Product : Ethinyl Estradiol IP

Analysis As Per

IP

Specification No.

FP-018

Page No.

1 of 2

Sr. No.

Test

Specification

1.              

Description

White or slightly yellowish white, crystalline powder.

2.              

Solubility

Freely soluble in ethanol (95%) and in ether; Sparingly soluble in chloroform; practically insoluble in water. It dissolves in dilute solutions of alkalis.

3.              

Identification

a)      The infrared absorption spectrum is concordant with the reference spectrum of Levonorgestrel or with the spectrum obtained from Levonorgestrel RS.

b)      In the test for related substances, the principal spot in the chromatogram obtained with solution (2) corresponds to that in the chromatogram obtained with solution (4)

c)      Dissolve about 1 mg in 1 ml of sulphuric acid; an orange-red colour develops which exhibits a greenish fluorescence when examined under ultra-violet light (365 nm). Add the solution to 10 ml of water; the colour changes to violet and a violet precipitates is produced

d)      Melts between 181 ° C and 185 ° C

4.              

Clarity and colour of solution

A 5.0 % w/v solution in ethanol is clear (appendix 6.1 of IP) and not more intensely coloured than reference solution BYS6 (appendix 6.2 of IP)

5.              

Light absorption

Abosrbance of a 0.01 % w/v solution in ethanol (95 %) at about 281 nm, 0.69 to 0.73

6.              

Specific Optical Rotation

-27.0º to –30.0º determined at 20 ° in a 5 % w/v solution in pyridine

7.              

Sulphated Ash

Not more than 0.3 %

8.              

Loss on Drying

Not more than 1.0 %

9.              

Assay

(By HPLC)

97.0 % to 102 % calculated on dry basis

Dissolve 0.2 gm in 40 ml of tetrahydrofuran, add 5 ml of a 10% w/v solution of silver nitrate and titrate with 0.1 M sodium hydroxide, determining the end point potentiometrically. Perform a blank determination and make any necessary correction. Each ml 0f 0.1 M sodium hydroxide is equivalent to 0.02964 gm of  C₂₀H₂₄O₂

10.          

Related Substances

(By TLC)

Carry out the method for thin layer chromatography using silica gel G as the coating substance and a mixture of 90 volumes of toluene and 10 volumes of ethanol (95%) as the mobile phase. Apply separately to the plate 5 ml of each of five solutions in a mixture of 90 volumes of chloroform and 10 volumes of methanol. Solutions (1) 2.0 % w/v, (2) 0.10% w/v and (3) 0.020 % w/v of the substances being examined. Solution (4) contains 0.1 % w/v of ethinylestardiol RS. Solution (5) contains 0.02 % w/v of estrone RS. After removal of the plate, allow it to dry in air until odour of the solvent is no longer detectable, heat at 110 °C for 10 minutes and spray the hot plate with ethanolic sulphuric acid (20% v/v). Heat again at 110 °C for 10 min. and examine under ultra violet light (365 nm). In the chromatogram obtained with solutiom (1) any spot corresponding to estrone is not more intense than the spot in chromatogram obtained with solution (5) and any other secondary spot is not more intense than the spot in the chromatogram obtained with solution (3).