EUROPEAN PHARMACOPOEIA 5.

Xanthan gum

01/2005:1277 Test solution. To 200 ml of water R in a 1000 ml round-bottomed flask, add 5.0 g of the substance to be examined and 1 ml of a 10 g/l emulsion of dimeticone R in XANTHAN GUM liquid paraffin R, stopper the flask and shake for 1 h. Distil about 90.0 ml, mix the distillate with 4.0 ml of the internal Xanthani gummi standard solution and dilute to 100.0 ml with water R. DEFINITION Reference solution. Dilute a suitable quantity of 2-propanol R, accurately weighed, with water R to obtain Xanthan gum is a high-molecular-mass anionic a solution having a known concentration of 2-propanol of polysaccharide produced by fermentation of carbohydrate about 1 mg/ml. To 4.0 ml of this solution add 4.0 ml of with Xanthomonas campestris. It consists of a principal the internal standard solution and dilute to 100.0 ml with chain of (14)-linked D-glucose units with trisaccharide water R. side chains, on alternating anhydroglucose units, consisting of a glucuronic acid unit included between two The chromatographic procedure may be carried out using : mannose units. Most of the terminal units contain a pyruvate a column 1.8 m long and 4.0 mm in internal diameter moiety and the mannose unit adjacent to the principal chain packed with styrene-divinylbenzene copolymer R, may be acetylated at C-6. helium for chromatography R as the carrier gas at a flow Xanthan gum has a molecular mass of approximately 1 106. rate of 30 ml/min, It contains not less than 1.5 per cent of pyruvoyl groups a flame-ionisation detector, (C3H3O2 ; relative mass of the group 71.1), calculated with reference to the dried substance. Xanthan gum exists as the maintaining the temperature of the column at 165 C, and that of the injection port and of the detector at 200 C. sodium, potassium or calcium salt. Inject 5 l of the test solution and 5 l of the reference CHARACTERS solution. The retention time of 2-methyl-2-propanol is about 1.5 relative to that of 2-propanol. A white or yellowish-white, free-flowing powder, soluble in water giving a highly viscous solution, practically insoluble Other polysaccharides. Examine by thin-layer in organic solvents. chromatography (2.2.27), using a TLC silica gel plate R. Test solution. To 10 mg in a thick-walled centrifuge test tube IDENTIFICATION add 2 ml of a 230 g/l solution of trifluoroacetic acid R, A. In a flask, suspend 1 g in 15 ml of 0.1 M hydrochloric shake vigorously to dissolve the forming gel, stopper the test acid. Close the flask with a fermentation bulb containing barium hydroxide solution R and heat carefully for 5 min. tube, and heat the mixture at 120 C for 1 h. Centrifuge the hydrolysate, transfer the clear supernatant liquid carefully The barium hydroxide solution shows a white turbidity. into a 50 ml flask, add 10 ml of water R and evaporate the B. To 300 ml of water R, previously heated to 80 C and solution to dryness under reduced pressure. Take up the stirred rapidly with a mechanical stirrer in a 400 ml residue thus obtained in 10 ml of water R and evaporate to beaker, add, at the point of maximum agitation, a dry dryness under reduced pressure. Wash three times with blend of 1.5 g of carob bean gum R and 1.5 g of the 20 ml of methanol R and evaporate under reduced pressure. substance to be examined. Stir until the mixture forms To the resulting clear film which has no odour of acetic acid, a solution, and then continue stirring for 30 min or add 0.1 ml of water R and 1 ml of methanol R. Centrifuge to longer. Do not allow the water temperature to drop below separate the amorphous precipitate. Dilute the supernatant 60 C during stirring. Discontinue stirring and allow the liquid, if necessary, to 1 ml with methanol R. mixture to stand for at least 2 h. A firm rubbery gel forms Reference solution. Dissolve 10 mg of glucose R and 10 mg after the temperature drops below 40 C but no such of mannose R in 2 ml of water R and dilute to 10 ml with gel forms in a 1 per cent control solution of the sample prepared in the same manner but omitting the carob bean methanol R. Apply separately to the plate, as bands, 5 l of each solution. gum. Develop over a path of 15 cm using a mixture of 10 volumes TESTS of a 16 g/l solution of sodium dihydrogen phosphate R, 40 volumes of butanol R and 50 volumes of acetone R. pH (2.2.3). The pH of a 10.0 g/l solution is 6.0 to 8.0. Spray evenly with a solution of 0.5 g of diphenylamine R in Viscosity (2.2.10). The viscosity at 24 1 C is not less 25 ml of methanol R to which 0.5 ml of aniline R and 2.5 ml than 600 mPas. Add 3.0 g within 45 s to 90 s into 250 ml of phosphoric acid R have been added. Heat for 5 min at of a 12 g/l solution of potassium chloride R in a 500 ml 120 C. Examine in daylight. The test is not valid unless the beaker stirring with a low-pitch propeller-type stirrer rotating chromatogram obtained with the reference solution shows at 800 r/min. When adding the substance take care that two clearly separated greyish-brown zones due to glucose agglomerates are destroyed. Add an additional quantity of and mannose in the middle third. The chromatogram 44 ml of water R, to rinse any adhering residue from the obtained with the test solution shows corresponding zones. walls of the beaker. Stir the preparation at 800 r/min for 2 h In addition, one weak reddish and two faint bluish-grey whilst maintaining the temperature at 24 1 C. Determine bands may be visible just above the starting line. One or two the viscosity within 15 min using a rotating viscosimeter set bluish-grey bands may also be seen in the upper quarter of at 60 r/min and equipped with a rotating spindle 12.7 mm the chromatogram. No other bands are visible. in diameter and 1.6 mm high which is attached to a shaft Loss on drying (2.2.32). Not more than 15.0 per cent, 3.2 mm in diameter. The distance from the top of the cylinder to the lower tip of the shaft being 25.4 mm, and the determined on 1.000 g by drying in an oven at 100 C to 105 C for 2.5 h. immersion depth being 50.0 mm. Total ash (2.4.16) : 6.5 per cent to 16.0 per cent. 2-Propanol. Not more than 750 ppm, determined by gas chromatography (2.2.28) using 2-methyl-2-propanol R as Microbial contamination. Total viable aerobic count the internal standard. (2.6.12) not more than 103 bacteria and 102 fungi per gram, Internal standard solution. Dilute 0.50 g of determined by plate count. It complies with the test for 2-methyl-2-propanol R to 500 ml with water R. Escherichia coli (2.6.13). General Notices (1) apply to all monographs and other texts 2715

Xylazine hydrochloride for veterinary use

EUROPEAN PHARMACOPOEIA 5.0

ASSAY Test solution. Dissolve a quantity of the substance to be examined corresponding to 120.0 mg of the dried substance in water R and dilute to 20.0 ml with the same solvent. Reference solution. Dissolve 45.0 mg of pyruvic acid R in water R and dilute to 500.0 ml with the same solvent. Place 10.0 ml of the test solution in a 50 ml round-bottomed flask, add 20.0 ml of 0.1 M hydrochloric acid and weigh. Boil on a water-bath under a reflux condenser for 3 h. Weigh and adjust to the initial mass with water R. In a separating funnel mix 2.0 ml of the solution with 1.0 ml of dinitrophenylhydrazine-hydrochloric solution R. Allow to stand for 5 min and add 5.0 ml of ethyl acetate R. Shake and allow the solids to settle. Collect the upper layer and shake with three quantities, each of 5.0 ml, of sodium carbonate solution R. Combine the aqueous layers and dilute to 50.0 ml with sodium carbonate solution R. Mix. Treat 10.0 ml of the reference solution at the same time and in the same manner as for the test solution. Immediately measure the absorbance of the two solutions (2.2.25) at 375 nm, using sodium carbonate solution R as the compensation liquid. The absorbance of the test solution is not less than that of the reference solution, which corresponds to a content of pyruvic acid of not less than 1.5 per cent.

pH (2.2.3) : 4.0 to 5.5 for solution S. Impurity A : maximum 100 ppm. Solution A. Dissolve 0.25 g of the substance to be examined in methanol R and dilute to 10 ml with the same solvent. This solution is used to prepare the test solution. Solution B. Dissolve 50 mg of 2,6-dimethylaniline R in methanol R and dilute to 100 ml with the same solvent. Dilute 1 ml of the solution to 100 ml with methanol R. This solution is used to prepare the reference solution. Using 2 flat-bottomed tubes with an inner diameter of about 10 mm, place in the first tube 2 ml of solution A, and in the second tube 1 ml of solution B and 1 ml of methanol R. To each tube add 1 ml of a freshly prepared 10 g/l solution of dimethylaminobenzaldehyde R in methanol R and 2 ml of glacial acetic acid R and allow to stand at room temperature for 10 min. Compare the colours in diffused daylight, viewing vertically against a white background. Any yellow colour in the test solution is not more intense than that in the reference solution. Related substances. Liquid chromatography (2.2.29). Prepare the solutions immediately before use. Solvent mixture. Mix 8 volumes of acetonitrile R, 30 volumes of methanol R and 62 volumes of a 2.72 g/l solution of potassium dihydrogen phosphate R adjusted to pH 7.2 with dilute sodium hydroxide solution R. 01/2005:1481 Test solution. Dissolve 0.100 g of the substance to be examined in the solvent mixture and dilute to 20.0 ml with XYLAZINE HYDROCHLORIDE the solvent mixture. Reference solution. Dissolve 5.0 mg of the substance to FOR VETERINARY USE be examined, 5.0 mg of 2,6-dimethylaniline R, 5.0 mg of xylazine impurity C CRS and 5.0 mg of xylazine Xylazini hydrochloridum ad usum impurity E CRS in acetonitrile R and dilute to 100.0 ml with veterinarium the same solvent. Dilute 1.0 ml of this solution to 10.0 ml with the solvent mixture. Column : size : l = 0.15 m, = 3.9 mm, stationary phase : end-capped octylsilyl silica gel for chromatography with polar incorporated groups R (5 m), C12H17ClN2S Mr 256.8 temperature : 40 C. DEFINITION Mobile phase : N-(2,6-Dimethylphenyl)-5,6-dihydro-4H-1,3-thiazin-2-amine mobile phase A : mix 30 volumes of methanol R hydrochloride. and 70 volumes of a 2.72 g/l solution of potassium Content : 98.0 per cent to 102.0 per cent (dried substance). dihydrogen phosphate R adjusted to pH 7.2 with dilute sodium hydroxide solution R, CHARACTERS mobile phase B : methanol R, acetonitrile R (30:70 V/V), Appearance : white or almost white, crystalline powder, hygroscopic. Time Mobile phase A Mobile phase B Solubility : freely soluble in water, very soluble in methanol, (min) (per cent V/V) (per cent V/V) freely soluble in methylene chloride.
0 - 15 89 28 28 28 89 89 11 72 72 72 11 11

IDENTIFICATION A. Infrared absorption spectrophotometry (2.2.24). Preparation : discs. Comparison : xylazine hydrochloride CRS. B. It gives reaction (b) of chlorides (2.3.1). TESTS Solution S. Dissolve 5.0 g in carbon dioxide-free water R prepared from distilled water R, heating at 60 C if necessary ; allow to cool and dilute to 50.0 ml with the same solvent. Appearance of solution. Solution S is not more opalescent than reference suspension II (2.2.1) and is colourless (2.2.2, Method II). 2716

15 - 21 21 - 22 22 - 33

Flow rate : 1.0 ml/min. Detection : spectrophotometer at 230 nm. Equilibration : for at least 30 min with a mixture of 28 volumes of mobile phase A and 72 volumes of mobile phase B. Injection : 20 l. Relative retention with reference to xylazine (retention time = about 7.5 min) : impurity A = about 0.8 ; impurity E = about 1.6 ; impurity C = about 2.2.

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