Definition
Alpha Tocopheryl Succinate Tablets contain RRR-Alpha-Tocopheryl Hydrogen Succinate. They are coated.
The tablets comply with the requirements stated under Tablets and with the following requirements.
Content of α-tocopherol, C29H50O2
95.0 to 105.0% of the stated amount.
Identification
(1) Shake a quantity of the powdered tablets containing the equivalent of 0.134 g of α-tocopherol with three 10 mL quantities of
ether, filter, evaporate the combined filtrates to 2 mL and then evaporate the solution to dryness under a current of nitrogen. Prepare a 0.5% w/v solution from a portion of the resulting residue in
cyclohexane.
(2) Dissolve 10 mg of the residue obtained in the preparation of solution (1) in 2 mL of 5
m ethanolic sulfuric acid, heat in a water bath for 1 minute, cool and add 2 mL each of
water and
cyclohexane. Shake for 1 minute, allow to separate and use the upper layer.
(3) 0.5% w/v of RRR
-α-tocopheryl succinate EPCRS in
cyclohexane.
(4) Prepare in the same manner as solution (2) but using 10 mg of RRR-α-tocopheryl succinate EPCRS in place of the substance being examined.
chromatographic conditions
(b) Use the mobile phase as described below.
(c) Apply 10 µL of each solution.
(d) Develop the plate to 15 cm.
mobile phase
20 volumes of ether and 80 volumes of cyclohexane.
system suitability
The test is not valid unless the chromatograms obtained with solutions (2) and (4) show two clearly separated spots.
confirmation
The principal spot in the chromatogram obtained with solution (1) is similar in position and size to that in the chromatogram obtained with solution (3).
Spray the plate with a mixture of 1 volume of hydrochloric acid, 4 volumes of a 0.25% w/v solution of iron (III) chloride hexahydrate in ethanol (96%) and 4 volumes of a 1% w/v solution of 1,10-phenanthroline hydrochloride in ethanol (96%). In the chromatograms obtained with solutions (2) and (4) the spot of higher Rf value, due to α-tocopherol, is orange.
B. In the Assay the principal peak in the chromatogram obtained with solution (1) shows a peak with the same retention time as the peak due to the methylated alpha tocopheryl succinate in the chromatogram obtained with solution (2).
Tests
Free tocopherol
Carry out the method for thin-layer chromatography, Appendix III A, using the following solutions.
(1) Shake a quantity of the powdered tablets containing the equivalent of 134 mg of α-tocopherol with three 10 mL quantities of
ether, filter and evaporate the combined filtrates to 2 mL, evaporate the final solution to dryness under a current of nitrogen and prepare a 0.5% w/v solution from the resulting residue in
cyclohexane.
chromatographic conditions
(b) Use the mobile phase as described below.
(c) Apply 10 µL of each solution.
(d) Develop the plate to 15 cm.
mobile phase
20 volumes of ether and 80 volumes of cyclohexane.
limits
Any secondary spot in the chromatogram obtained with solution (1) is not more intense than the principal spot in the chromatogram obtained with solution (2) (1%).
Assay
Carry out the method for gas chromatography, Appendix III B. Dissolve 0.15 g of dotriacontane (internal standard) in sufficient hexane to produce 100 mL (solution A).
(1) Mix a quantity of the powdered tablets containing the equivalent of 134 mg of α-tocopherol with 20 mL of
methanol, mix with the aid of ultrasound for 5 minutes and centrifuge for 15 minutes. To 4 mL of the clear supernatant liquid add 2 mL of
2,2-dimethoxypropane and 0.2 mL of
hydrochloric acid and allow to stand in the dark at room temperature for 1 hour. Evaporate to dryness on a water bath with the aid of a current of nitrogen and dissolve the residue in 10 mL of solution A.
(2) Dissolve 0.165 g of RRR
-α-tocopheryl succinate EPCRS in 20 mL of
methanol, add 2 mL of
2,2-dimethoxypropane and 0.2 mL of
hydrochloric acid and allow to stand in the dark at room temperature for 1 hour. Evaporate to dryness on a water bath with the aid of a current of nitrogen and dissolve the residue in 10 mL of solution A.
chromatographic conditions
(a) Use a borosilicate glass column (2 m × 4 mm) packed with
acid-washed, silanised diatomaceous support (100 to 120 mesh) (Chromosorb W/AW is suitable) coated with 2 to 5% of polymethylsiloxane.
(b) Use a rate of flow of carrier gas at values such that the required resolution is achieved (40 mL per minute is suitable).
(c) Set the temperature of the column such that the required resolution is achieved (a column temperature of 280° is suitable).
(d) Maintain the temperature of the injection port at 290° and the detector at 350°.
system suitability
The test is not valid unless, in the chromatogram obtained with solution (1), the retention times of dotriacontane and methyl α-tocopheryl succinate are about 8 minutes and 20 minutes, respectively.
determination of content
Calculate the content of C29H50O2 from the areas of the peaks due to dotriacontane and methyl α-tocopheryl succinate in the chromatograms obtained with solution (1) and solution (2) and from the declared content of C29H50O2 in RRR-α-tocopheryl succinate EPCRS.
Storage
Alpha Tocopheryl Succinate Tablets should be protected from light.
Labelling
The quantity of active ingredient is stated in terms of the equivalent amount of α-tocopherol.