at least 20 hours. The chromatogram obtained with reference solution (c) shows two clearly separated spots. Any spot due to perindopril impurity A is not more intense than the spot in the chromatogram obtained with reference solution (b) (0.25 per cent).
Stereochemical purity. Determine by liquid chromatography (2.4.14).
Test solution. Dissolve 20 mg of the substance under examination in ethanol (95 per cent) and dilute to 10.0 ml with the same solvent.
Reference solution (a). Dilute 1.0 ml of the test solution to 100.0 ml with ethanol (95 per cent) . Dilute 1.0 ml of this solution to 10.0 ml with the same solvent.
Reference solution (b). Dissolve 10 mg of perindopril for stereochemical purity RS (containing perindopril impurity I (2RS,3aRS,7aRS)-1-[(2RS)-2-[[(1SR)-1-(ethoxycarbonyl)butyl]amino]propanoyl]octahydro-1H-indole-2-carboxylic acid ((±)-1′-epi-perindopril)) in ethanol (95 per cent) R and dilute to 5.0 ml with the same solvent.
Chromatographic system
– a stainless steel column 25 cm x 4.6 mm, packed with spherical octadecylsilane bonded to porous silica
(5 µm),
– column temperature: 50°, for the column and the tubing preceding the column (the method has been developed with a temperature of 50° for at least 30 cm of the tubing preceding the column).
– mobile phase: a mixture of 21.7 volumes of acetonitrile, 0.3 volumes of pentanol and 78 volumes of a 0.15 per cent w/v solution of sodium heptanesulphonate previously adjusted to pH 2.0 with a mixture of equal volumes of perchloric acid and water,
– flow rate: 0.8 ml per minute,
– spectrophotometer set at 215 nm,
– equilibration time 240 minutes,
– injection volume: 10 µl.
Run the chromatogram 1.5 times the retention time of principal peak.
The relative retention time with reference to perindopril (retention time = about 100 min): perindopril impurity I = about 0.9.
Inject reference solution (b). The chromatogram obtained with reference solution (b) is similar to the chromatogram supplied with perindopril for stereochemical purity RS.
Inject reference solution (a). The signal to noise ratio is not less than 3 for the principal peak.
Inject reference solution (b) the peak to valley ratio is not less than 3, where Hp = height above the baseline of the peak due to perindopril impurity I and Hv = height above the baseline of the lowest point of the curve separating this peak from the peak due to perindopril.
Inject reference solution (a) and the test solution. In the chromatogram obtained with the test solution, the area of perindopril impurity I is not more than the principal peak in the chromatogram obtained with reference solution (a) (0.1 per cent). The area of any secondary peak for each impurity is not more than the area of the principal peak obtained with reference solution (a) (0.1 per cent). Ignore any peak with a relative retention with reference to the principal peak of less than 0.6 or more than 1.4.
Related substances. Determine by liquid chromatography (2.4.14).
Note: Prepare the solutions immediately before use or maintain at a temperature below 10 °.
Test solution: Dissolve 60 mg of the substance under examination in mobile phase A and dilute to 20.0 ml with mobile phase A.
Reference solution (a): Dissolve 3 mg of perindopril for peak identification RS (containing impurities B, E, F, H and K) in 1 ml of mobile phase A.
Reference solution (b): Dilute 1.0 ml of the test solution to 200.0 ml with mobile phase A.
Reference solution (c): Dilute 1.0 ml of reference solution (b) to 10.0 ml with mobile phase A.
Chromatographic system
– a stainless steel column 15 cm x 4.0 mm, spherical end-capped octylsilane bonded to porous silica (5 µm),
– column temperature: 60°, for the column and the tubing preceding,
– mobile phase A: Water adjusted to pH 2.5 with a mixture of equal volumes of perchloric acid and water;
B: 0.03 per cent v/v solution of perchloric acid in acetonitrile ;
– flow rate: 1 ml per minute,
– spectrophotometer set at 215 nm,
– injection volume: 20 µl.
Time Mobile phase A Mobile phase B
(in min.) (per cent v/v) (per cent v/v)
0 – (5-t) 95 5
(5-t) – (60-t) 95-40 5-60
(60-t) – (65-t) 40-95 60-5
The isocratic step is described for a chromatographic system with a dwell volume (D) of 2 ml. If D is different from 2 ml, correct the gradient times with the value t. Calculated as, t = D-2
flow rate
Name Relative
retention time
perindopril 25
perindopril impurity B1 0.68
perindopril impurity K2 0.72
perindopril impurity E3 1.2
perindopril impurity F4 1.6
perindopril impurity H 5(may be eluted as 1 or 2 peaks) 1.8
1 (2S,3aS,7aS)-1-[(2S)-2-[[(1S)-1-carboxybutyl]amino]propanoyl]octahydro-1H-indole-2-carboxylic acid (perindoprilat),
2 (3S,5aS,9aS,10aS)-3-methyldecahydropyrazino[1,2-a]indole-1,4-dione,
3 (2S,3aS,7aS)-1-[(2S)-2-[[(1S)-1-[(1-methylethoxy)carbonyl]butyl]amino]propanoyl]octahydro-1H-indole-2-carboxylic acid,
4 ethyl (2S)-2-[(3S,5aS,9aS,10aS)-3-methyl-1,4-dioxodecahydropyrazino[1,2-a]indol-2(1H)-yl]pentanoate,
5 (2S,3aS,7aS)-1-[(2S)-2-[(5RS)-3-cyclohexyl-2-(cyclohexylimino)-4-oxo-5-propylimidazolidin-1-yl]propanoyl]octahydro-1H-indole-2-carboxylic acid,
Inject reference solutions (a). In the chromatogram obtained with reference solution (a) the peak to valley ratio between the peaks due to perindopril impurity B and perindoprilimpurity K is not less than 3.
Inject reference solution (b), reference solution (c) and the test solution. In the chromatogram obtained with the test solution, the area of peak due to perindopril impurity E is not more than 0.8 times the area of the principal peak in the chromatogram obtained with reference solution (b) (0.4 per cent), the area of peak due to perindopril impurity B is not more than 0.6 times the area of the principal peak in the chromatogram obtained with reference solution (b) (0.3 per cent), , the area of peak due to perindopril impurity F and perindopril impurity H for each impurity is not more than 0.4 times the area of the principal peak in the chromatogram obtained with reference solution (b) (0.2 per cent).The area of any other secondary peak is not more than 0.2 times the area of the principal peak in the chromatogram obtained with reference solution (b) (0.1 per cent) and the sum of the areas of all the secondary peaks is not more than twice the area of the principal peak in the chromatogram obtained with the reference solution (b) (1.0 per cent). Ignore any peak with an area of the principal peak in the chromatogram obtained with reference solution (c) (0.05 per cent).
Water (2.3.43). Not more than 1.0 per cent determined on 0.5 g.
Sulphated ash (2.3.18). Not more than 0.1 per cent.
Assay. Dissolve 0.16 g in 50 ml of anhydrous acetic acid. Titrate with 0.1 M perchloric acid, determining the end point potentiometrically (2.4.25). Carry out a blank titration.
1 ml of 0.1 M perchloric acid is equivalent to 0.02208 g of C23H43N3O5.
Storage. Store protected from moisture.