Amphotericin

General Notices

(Amphotericin B, Ph Eur monograph 1292) C47H73NO17__924__1397-89-3 Action and use Antifungal. Preparation Amphotericin Lozenges Amphotericin Oral Suspension Ph Eur DEFINITION Amphotericin B is a mixture of antifungal polyenes produced by the growth of certain strains of Streptomyces nodosus or by any other means. It consists mainly of amphotericin B which is (1R,3S,5R,6R,9R,11R,15S,16R,17R,18S,19E,21E,23E, 25E,27E,29E,31E,33R,35S,36R, 37S)-33-[(3-amino-3,6-dideoxy--D-mannopyranosyl)oxy]-1,3,5,6,9,11,17,37octahydroxy-15, 16,18-trimethyl-13-oxo-14,39-dioxabicyclo[33.3.1]nonatriaconta19,21,23,25,27,29,31heptaene-36-carboxylic acid. The potency is not less than 750 IU/mg, calculated with reference to the dried substance. Crown Copyright 2006 1 CHARACTERS A yellow or orange powder, practically insoluble in water, soluble in dimethyl sulphoxide and in propylene glycol, slightly soluble in dimethylformamide, very slightly soluble in methanol, practically insoluble in alcohol. It is sensitive to light in dilute solutions and is inactivated at low pH values. IDENTIFICATION _A. Dissolve 25 mg in 5 ml of dimethyl sulphoxide R and dilute to 50 ml with methanol R. Dilute 2 ml of the solution to 200 ml with methanol R. Examined between 300 nm and 450 nm (2.2.25), the solution shows 3 absorption maxima at 362 nm, 381 nm and 405 nm. The ratio of the absorbance measured at 362 nm to that measured at 381 nm is 0.57 to 0.61. The ratio of the absorbance measured at 381 nm to that measured at 405 nm is 0.87 to 0.93. _B. Examine by infrared absorption spectrophotometry (2.2.24), comparing with the spectrum obtained with amphotericin B CRS . If the spectra obtained show differences dry the substance to be examined at 60 C at a pressure not exceeding 0.7 kPa for 1 h and prepare a new spectrum.

_C. To 1 ml of a 0.5 g/l solution in dimethyl sulphoxide R, add 5 ml of phosphoric acid R to form a lower layer, avoiding mixing the 2 liquids. A blue ring is immediately produced at the junction of the liquids. Mix, an intense blue colour is produced. Add 15 ml of water R and mix; the solution becomes pale yellow. TESTS Content of tetraenes Not more than 10.0 per cent and not more than 5.0 per cent if intended for use in the manufacture of parenteral dosage forms, determined by the following method. Test solution_Dissolve 50.0 mg in 5 ml of dimethyl sulphoxide R and dilute to 50.0 ml with methanol R. Dilute 4.0 ml of the solution to 50.0 ml with methanol R. Reference solution (a)_Dissolve 50.0 mg of amphotericin B CRS in 5 ml of dimethyl sulphoxide R and dilute to 50.0 ml with methanol R. Dilute 4.0 ml of the solution to 50.0 ml with methanol R. Reference solution (b)_Dissolve 25.0 mg of nystatin CRS in 25 ml of dimethyl sulphoxide R and dilute to 250.0 ml with methanol R. Dilute 4.0 ml of the solution to 50.0 ml with methanol R. Measure the absorbances (2.2.25) of the test solution and of reference solutions (a) and (b) at the maxima at 282 nm and 304 nm respectively, using a 0.8 per cent V/V solution of dimethyl sulphoxide R in methanol R as the compensation liquid. Calculate the specific absorbance of the substance being examined, of nystatin CRS and of amphotericin B CRS at both wavelengths, each with reference to the dried substance, and calculate the percentage content of tetraenes using the following expression: Crown Copyright 2006 2 S1 and S2 = specific absorbances of the substance to be examined at 282 nm and 304 nm respectively, N1 and N2 = specific absorbance of nystatin CRS at 282 nm and 304 nm respectively, B1 and B2 = specific absorbance of amphotericin B CRS at 282 nm and 304 nm respectively, F = declared content of tetraenes in amphotericin B CRS. Loss on drying (2.2.32) Not more than 5.0 per cent, determined on 1.000 g by drying in an oven at 60 C at a pressure not exceeding 0.7 kPa. Sulphated ash (2.4.14) Not more than 3.0 per cent and not more than 0.5 per cent if intended for use in the manufacture of parenteral dosage forms, determined on 1.0 g. Bacterial endotoxins (2.6.14) Less than 1.0 IU/mg, if intended for use in the manufacture of parenteral dosage forms without

a further appropriate procedure for the removal of bacterial endotoxins. ASSAY Protect all solutions from light throughout the assay. Dissolve 25.0 mg in dimethyl sulphoxide R and dilute, with shaking, to 25.0 ml with the same solvent. Under constant stirring of this stock solution, dilute with dimethyl sulphoxide R to obtain solutions of appropriate concentrations (the following concentrations have been found suitable: 44.4, 66.7 and 100 IU/ ml). Final solutions are prepared by diluting 1 to 20 with 0.2 M phosphate buffer solution pH 10.5 so that they all contain 5 per cent V/V of dimethyl sulphoxide. Prepare the reference and the test solutions simultaneously. Carry out the microbiological assay of antibiotics (2.7.2). STORAGE Store protected from light, at a temperature of 2 C to 8 C. If the substance is sterile, store in a sterile, airtight, tamper-proof container. LABELLING The label states, where applicable, that the substance is suitable for use in the manufacture of parenteral dosage forms. Sumber : BP Vol. I & II

Amphotericin Oral Suspension

General Notices

Action and use Antifungal. DEFINITION Amphotericin Oral Suspension is a suspension of Amphotericin in a suitable flavoured vehicle. Amphotericin Oral Suspension should not be diluted. The oral suspension complies with the requirements stated under Oral Liquids and with the following requirements. IDENTIFICATION To a quantity containing the equivalent of 35 mg of amphotericin B add 10 ml of dimethyl sulphoxide , mix, add sufficient methanol to produce 250 ml and mix. If the solution is not clear, centrifuge a portion until clear. Dilute 2 ml of the clear supernatant liquid to 100 ml with methanol . The light absorption of the resulting solution, Appendix II B, in the range 300 to 450 nm exhibits three maxima, at 362, 381 and 405 nm. The ratio of the absorbance at the maximum at 362 nm to that at the maximum at 381 nm is 0.5 to 0.6. The ratio of the absorbance at the maximum at 381 nm to that at 405 nm is about 0.9. TESTS Content of tetraenes Complies with the test described under Amphotericin Lozenges but preparing solution (1) in the following manner. To a quantity containing the equivalent of 37.5 mg of amphotericin B add 5 ml of dimethyl sulphoxide and 25 ml of methanol and shake for 10 minutes, add sufficient methanol to produce 50 ml, mix thoroughly and filter. Dilute 4 ml of the filtrate to 50 ml with methanol . ASSAY To a weighed quantity containing the equivalent of 0.5 g of amphotericin B add 60 ml of dimethyl sulphoxide and shake for 20 minutes. Add sufficient dimethyl sulphoxide to produce 100 ml, mix, filter (Whatman No. 1 filter paper is suitable) and dilute 10 ml of the filtrate to 50 ml with dimethyl sulphoxide . Carry out the microbiological assay of antibiotics , Appendix XIV A. The precision of the assay is such that the fiducial limits of error are not less than 95% and not more than 105% of the estimated potency. Calculate the content of amphotericin B in the oral suspension taking each 1000 IU found to

be equivalent to 1 mg of amphotericin B. The upper fiducial limit of error is not less than Crown Copyright 2006 1 be equivalent to 1 mg of amphotericin B. The upper fiducial limit of error is not less than 90.0% and the lower fiducial limit of error is not more than 115.0% of the stated content. STORAGE Amphotericin Oral Suspension should be protected from light and used within the period stated on the label. LABELLING The quantity of active ingredient is stated in terms of the equivalent amount of amphotericin B. The label states the appropriate directions for using the preparation. Sumber : BP Vol. III