The chapter reprint attached with this page is a true copy of the following:
`
`USP28-NF23, Official Monographs/Dextran, pp 601-602
`
`© 2004 The United States Pharmacopeial Convention, Inc.
`
`
`
`
`
`
`
`
`
`
`IPR2019-01142
`Pharamacosmos v. American Regent
`Petitioner Ex. 1007 - Page 1
`
`

`

`USP28
`
`Official Monographs I Dextran
`
`601
`
`ionization detector and contains a suitable column, 1.8 m x 2.0 mm,
`packed with 5% liquid phase G2 on support SIA. The column and
`injection port are maintained isothermally at 170° and 180°,
`respectively. Using a suitable carrier gas, adjust the flow rate so
`that the derivatized pantolactone elutes in about 4 minutes.
`Chromatograph five injections of the Standard solution 2, and
`record the peak responses as directed under Procedure: the relative
`standard deviation of the peak response ratios (Rs) of the five
`injections is not more than 2.0%. The retention time of the derivatized
`pantolactone is about 0. 75 relative to that of the derivatized internal
`standard. In a suitable chromatogram, the resolution factor between
`the two peaks is not less than 2.0.
`Procedure- Inject about 0.5 11L of Standard solution 2 into the gas
`chromatograph, record the chromatogram to obtain not less than 40%
`of maximum recorder response, and measure the peak responses of
`the derivatized pantolactone and the derivatized internal standard.
`Similarly, inject about 0.5 11L of the Test solution, record the
`chromatogram, and measure the peak responses of the corresponding
`components. Calculate the quantity, in mg, of pantolactone in the
`portion of Preparation taken by the formula:
`
`0.4Cs(Rul Rs),
`
`in which Cs is the concentration, in mg per mL, ofUSP Pantolactone
`RS in Standard solution 1, and Ru and Rs are the ratios of the peak
`response due to the pantolactone to that due to the internal standard
`obtained from the Test solution and the Standard solution 2,
`respectively.
`Other requirements-It meets the requirements for Refractive
`index, Water, Residue on ignition, Limit of aminopropanol, and Assay
`under Dexpanthenol.
`
`Dextran 1
`
`» Dextran 1 is a low molecular weight fraction of
`dextran, consisting of a mixture of isomaltooligosac(cid:173)
`charides. It is obtained by controlled hydrolysis and
`fractionation of dextrans produced by fermentation of
`Leuconostoc mesenteroides (strain NRRL B-512; CIP
`78.59, or its sub-strains, for example L. mesenteroides B-
`512F; NCTC, 10817), in the presence of sucrose. It is a
`glucose polymer in which the linkages between glucose
`units are almost exclusively r:x-1 ,6. Its weight-average
`molecular weight is about 1000.
`
`Packaging and storage--Store in well-closed containers at a
`temperature between 4° and 30°.
`Labeling-Where it is intended for use in preparing injectable
`dosage forms, the label states that it is sterile or must be subjected to
`further processing during the preparation of injectable dosage forms.
`USP Reference standards ( 11 )-USP Dextran 1 RS. USP
`Endotoxin RS.
`Identification-
`A:
`Infrared Absorption (197K)-To I to 2 mg each of USP
`Dextran 1 RS and the sample add one to two drops of water, grind in
`an agate mortar for I to 2 minutes, add about 300 mg of potassium
`bromide, and mix to a slurry. [NOTE- Do not grind.] Dry under
`vacuum at 40° for 15 minutes, and if it is not dry, continue drying for
`another 15 minutes. Crush the residue, prepare a disk, and run the IR
`spectrum with a blank potassium bromide disk in the reference beam.
`It meets the requirements of the test for Molecular weight
`B:
`distribution and average molecular weight.
`Absorbance (851)-The absorbance of a 15% solution in water at
`375 nm is not more than 0.12, water being used as the blank.
`between + 148° and + 164 o at 20°, for a
`Specific rotation (781 S):
`solution in water, on the dried basis (dry at 70° under vacuum to
`constant weight), and corrected for the content of sodium chloride.
`Microbial limits (61)-The total aerobic microbial count does not
`exceed 102 cfu per g, determined by plate-count; and the total
`combined molds and yeasts count does not exceed 10 cfu per g.
`
`Bacterial endotoxins (85) (where it is labeled as intended for use in
`the preparation of injectables):
`not more than 25.0 USP Endotoxin
`Units per g.
`between 4.5 and 7.0, in a 15% solution in water.
`pH (791):
`Loss on drying (731 )-Dry it at 100° to 105° for 5 hours: it loses not
`more than 5.0% of its weight.
`not more than 5 11g per g.
`Heavy metals, Method II (231):
`Limit of alcohol and related impurities-
`Test solution-Proceed as directed for Test solution in the test for
`Limit of alcohol and related impurities under Dextran 40, except to
`use 5.0 g of Dextran I.
`Standard solution, Chromatographic system, and Procedure(cid:173)
`Proceed as directed in the test for Limit of alcohol and related
`impurities under Dextran 40. The total area of peaks from impurities
`in the Test solution does not exceed the area of the n-propyl alcohol
`solution peak.
`Limit of sodium chloride--Dissolve 5 g of Dextran 1, accurately
`weighed, in 100 mL of water. Add 0.2 mL of potassium chromate TS,
`and titrate with 0.1 N silver nitrate VS (see Titrimet1y (541 )). Each
`mL of 0.1 N silver nitrate is equivalent to 5.844 mg of sodium
`chloride: not more than 1.5% of sodium chloride is found.
`Limit of nitrogenous impurities (461 ) (where it is labeled as
`intended for use in the preparation of injectables)-
`Sulfate solution-To I 000 mL sulfuric acid add 5 g of anhydrous
`cupric sulfate and 500 g of potassium sulfate. Dissolve by heating,
`and store at 60°. [NOTE- If storage at 60° is not possible, prepare a
`smaller quantity of Sulfate solution on the day of use, adjusting
`proportions accordingly.]
`Indicator-Dilute a mixture of 20 mL of a 0.1% solution of
`bromocresol green in alcohol and 4 mL of methyl red TS with water
`to 100 mL.
`Procedure- Transfer 0.2 g Dextran 1, accurately weighed, to a
`micro-Kjeldahl flask. Add 4 mL of Sulfate solution. Heat until the
`solution exhibits a clear green color and the sides of the flask are free
`from carbonaceous material. Cool, cautiously add 30 mL of water,
`mix, and transfer the solution to a steam distillation unit. Rinse the
`Kjeldahl flask with three 5-mL portions of water, adding the washings
`to the solution. Add 15 mL of 45% sodium hydroxide solution,
`immediately close the distillation apparatus, and start steam
`distillation immediately. Receive the distillate in 1 mL of Indicator
`and sufficient water to cover the end of the condensing tube. Upon
`completion of the distillation, remove the receiving flask, and rinse
`the end of the condensing tube with a small quantity of water, adding
`the rinse to the distillate. Titrate the distillate with 0.0 I 0 N
`hydrochloric acid until the color changes from blue to reddish
`violet. Perform a blank determination, and make any necessary
`correction. The corrected volume of 0.0 I 0 N hydrochloric acid
`required to change the color does not exceed 0.15 mL (I I 0 ppm of
`nitrogen).
`Molecular weight distribution and average molecular weight(cid:173)
`Mobile phase-Prepare a filtered and degassed solution of sodium
`chloride containing 2.9 g per liter.
`Calibration solution-Prepare a solution containing about 0.45 mg
`of isomaltotriose (3 glucose units) and 0.60 mg of sodium chloride
`per mL.
`Reference solution-Prepare a solution of USP Dextran I RS in
`Mobile phase containing 6.0 to 6.5 mg per mL.
`Test solution-Prepare a solution of Dextran I in Mobile phase
`containing 6.0 to 6.5 mg per mL.
`Chromatographic system (see Chromatography (62 1))-The
`liquid chromatograph is equipped with a differential refractive
`index detector and two 10-mm x 30-cm columns in series that
`contain packing L54 and are maintained at 20- 25°. The flow rate is
`0.07 to 0.08 mL per minute, maintained constant to ± I%.
`Procedure-Inject about I 00 11L of the Calibration solution,
`record the chromatogram, and note the retention times of the peaks.
`Separately inject equal volumes (about 100 IlL) of the Reference
`solution and Test solution, and record the chromatograms. Using the
`retention times in the chromatogram of Calibration solution, identify
`the peaks due to isomaltotriose and sodium chloride in the
`chromatograms of Reference solution and Test solution. Disregard
`the peak due to sodium chloride in Reference solution and Test
`
`
`
`
`IPR2019-01142
`Pharamacosmos v. American Regent
`Petitioner Ex. 1007 - Page 2
`
`

`

`602
`
`Dextran
`
`/ Official Monographs
`
`USP28
`
`solution. Calculate the weight-average molecular weight, Mu; by the
`formula:
`
`and of the Test solution at 20°. Calculate the viscosity numbers of
`each of the Test solutions by the formula:
`
`L.W,M,,
`in which W, is the weight proportion of oligosaccharide i; and M, is
`the molecular weight of oligosaccharide i. Use the following
`molecular weight values for calculation:
`
`Glucose
`180
`Isomaltose
`342
`Isomaltotriose
`504
`Isomaltotetraose
`666
`Isomaltopentaose
`828
`Isomaltohexaose
`990
`Isomaltoheptaose
`1152
`Isomaltooctaose
`1314
`Isomaltononaose
`1476
`Isomaltodecaose
`1638
`Isomaltoundecaose
`1800
`Isomaltododecaose
`1962
`Isomaltotridecaose
`2124
`Isomaltotetradecaose
`2286
`Jsomaltopentadecaose
`2448
`Isomaltohexadecaose
`2610
`Isomaltoheptadecaose
`2772
`Isomaltooctadecaose
`2934
`Isomaltononadecaose
`3096
`Calculate the amounts of the fractions with fewer than 3 and with
`more than 9 glucose units for the Reference solution and the Test
`solution: the Mw and amounts of the fractions obtained for the
`Reference solution are within the values stated in the data sheet that
`accompanies USP Dextran l RS. The Mw of Dextran l is between 850
`and 1150. The fraction with fewer than 3 units of glucose is less than
`15%, and the fraction with more than 9 units of glucose is less than
`20%.
`
`Dextran 40
`
`Dextrans.
`Dextrans
`
`[9004-54-0].
`
`» Dextran 40 is derived by controlled hydrolysis and
`fractionation of polysaccharides elaborated by the
`fermentative action of certain strains of Leuconostoc
`mesenteroides (NRRL, B.512 F; NCTC, 10817) on a
`sucrose substrate. It is a glucose polymer in which the
`linkages between glucose units are almost entirely of the
`a-1 : 6 type. Its weight average molecular weight is in the
`35,000 to 45,000 range.
`
`Packaging and storage--Preserve in well-closed containers. Store at
`25°, excursions permitted between 15° and 30°.
`Labeling-Where it is intended for use in preparing injectable
`dosage forms, the label states that it is sterile or must be subjected to
`further processing during the preparation of injectable dosage forms.
`USP Reference standards ( ll )-USP Dextran 40 RS. USP Dextran
`4 Calibration RS. USP Dextran I 0 Calibration RS. USP Dextran 40
`Calibration RS. USP Dextran 70 Calibration RS. USP Dextran 250
`Calibration RS. USP Dextran V, Marker RS. USP Dextran 40 System
`Suitability RS. USP Endotoxin RS.
`Color of solution-The absorbance of a solution in water (1 in 10),
`measured in a 4-cm cell determined at 375 nm against a water blank,
`is not greater than 0.20.
`Identification-
`A:
`Infrared Absorption (197K).
`B: Prepare four Test solutions of Dextran 40 in water, in such a
`manner that the concentrations are accurately known and approxi(cid:173)
`mately evenly distributed in the range of 2% to 0.5%. Using a
`capillary tube viscosimeter having dimensions such that the flow time
`of water is not less than 100 seconds, measure the flow times of water
`
`{ln[(RD)(t I t0)]} I C,
`in which RD is the ratio of the density of the individual Test solution to
`that of water; t and t0 are the flow times for the Test solution and water,
`respectively; and Cis the concentration, in g per mL, of Dextran 40 in
`the Test solution. Plot the viscosity numbers of each of the Test
`solutions against their respective concentrations, and draw the
`straight line of best fit through the points and extrapolate to zero
`concentration: the value ofthe intercept is between 18 and 23 mL per
`g.
`Specific rotation (781S): between +195° and +203°.
`Test solution: 20 mg per mL, heated, if necessary, on a water bath
`to dissolve.
`Bacterial endotoxins (85) (where it is labeled as intended for use in
`the preparation of injectables)- When tested in Sodium Chloride
`Injection (1 in 10), it contains not more than 1.0 USP Endotoxin Unit
`per mL.
`Safety-Inject intravenously 1.0 mL of a sterile 1 in 10 solution of
`10% Dextran 40 in saline TS into each of five mice weighing 18 to
`20 g. The injection period is not less than l 0 seconds and not greater
`than 15 seconds. If there are no deaths within 72 hours, it meets the
`requirements of the test. If I or more animals die, continue the test
`using 10 mice weighing 20 ± 0.5 g. If all animals survive for 72
`hours, the requirements of the test are met.
`pH (791): between 4.5 and 7.0, in a solution (1 in 10).
`Loss on drying (731)-Dry it at 105° for 5 hours: it loses not more
`than 7.0% of its weight.
`Sulfate (221 )-A 1.5-g portion shows no more sulfate than
`corresponds to 0.45 mL of 0.020 N sulfuric acid (0.03%).
`Heavy metals, Method II (231): 5 Jlg per g.
`Limit of nitrogenous impurities (where it is labeled as intended for
`use in the preparation of injectables)-
`Sulfate solution- To 1000 mL of sulfuric acid add 5 g of anhydrous
`cupric sulfate and 500 g of potassium sulfate. Dissolve by heating,
`and store at 60°. [NOTE- If storage at 60° is not possible, prepare a
`smaller quantity of Sulfate solution on the day of use, adjusting the
`proportions accordingly.]
`Indicator-Dilute a mixture of 20 mL of a 0.1% solution of
`bromocresol green in alcohol and 4 mL of methyl red TS with water
`to 100 mL.
`Procedure- Transfer 0.2 g, accurately weighed, to a micro(cid:173)
`Kjeldahl flask. Add 4 mL of Sulfate solution. Heat until the solution
`exhibits a clear green color and the sides of the flask are free from
`carbonaceous material. Cool, and transfer the solution to a steam
`distillation unit. Rinse the Kjeldahl flask three times with 5 mL of
`water, adding the washings to the solution. Add 15 mL of 45%
`sodium hydroxide solution, immediately close the distillation
`apparatus, and commence steam distillation without delay. Receive
`the distillate in 1 mL of Indicator in a 1 00-mL flask, keeping the end
`of the condensing tube below the liquid surface for 5 minutes and
`above the liquid surface for I minute. Upon completion of the
`distillation, remove the receiving flask, and rinse the end of the
`condensing tube with a small quantity of water, adding the rinse to the
`distillate. Titrate the distillate with 0.010 N hydrochloric acid until the
`color changes from blue to reddish violet. Perform a blank
`determination, and make any necessary correction. The corrected
`volume ofO.OlON hydrochloric acid titrated does not exceed 0.14
`mL (0.01 %, as N).
`Limit of alcohol and related impurities-
`Test solution-Dissolve without heating 5.0 gin 100 mL of water,
`and distill the solution, collecting the first 45 mL of the distillate.
`Dilute the distillate with water to 50.0 mL, and mix.
`Standard solution- To 25.0 mL of the Test solution add 0.5 mL of
`a 2.5% (wlv) solution of n-propyl alcohol.
`Chromatographic system-The gas chromatograph is equipped
`with a flame-ionization detector and contains a 2-mm x 1.8-m
`column packed with support S3. The column temperature is
`maintained at about 160°, the injection port temperature is maintained
`at about 240°, and the detector is maintained at about 210°. The
`carrier gas is nitrogen, flowing at a rate of about 25 mL per minute.
`[NOTE- Injector seals may deteriorate after multiple injections of the
`Standard and Test solutions. Inspect the seals before making a series
`of injections.]
`
`
`
`
`IPR2019-01142
`Pharamacosmos v. American Regent
`Petitioner Ex. 1007 - Page 3
`
`