of Buprenorphine Hydrochlonde in Water for Injection Usual strength 300 ug per ml STANDARDSBuprenorphine Hydrochloride

a 0.01 % w/v solution in 0.01M hydrochlori acid exhibits a maximum at about 286 nm; absorbance at about 286 run. about 0. 33, Appendix 55.C Dissolve about 5 mg in 5 ml of hot water, add 2 ml of dilute hydrochloric acid and 2 ml of a 2% w/v solution of sodium nitrite and allow to stand for 10 minutes, a yellow colour is produced D: Dissolve 10 mg in 10 ml of hot water; add 2 ml of dilute nitric acid, shake and add 1 ml of silver nitrate solution; a white precipitate is produced. E: Melts at about 260* with decomposition, Appendix 8.8 Heavy metals Not more than 20 ppm, determined by the following method Moisten the residue obtained m the test for Sulphated ash with a few drops of hydrochloric acid, evaporate almost to dryness on a water-bath. Dissolve the residue in 10 ml of water by warming, cool, transfer to a test- tube with the aid of 10 ml of water, add 2 ml of dilute acetic acid The solution complies with the limt test for heavy metals, Method A, Appendix 3.12, using 5 ml of lead standard solution (2 ppm Pb) to prepare the standard solution Related substances Carry out the method for thin-layer chromatography, Appendix 4.6, using silica get GF254 as the coating substance and a mixture of 85 volumes of toluene, 15 volumes of methanol and 0.5 volume of strong ammonia solution as the mobile phase. Apply separately To the plate 5 ul of each of two freshly prepared solutions of the substance being examined in methanol containing (1) 0.5% w/v and (2) 0.025% w/v After removal of the plate, allow it lo dry in air and examine under ultra-violet light (254 nm) or expose lo iodine vapours. Any secondary spot in the chromatogram obtained with solution (1) is not more intense than the spot in chromatogram obtained with solution (2). Sulphated ash: Not more than 02%, Appendix 3.22. Loss on drying Not more than 1.0%, determined on 0.5 g by drying in an oven at 105° for 4 hours. Appendix 8.6 Assay Weigh accurately about 0.5 g, dissolve in 60 ml of anhydrous glacial acetic acid, add 10 nil of mercunic acetate solution and carry out Method B for non-aqueous titration, Appendix 3.45, using 0.1 ml of crystal violet solution as indicator to a green end-point. Perform a blank determination and make any necessary correction. Each ml of 0.1 M perchloric acid is equivalent to 0.05041 g of C29H41NO4HCI. BUPRENORPHINE HYDROCHLORIDE INJECTIONBuprenorphine InjectionBuprenorphine Hydrochloride Injection is a sterile solution. of Buprenorphine Hydrochlonde in Water for Injection Usual strength 300 ug per ml STANDARDSBuprenorphine Hydrochloride Injection contains not less than 90.0 per cent and not more than 110.0 per cent of the slated amount of buprenorphine hydrochlonde, C29H41NO4,HCL Identification. A Carry out the method for thin-layer chrornalography. Appendix 46, using silica gel HF254 as the coaling substance and a mixture of 85 volumes of toluene, 15 volumes of methonol and 0.5 volume of strong ammoniasolution as the mobile phase. Apply separately to the plate 10 ul of each of the following solutions For solution (1) transfer a volume of the injection. equivalent to 1 5 mg of Buprenorphine Hydrochlonde lo a 125-m1 separator, add 0.5 ml of dilute ammonia solution, shake and extract with three quantities, each of 10 ml, of chloroform, washing each chloroform extract with the same 10 ml of water and discard the water Evaporate the combined chloroform extracts to dryness on a water-bath and dissolve the residue in 1.5 ml of chloroform. For solution (2) -dissolve 1.5 mg of buprenorphine hydrochloride RS in 5 ml of 0.01 M hydrochloric acid, transfer the solution to a 125-ml separator and repeal the above procedure beginning at the
words "add 0.5 ml of dilute ammonia solution....." After removal of the plate, allow it to dry in a current of air and
examine under ultra-violet light (254 nm) or expose to iodine vapours.The principal spot in the chromatogram obtained with solution (1) corresponds to that in the chromatogram obtained with solution (2). B: To a volume equivalent to about 5 mg of Buprenorphine Hydrochloride in a 125-ml separator, add 1 ml of dilute ammonia solution and shake wrth three quantities, each of 10 ml, of chloroform Wash each chloroform extract with the same 10 ml of water and discard the washings. Evaporate the combined chloroform extracts to dryness on a water-bath and dissolve the residue in 50 ml of 0.1M hydrochloric acid. The light absorption of the resulting solution in the range 230 to 360 nm exhibits a maximum only ar about 286 run, Appendix 5.5 pH: Between 3.5 and b.5, Appendix 8.11. Other requirements Complies wrth the requirements of tests stated under Injectable Preparations (injections) Assay Measure accurately a volume equivalent to 1.5 rng of Buprenorphine Hydrochloride and transfer to a 25-m1 volumetric flask Add 1 ml of 1 M hydrochloric acid, 2 ml of a 2% w/v solution of sodium nitrite and shake well. Stopper the flask and allow to stand for 15 minutes. Dilute the solution lo volume with dilute ammonia solution and measure the absorbance of the resulting solution at the maximum at about 460 nm, Appendix 55. using as the blank a solution prepared in the same manner by treating 5 ml of waler instead of the preparation being examined Calculate the content of C29H41NO4,HCI from the absorbance obtained by repeating The procedure with 5 ml of a solution containing 300 ug of buprenorphine hydrochlonde RS per ml and from the declared content of