ammonia to 20 ml of a 0.05% w/v solution of atropine sulphate RS and complete the procedure described

lemperature and spray with dilute polassium indobismuthate solution. Any secondary spot in the chromatogram obtained with solution (1) is not more intense than the spot in the chromatogram obtained with solution (2) and not more than one such spot is more intense than the spot in the chromatogram obtained with solution (3). Sulphated ash: Not more than 0.2% Appendix 3.22: Water: Not more lhan 4,0% w/w, determined on 0.5 g, Appendix 3.24. Assay: Weigh accurately about 0.5 g, dissolve m 30 ml of anhydrous glacial acetic acid and carry out Method A for non-aqueous titration, Appendix 3.45, determining the end-point potenti ometrically.. Perform a blank determination and make any necessary correction: Each mlof 0.1M perchloric acid is equivalent to 0.06768 g of (C17H23NO3)2,H2S04,ATROPINE SULPHATE EYE OINTMENTAtropme Eye OintmentAtropine Sulphate Eye Ointment is a sterile preparation of Atropine Sulphate in an eye ointment base. Usual strength: 1.0% w/w. STANDARDSAtropine Sulphate Eye Ointment contains not less than 92.5 per cent and not more than 107.5 per cent of the stated amount of atropine sulphate, (C17H23N03)2,H204,H20. Identification: Carry out the method for thin-layer chromatography, Appendix 4.6, using silica gel G as the coating substance and a mixture of 50 volumes of chloroform, 40 volumes of acetone and 10 volumes of diethylamine as the mobile phase. Apply separately to the plale 5 µl of each of the following solutions. For solution. (1) dissolve a quantity of the ointment equivalent to 10 mg of Atropine Sulphate as completely as possible in 10 ml of light petroleum (boiling range 40° to 60°) and extract with two quantities, each of 10 ml, of 0.05M sulphuric acid, washing each acid solution with the same 5 ml of light petroleum (boiling range 40° to 60°) Mix Ihe acid solutions, make alkaline with dilute ammonia solution, and extract with two quantities, each of 15 ml, of chloroform. Remove the chloroform and dissolve the residue in 2 ml of ethanol (95%). Solution (2) contains 0.5% w/v of atropine sulphate RS in ethanol (95%). After removal of the plate, dry it at 105° for 20 minutes, allow it to cool lo room temperature and spray with polassium iodobismuthate solution. The principal spot in the chromatogram obtained with solution (1) corresponds to that in the chromalogram obtained with solution (2). Other requirements: Complies with the requirements of tests stated under Eye Ointments. Assay: Carry out Ihe method for gas chromatography, Appendix 4.2, using the following solutions. For solution (1) dissolve a quantity of Ihe eye ointment equivalent to 10 mg of Atropine Sulphate in 15 ml of chloroform, add 2 ml of a 0.5% w/v solution of homatropine hydrobromide RS (internal standard) in methanol (solulion A) and extract with two quantities, each of 10 ml, of 0.1M hydrochloric acid. Wash the combined exlracts with 10 ml of chloroform, add 2 ml of 5M ammonia. Extract with Iwo quantities, each of 10 ml, of chloroform, shake the combined extracts with 2 g of anhydrous sodium sulphate, filter and evaporate Ihe filtrate to dryness. Dissolve the residue m 5 ml of dichloromethane. To 1 ml of this solulion add 0.2 ml of a mixture of 4 volumes of N,O-bis(trimethylsilyl)acetamide and 1 volume of trimelhylchlorosilane, mix and allow to stand for 30 minutes. Prepare solution (2) in the same manner as solution (1) but omittmg the addition of solution A. For solution (3) add 2 ml of solulion A and 2 ml of 5M ammonia to 20 ml of a 0.05% w/v solution of atropine sulphate RS and complete the procedure described under solution (1) beginning at the words "Extract with two quantities, each of 10 ml, of chloroform......, ".The chromatographic procedure may be carried out using a glass column (1.5 m x 4 mm) packed with acid washed, silanised dialomaceous support (80 to 100 mesh) coated with 3% w/w of phenyl methyl silicone fluid (50% phenyl) (OV-17 is suitable) and maintained at 220° Calculate the content of (C17H23NO3)2,H2S04,H20 from Ihe declared content of (C17H23NO3)2,H2S04,H20 in atropine sulphate RS.ATROPINE SULPHATE INJECTION Atropine InieclionAtropine Sulphate Iniection is a slerile solution of Atropine Sulphate in Water for Iniection. Usual strenqths: