Dextran I for injection
`
`EUROPEAN PHARMACOPOEIA 5.0
`
`DEFINITION
`Dexpanthenol contains not less than 98.0 per cent and
`not more than the equivalent of 101.0 per cent of (2R)-2,
`4-dihydroxy-N-(3-hydroxypropyl)-3,3-dimethylbutanamide,
`calculated with reference to the anhydrous substance.
`
`CHARACTERS
`A colourless or slightly yellowish, viscous hygroscopic liquid,
`or a white or almost white, crystalline powder, very soluble
`in water, freely soluble in ethanol (96 per cent).
`
`IDENTIFICATION
`First identification: A, B.
`Second identification : A, C, D.
`A. It complies with the test for specific optical rotation (see
`Tests).
`B. Examine by infrared absorption spectrophotometry
`(2.2.24), comparing with the spectrum obtained with
`dexpanthenol CRS. Examine the substances using
`discs prepared as follows : dissolve the substance to be
`examined and the reference substance separately in
`1.0 ml of anhydrous ethanol R to obtain a concentration
`of 5 mg/ml. Place dropwise 0.5 ml of this solution on a
`disc of potassium bromide R. Dry the disc at 100-105 °C
`for 15 min.
`C. Examine the chromatograms obtained in the test for
`3-aminopropanol. The principal spot in the chromatogram
`obtained with test solution (b) is similar in position,
`colour and size to the principal spot in the chromatogram
`obtained with reference solution (a).
`D. To 1 ml of solution S (see Tests) add 1 ml of dilute sodium
`hydroxide solution Rand 0.1 ml of copper sulphate
`solution R. A blue colour develops.
`
`TESTS
`Solution S. Dissolve 2.500 g in carbon dioxide-free water R
`and dilute to 50.0 ml with the same solvent.
`Appearance of solution. Solution Sis clear (2.2.1) and not
`more intensely coloured than reference solution 8 6 (2.2.2,
`Method II).
`pH (2.2.3). The pH of solution S is not greater than 10.5.
`Specific optical rotation (2.2. 7). The specific optical rotation
`is + 29.0 to + 32.0, determined on solution S and calculated
`with reference to the anhydrous substance.
`3-Aminopropanol. Examine by thin-layer chromatography
`(2.2.27), using silica gel GR as the coating substance.
`Test solution (a). Dissolve 0.25 g of the substance to be
`examined in anhydrous ethanol R and dilute to 5 ml with
`the same solvent.
`Test solution (b). Dilute 1 ml of test solution (a) to 10 ml
`with anhydrous ethanol R.
`Reference solution (a). Dissolve the contents of a vial of
`dexpanthenol CRS in 1.0 ml of anhydrous ethanol R to
`obtain a concentration of 5 mg/ml.
`Reference solution (b). Dissolve 25 mg of 3-aminopropanol R
`in anhydrous ethanol Rand dilute to 100 ml with the same
`solvent.
`Apply separately to the plate 10 µl of each solution. Develop
`over a path of 15 cm using a mixture of 20 volumes of
`concentrated ammonia R, 25 volumes of methanol Rand
`55 volumes of butanol R. Allow the plate to dry in air,
`spray with a 100 g/1 solution of trichloroacetic acid R in
`methanol Rand heat at 150 °C for 10 min. Spray with a 1 g/)
`solution of ninhydrin R in methanol Rand heat at 120 °C
`until a colour appears. Any spot due to 3-aminopropanol
`
`in the chromatogram obtained with test solution (a) is not
`more intense than the spot in the chromatogram obtained
`with reference solution (b) (0.5 per cent).
`Heavy metals (2.4.8) . 12 ml of solution S complies with limit
`test A for heavy metals (20 ppm). Prepare the reference
`solution using lead standard solution (1 ppm Pb) R.
`Water (2.5.12). Not more than 1.0 per cent, determined on
`l.OOO g.
`Sulphated ash (2.4.14). Not more than 0.1 per cent,
`determined on 1.0 g.
`
`ASSAY
`To 0.400 g add 50.0 ml of 0.1 M perchloric acid. Boil under
`a reflux condenser for 5 h protected from humidity. Allow
`to cool. Add 50 ml of dioxan R by rinsing the condenser,
`protected from humidity. Add 0.2 ml of naphtholbenzein
`solution Rand titrate with 0.1 M potassium hydrogen
`phthalate until the colour changes from green to yellow.
`Carry out a blank titration.
`1 ml of 0.1 M perchloric acid is equivalent to 20.53 mg
`of C9H19N04•
`
`STORAGE
`In an airtight container.
`
`Ol/2005:1506
`
`DEXTRAN 1 FOR INJECTION
`
`Dextranum 1 ad iniectabile
`DEFINITION
`Dextran 1 for injection is a low molecular weight fraction of
`dextran, consisting of a mixture of isomaltooligosaccharides.
`The average relative molecular mass is about 1000.
`
`PRODUCTION
`It is obtained by hydrolysis and fractionation of dextrans
`produced by fermentation of sucrose using Leuconostoc
`mesenteroides strain NRRL 8-512 = CIP 78.59 or
`substrains thereof (for example L. mesenteroides B-512
`F = NCTC 10817).
`It is prepared in conditions designed to minimise the risk
`of microbial contamination.
`
`CHARACTERS
`A white or almost white powder, hygroscopic, very soluble in
`water, very slightly soluble in alcohol.
`
`IDENTIFICATION
`A. Dissolve 3.000 g in water R, heat on a water-bath and
`dilute to 100.0 ml with the same solvent. The specific
`optical rotation (2.2.7) is+ 148 to+ 164, calculated with
`reference to the dried substance. Dry an aliquot of the
`solution first on a water-bath and then to constant weight
`in vacuo at 70 °C. Calculate the dextran content after
`correction for the content of sodium chloride.
`B. Examine by infrared absorption spectrophotometry
`(2.2.24), comparing with the spectrum obtained with
`dextran 1 CRS. Prepare the discs as follows: to 1-2 mg
`add one or a few drops of water R; grind in an agate
`mortar for 1-2 min; add about 300 mg of potassium
`bromide Rand mix to a slurry (do not grind); dry in vacuo
`at 40 °C for 15 min, crush the residue (if it is not dry,
`dry for another 15 min). Prepare a disc using potassium
`bromide R. Run the infrared spectrum with a blank disc
`using potassium bromide R in the reference beam.
`
`1408
`
`See the information section on general monographs (cover pages)
`
`PGR2020-00009
`Pharmacosmos A/S v. American Regent, Inc.
`Petitioner Ex. 1038 - Page 1
`
`

`

`EUROPEAN PHARMACOPOEIA 5.0
`
`Dextran 40 for injection
`
`C. It complies with the test for molecular-mass distribution
`(see Tests).
`
`TESTS
`Solution S. Dissolve 7.5 g in carbon dioxide-free water R,
`heat on a water-bath and dilute to 50 ml with the same
`solvent.
`Absorbance (2.2.25). Measure the absorbance of solution S
`at 375 nm. The absorbance is not more than 0.12.
`Acidity or alkalinity. To 10 ml of solution S add 0.1 ml of
`phenolphthalein solution R. The solution is colourless.
`Add 0.2 ml of 0.01 M sodium hydroxide. The solution is
`pink. Add 0.4 ml of 0.01 M hydrochloric acid. The solution
`is colourless. Add 0.1 ml of methyl red solution R. The
`solution is red or orange.
`Nitrogen-containing substances. Carry out the
`determination of nitrogen by sulphuric acid digestion (2.5.9),
`using 0.200 g and heating for 2 h. Collect the distillate in a
`mixture of 0.5 ml of bromocresol green solution R, 0.5 ml
`of methyl red solution Rand 20 ml of water R. Titrate with
`0.01 M hydrochloric acid. Not more than 0.15 ml of 0.01 M
`hydrochloric acid is required to change the colour of the
`indicator (llO ppm N).
`Sodium chloride. Not more than 1.5 per cent. Accurately
`weigh 3-5 g and dissolve in 100 ml of water R. Add 0.3 ml
`of potassium chromate solution Rand titrate with 0.1 M
`silver nitrate until the yellowish-white colour changes to
`reddish-brown.
`1 ml of 0.1 M silver nitrate is equivalent to 5.844 mg of NaCl.
`Molecular-mass distribution. The average molecular mass
`(MJ is 850 to ll50. The fraction with less than 3 units of
`glucose is less than 15 per cent, the fraction with more than
`9 units of glucose is less than 20 per cent.
`
`Examine by size-exclusion chromatography (2.2.30).
`Test solution. Dissolve 6.0-6.5 mg of the substance to be
`examined in 1.0 ml of the mobile phase.
`Reference solution (a). Dissolve 6.0-6.5 mg of dextran 1 CRS
`in 1.0 ml of the mobile phase.
`Reference solution (b). Dissolve the content of an ampoule
`of isomaltooligosaccharide CRS in 1 ml of the mobile
`phase, and mix. This corresponds to approximately 45 µg
`of isomaltotriose (3 glucose units), approximately 45 µg of
`isomaltononaose (9 glucose units), and approximately 60 µg
`of sodium chloride per 100 µI.
`
`The chromatographic procedure may be carried out using:
`
`2 columns, 30 cm long and 10 mm in internal diameter,
`in series, prepacked with a packing material of dextran
`covalently bound to highly cross-linked porous agarose
`beads, allowing resolution of oligosaccharides in
`the molecular mass range of 180 to 3000, kept at a
`temperature of 20-25 °C,
`
`as mobile phase at a flow rate of 0.07-0.08 ml/min
`maintained constant to ± 1 per cent, a 2.92 g/1 solution
`of sodium chloride R,
`as detector a differential refractometer.
`
`Inject 100 µl of reference solution (b) and record
`the chromatogram for definition of the positions of
`isomaltotriose, isomaltononaose and sodium chloride. Inject
`100 µl of the test solution and 100 µl of reference solution (a)
`and record the chromatograms. Determine the peak areas.
`Disregard any peak due to sodium chloride.
`
`Calculate the average relative molecular mass Mw and the
`amount of the fraction with less than 3 and more than 9
`glucose units, of dextran 1 CRS and of the substance to be
`examined. The test is not valid unless the values obtained
`for dextran 1 CRS are within the values stated on the label.
`Mw = LWi X m ;
`
`Mw
`average molecular mass of the dextran,
`m,
`molecular mass of oligosaccharide i,
`w,
`weight proportion of oligosaccharide i.
`Use the following molecular mass values for the calculation:
`
`m,
`180
`342
`504
`666
`828
`990
`ll52
`1314
`1476
`1638
`1800
`1962
`2124
`2286
`2448
`2610
`2772
`2934
`3096
`
`Oligosaccharide i
`glucose
`isomaltose
`isomaltotriose
`isomaltotetraose
`isomaltopentaose
`isomaltohexaose
`isomaltoheptaose
`isomaltooctaose
`isomaltononaose
`isomaltodecaose
`isomaltoundecaose
`isomaltododecaose
`isomaltotridecaose
`isomaltotetradecaose
`isomaltopentadecaose
`isomaltohexadecaose
`isomaltoheptadecaose
`isomaltooctadecaose
`isomaltononadecaose
`Heavy metals (2.4.8). Dilute 20 ml of solution S to 30 ml
`with water R. 12 ml of this solution complies with limit
`test A (10 ppm). Prepare the reference solution using lead
`standard solution (1 ppm Pb) R.
`Loss on drying (2.2.32). Not more than 5.0 per cent,
`determined on 5.000 g by drying in an oven at 100-105 °C
`for 5 h.
`Bacterial endotoxins (2.6.14) : less than 25 IU/ g.
`Microbial contamination. Total viable aerobic count (2. 6.12)
`not more than 102 micro-organisms per gram, determined
`by plate-count. It complies with the test for Escherichia
`coli (2.6.13).
`
`01/2005:0999
`
`DEXTRAN 40 FOR INJECTION
`
`Dextranum 40 ad iniectabile
`DEFINITION
`Dextran 40 for injection is a mixture of polysaccharides,
`principally of the a-1,6-glucan type.
`The average relative molecular mass is about 40 000.
`
`General Notices (1) apply to all monographs and other texts
`
`1409
`
`PGR2020-00009
`Pharmacosmos A/S v. American Regent, Inc.
`Petitioner Ex. 1038 - Page 2
`
`