Accelerated Degradation Experiment
Conditions
Procedure A: |
30 days exposure at 50 °C and 100% relative humidity. |
Procedure B: |
30 days exposure at 50 °C and 100%, relative humidity followed by 7 days at 70 °C and 100% relative humidity. |
Appearance of the substance
The substance, originally in the form of a white or almost white, crystalline powder or amorphous solid changes after procedure A into a liquid, then into a compact yellow mass turning to orange after procedure B.
Gain of moisture
Before exposure to heat and humidity: |
8.5% |
After procedure A: |
30% |
After procedure B: |
36% |
Chromatographic test
Adsorbent: Silica gel F-254
Solution applied to the plate: Substance dissolved in a mixture of chloroform-methanol (1:1).
Solvent system:
dichloromethane: ..............................77 volumes
methanol: ............................................8 volumes
ether: .................................................15 volumes
water: ...............................................1.2 volumes
Development conditions: The solvent is allowed to migrate until the front reaches a line 15 cm from the starting line.
Detection: The plate is examined under ultraviolet light at a wavelength of maximum output at about 254 nm, then left in a chromatographic chamber containing some iodine crystals until spots appear. A second plate is sprayed with a 5% solution of phosphomolybdic acid in acetic acid containing 5% of sulfuric acid.
Results: A reference solution of hydrocortisone sodium succinate shows a main spot and several weak secondary spots; the spot at Rf 0.18 corresponds to hydrocortisone, another at Rf 0.58. A solution of the sample subjected to procedure A shows that the spot at Rf 0.18 decreases, whereas the spot at Rf 0.58 increases. After procedure B, the change is even more significant.
Ultraviolet spectrophotometry
The absorbances of solutions of samples subjected to the test measured at about 241 nm and calculated with reference to the dried substance remain unchanged.
Colorimetric determination of the carbonyl-group in the A-ring
A solution in ethane 95% is treated with 4-aminophenazone in the presence of hydrochloric acid and measured at 380 nm.
No significant change is detected.
Colorimetric determination of the ketone chain
The product is treated with 4-nitrosodimethylaniline in alkaline medium. The N,N-dimethyl-p-phenylenediamine produced is detected with phenol in presence of ferricyanide ion and measured at 650 nm.
No significant change is detected.
Conclusion
Hydrocortisone sodium succinate is subject to decomposition under accelerated degradation conditions.
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