The United States
`Pharmacopeia
`
`TWENTY-FIRST REVISION
`
`Officialfrom January 1, 1985
`
`The National
`Formulary
`
`SIXTEENTH EDITION
`
`Official from January 1, 1985
`
`· ~; ..
`,~ -·• .
`' .
`• ..
`.
`j I
`
`.
`
`. '
`
`• '
`
`.
`
`United States Pharmacopeial Convention, Inc. lU$'®
`
`1260 1 Twinbrook Parkway, Rockville, Md. 20852 ~H =
`
`Eton Ex. 1099
`1 of 9
`
`

`

`NOTICE
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`formerly shown on a coupon in this space,
`is stamped on the spine of the volume.
`
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`I) WAR
`
`I G
`
`Concerning U. S. Patefll or Trademark Rights
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`
`Concerning Use of USP or NF Tex,
`Allcntion is called to the fact that USP and F text is fully copyrighted. Authors and others
`wishing to use portions of the text should request permission 10 do so from the Secretary of the
`SPC Board of Trustees.
`
`© I 984 The United States Pharmacopeia l onvention. Inc.
`1260 1 Twinbrook Parkway, Rockville. Md . 20852
`
`A ll righl.< reserved
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`Eton Ex. 1099
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`

`

`USPXXI
`
`The United States
`Pharn1acopeia
`
`TWENTY-FIRST REVISION
`
`By authority of the United States Pharmacopeial Convention, Inc.,
`meeting at Washington, D. C., April 17- 19, 1980. Prepared by the
`Commillee of Revision and published by the Board of Trustees
`
`Official from January 1, 1985
`
`Un ited States Pharmacopeia l Convent ion, Inc.
`1260 I Twin brook Parkway, Rockville, Md. 20852
`
`Eton Ex. 1099
`3 of 9
`
`

`

`© 1984 T he United States Pharmacopeial Convention, Inc.
`12601 Twinbrook Parkway, Rockville, Md. 20852
`All rights reserved
`
`Eton Ex. 1099
`4 of 9
`
`

`

`This material may be protected by Copyright law (Title 17 U.S. Code)
`
`USP XX/
`
`container, observing precautions against contact with water or moist
`atmosphere. Adjust the concentration of the reagent so_ that the
`titration volume approaches but does not exceed the capacity of the
`container. Titrate to an a mber color that persists for 15 seconds
`after mixing.
`ot more than 1.0% of water is found .
`Other requirement -
`It meets the requirements under Sterility
`Tests ( 71 ) and U111formity of Dosage Units (905).
`Assay-
`Mobile phase - Add 1.03 g of sodium 1-heptancsulfonate to a
`mixtureof900 ml of water and I0mLofmcthanol. Mix, then add
`sufficient glacial acetic acid and ammonium hydroxide, if necessary,
`to adjust the solution to a pH of 4.0. Add SO ml of acetonitrile,
`then add water to make I 000 mL, and mix. Slight variation of the
`amount of acetonitrile may be required to improve resolution or
`adjust retention lime. Dega, the solution.
`Standard preparation- Dissolve an accurately weighed quantity
`of US P Acetylcholinc Chloride RS in Mobile phase, and dilute
`quantitatively and stepwise with Mobile phase to obtain a solution
`having a known concentration about equal to that of the acetyl(cid:173)
`choline chloride in the Assay preparation.
`Assay preparation - Transfer the contents of I container of
`Acetylcholinc Ch loride for Ophthalmic Solution to a JO.-ml volu(cid:173)
`metric nask with the aid of Mobile phase , add Mobile phase to
`volume, and mix .
`hromatographi<' sys1e111 - Usc a liquid chromatograph fitted
`with a 30-cm X 3.9-mm stainless steel column packed with packing
`l I and a refractive index detector. The now rate is about 2 ml
`pe( minute. Chromatograph replicate 50-µl injections of the
`Standard preparation, and record the peak respon e: the relative
`standard deviation is not more than 3.5%. Chromatograph a o(cid:173)
`lution containing abou t 0.2% each of acetylcholine chloride and
`choline chloride: the resolution is not less than 2.0.
`Procedure - Separately inject equal volumes (about 50 µl) of
`the Standard preparatio11 and the Assay preparation into the
`chromatograph, record the chromatograms, and measure the re(cid:173)
`sponses. Calculate the qu:in1i1y, in mg. of C1H 16CINO2 in the
`container taken by the formula I 0C(ru/rs ), in which C is the con(cid:173)
`centration. in mg per mL. of US P Acetylcholinc Chloride RS in the
`Sta11dard preparatio11 , and ru and rs are the peak responses ob(cid:173)
`tained from the Assay preparatio11 and the
`tandard preparation
`respectively.
`
`Acetylcysteine
`
`fCOCH3
`HSCH,· ·· •1 ··· · COOH
`
`H
`
`CsH9 03S
`163.19
`L-Cysteine. N-acetyl- .
`(616-9/-1 j .
`N-Acetyl-L-cysteinc
`» Acetylcysteine contains not less than 98 .0 percent
`and not more than I 02.0 percent of CsH9NO3S, cal(cid:173)
`cu lated on the dried basis.
`Packaging and storage- Preserve in tight containers.
`Reference standard-USP Acetylcysteine Refere11ce Standard(cid:173)
`Dry at a pressure of a bout 50 mm of mercury at 70° for 4 hours
`before using.
`Identification-The infrared absorption spectrum of a mineral oil
`dispersion of it, previously dried, exhibits maxima only at the same
`wavelengths as that of a similar preparation of USP Acctylcysteine
`RS.
`Melting range, Class I (741 ): between 104° and 110°.
`Specific rotation ( 781 )-1 n a 25-ml volumetric nask mix 1.25 g
`with I ml of disodium ethylene-diaminetetraacetate soluuon ( I
`in I 00), add 7 .S ml of sodium hydroxide solution ( I in 25), and mix
`to dissolve. Dilute to volume with pH 7.0 buffer (prepared by
`mixing 29.5 mL of I N sodium hydroxide, 50 ml of I M monobasic
`potassium phosphate, and sufficient water to make 100 ml and,
`using a pH meter, adjusting to a pH of 7.0 ± 0.1 by adding, as
`necessary, more of either solution):
`the specific rotation, calculated
`on the dried basis, compared with a blank prepared with the same
`amounts of the same reagents, is between +21 ° and +27° .
`pH (791 ): between 2.0 and 2.8, in a solution (I in 100).
`
`Official Monographs / Acetylcysteine
`
`19
`
`Loss on drying (731 )-Dry it at a pressure of about 50 mm of
`mercury at 70° for 4 hours:
`it loses not more than 1.0% of its
`weight.
`Residue on ignition (281 ) - Transfer to a tared fused silica dish
`about 2 g, weigh accurately, heat on a hot plate until thoroughly
`charred, cool, add I mL of sulfuric acid, a nd heat gently until
`01
`fuming ceases. Ignite al 600° until the carbon is consumed.
`more than 0.5% is found.
`Hea~y metals, Method II (231) -
`ln a dropwisc manner [Cau(cid:173)
`tion-Exercise care, since explosion may occur), wet the lest
`specimen with 2 ml of nitric acid, and proceed as directed for the
`Test Preparation: the limit is 0.00 I%.
`Assay-
`Mobile phase- Dissolve 6.8 g of monobasic potassium phosphate
`in I 000 ml of water, filter through a membrane filter (0.45-µ po(cid:173)
`rosity), and degas.
`Internal standard solution-Dissolve about I g of di-phenyl(cid:173)
`alanine in 200 ml of freshly prepared sodium bisulfite solution ( I
`in 2000).
`Standard preparation- Dissolve an accurately weighed quantily
`of USP Acetylcysteine RS in sodium bisulfite solution ( I in 2000)
`to obtain a solution having a known concentration of about 10 mg
`per mL. Pipet I 0.0 ml of this solution and 10.0 ml of Internal
`standard solution into a 200-ml volumetric nask, dilute with so(cid:173)
`dium bisulfite solution ( I in 2000) to volume. and mix to obtain a
`Standard preparation having a known concentration of about 0.5
`mg per mL.
`Assay preparation- Transfer about IO0O mg of Acetylcysteinc,
`accurately weighed, to a 100-ml volumetric nask. Dissolve in
`sodium bisulfite solution ( I in 2000), dilute with the same solvent
`to volume, and mix. Pipct 10.0 ml of this olution and 10.0 ml
`of /ntema/ standard ,wlution into a 200-mL volumetric nask, dilute
`with sodium bisulfite solution ( I in 2000) to volume, and mix.
`Chromatographic system (see Chromatography (621) )- The
`liquid chromatograph is equipped with a 214-nm detector and a
`3.9-mm X 30-cm column that contains packing l I. The now rate
`is about 1.5 ml per minute. Chromatograph the Standard prep(cid:173)
`aration, and record the peak responses as directed under Procedure:
`the relative standard deviation for replicate injections is not more
`than 2.0% and the resolution, R. factor between acctylcystcinc and
`di-phenylalanine is not less than 6.
`Procedrire- Scparalcly inject equal volumes (about S µL) of the
`Standard preparation and the Assay preparation into the chro(cid:173)
`matograph, record the chromatograms, and measure the responses
`for the major peaks. The relative retention times are about 0.4 for
`acctylcysteinc and J .0 for di-phenylalanine. Calculate the quantity,
`in mg, of C 5H9NO3S in the Acetylcysteine taken by the formula
`2000C(Ru/Rs), in which C is the concentration, in mg per mL. of
`USP Acetylcysteine RS in the Standard preparation , and Ru and
`Rs are the ratios of the peak response of acetylcysteine to that of
`di-phenylalanine obtained from the Assay preparation and the
`Standard preparation, respectively.
`
`Acetylcysteine Solution
`
`» Acetylcystcine Solution is a sterile solution of Ace(cid:173)
`tylcystcine in water, prepared with the ai d of Sodium
`It contains not less than 90.0 percent and
`Hydroxide.
`not more than 110.0 percent of the labeled amount of
`CsH9 O3S.
`Packaging and storage-Preserve in single-dose or multiple-dose
`containers, preferably of Type l glass, tightly closed with a glass or
`polyethylene closure.
`Reference standard-USP Acetylcysteine Reference Standard(cid:173)
`Dry at a pressure of about 50 mm of mercury at 70° for 4 hours
`before using.
`Identification-Place 2 ml in a 10-mL beaker, and adjust to a pH
`of about 3 (pH indicator paper) using 3 N hydrochloric acid. Add
`500 mg to J g of finely powdered sodium chloride, in two portions
`of about 200 mg each initially, and then in smaller portions (about
`25 mg), stirring after each addition, until a precipitate is formed .
`Allow to stand at room temperature for 15 minutes, and collect the
`residue by suction filtration. The acetylcysteine so obtained,
`after being dried as directed in the lest for Loss on drying under
`
`Eton Ex. 1099
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`
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`20
`
`Acetylcysteine / Official Monographs
`
`Acetylcysteine, responds to the Identification res/ under Acetyl(cid:173)
`cysteine.
`Sterility-
`It meets the requirements under Sterility Te sis (71 ) .
`pH ( 791 ) : bet ween 6.0 and 7 .5.
`Assay-
`Mobile phase, /111ernal standard solution, and Standard prep(cid:173)
`aration- Prepare as directed in the Assay under Ace1ylcys(cid:173)
`teine.
`Assay preparation- Pipe! a volume of Acetylcysteinc Solution,
`equivalent lo about I 000 mg of acetylcysteine, into a I 00-mL vol(cid:173)
`umetric nask, dilute with sodium bisulfite solution ( I in 2000) to
`volume, and mix. Pi pet I 0.0 ml of this solution and I 0.0 mL of
`flllernal s1andard solution into a 200-ml volumetric n ask, dilute
`with sodium bisulfite solution ( I in 2000) to volume, and mix.
`Chromatographic system - Proceed as directed for Chromaro(cid:173)
`graphic system m the Assay under Acetylcysteine.
`Procedure- Proceed as directed for Procedure in the Assay
`under Acetylcysteine. Calculate the quantity, in mg, of C5H9NO3S
`in each ml of the Solution taken by the formula 2000(C/V)(Ru/
`Rs), in which C is the concentration, in mg per ml, ·of USP Ac(cid:173)
`etylcysteine RS in the Standard preparation, Vis the volume, in
`mL, of Solution taken, and Ru and Rs are the ratios of the peak
`response of acetylcysteine to that of di-phenylalanine obtained from
`the Assay preparation and the Standard preparation. respec(cid:173)
`tively.
`
`Acetylcysteine and Isoproterenol
`Hydrochloride Inhalation Solution
`» Acetylcysteine and lsoproterenol Hydrochloride
`Inhalation Solution is a sterile solution of Acetylcys(cid:173)
`teine and Isoproterenol Hydrochloride in water. It
`contains not less than 90.0 percent and not more than
`110.0 percent of the labeled amount of acetylcysteine
`(CsH9NO3S), and not less than 90.0 percent and not
`more than 115.0 percent of the labeled amount of iso(cid:173)
`proterenol hydrochloride (Ct tH 17NO3 • HCI).
`Packaging and storage- Preserve in single-dose or in multiple-dose
`containers, preferably of Type I glass, tightly closed with a glass or
`polyethylene closure.
`Reference standards- USP Acetylcysteine Reference Stan(cid:173)
`dard- Dry at a pressure of about 50 mm of mercury at 70° for 4
`hours before using. USP lsoproterenol Hydrochloride Reference
`Standard-Dry in vacuum over phosphorus pentoxide for 4 hours
`before using.
`Identification-
`A: Place 2 mL in a I 0-ml beaker, and adjust with 3 N hydro(cid:173)
`chloric acid to a pH of about 3 (pH indicator paper). Add 500 mg
`lo I g of finely powdered sodium chloride, in two portions of about
`200 mg each initially, and then in smaller portions (about 25 mg).
`stirring after each addition, until a precipitate is formed. Allow
`to stand at room temperature for 15 minutes, and collect the residue
`by suction filtration : the acetylcysteine so obtained, after being
`dried as directed in the test for Loss 011 dryi11g under Acetylcysteine.
`responds to the Identification test under Acetylcysteine.
`B: Ferro-citrate solution and Buffer solution-Prepare as
`directed under Epinephrine Assay ( 391).
`Procedure- Place a volume of Inhalation Solution, equivalent
`to about 0.26 mg of isoproterenol hydrochloride, in a test tube with
`3 m L of 0.1 M mercuric chloride, and mix. Add I 00 µL of
`Ferro-citrate solution and 1.0 mL of Buffer solution, and mix:
`the
`presence of isoproterenol hydrochloride is confirmed by the devel(cid:173)
`opment of a purple color.
`Sterility-It meets the requirements under Sterility Tests (7 I).
`pH (791): between6.0and7.0.
`Assay for acetylcysteine-
`Mobi/e phase, Jnternal standard solution, and Standard prep(cid:173)
`aration-Prepare as directed in the Assay under Acetylcysteine.
`Assay preparation- Pipet a volume of Acetylcysteine and Jso(cid:173)
`proterenol Hydrochloride Inhalation Solution, equivalent to about
`I 000 mg of acetylcystcine, into a I 00-mL volumetric nask, dilute
`with sodium bisulfite solution ( I in 2000) to volume, and mix. Pi pet
`
`USP XX/
`
`IO mL of this solution and 10 mL of Internal standard solutio11 into
`a 200-mL volumetric nask, dilute with sodium bisulfite solution ( I
`in 2000) to volume, and mix.
`Chromatographic system- Proceed as directed for Chromaro(cid:173)
`graphic system in the Assay under Acetylcysteine.
`Procedure- Proceed as directed for Procedure in the Assay
`under Acetylcysteine. Calculate the quantity, in mg, of CsH9NO3S
`in each mL of the Inhalation Solution taken by the formula
`2000(C/V)(Ru/Rs), in which C is the concentration, in mg per ml,
`of USP Acetylcysteine RS in the Srandard preparation, Vis the
`volume, in ml, of Inhalation Solution taken, and Ru and Rs are
`the ratios of the peak response of acetylcysteine to that of di-phe(cid:173)
`nylalanine obtained from the Assay preparation and the Standard
`preparation, respectively.
`Assay for isoproterenol hydrochoride-
`Mobi/e phase-Dissolve 13.6 g of monobasic potassium phos(cid:173)
`phate in 1000 ml of water, and filter through a membrane filter
`(0.45-µm porosity). Add 20.0 ml of methanol, mix, and degas.
`Internal standard solution- Place about 150 mg of acetami(cid:173)
`nophen in a 500-ml volumetric nask, add 5 mL of glacial acetic
`acid, dilute with water to volume, and mix.
`Standard preparation-Transfer about 30 mg of USP lsopro(cid:173)
`terenol Hydrochloride RS, accurately weighed, to a 200-ml volu(cid:173)
`metric nask. Add 200 mg of sodium bisulfite, dilute with water
`to volume, and mix. Pipct 10 mL of this solution and 10 mL of
`Internal sra11dard solutio11 into a 25-ml volumetric flask, add dilute
`glacial acetic acid ( I in I 00) to volume, and mix to obtain a Stan(cid:173)
`dard preparation having a known isoprotcrenol hydrochloride
`concentration of about 0.06 mg per ml.
`Assay preparation-Transfer an accurately measured volume
`of Inhalation Solution, equivalent to about 1.5 mg of isoprotcrenol
`hydrochloride, and 10 mL of lllfernal standard solution to a 25-mL
`volumetric flask, add dilute glacial acetic acid ( I in I 00) to volume,
`and mix.
`Chromatographic system (see Chromatography ( 621) )- The
`liquid chromatograph is equipped with a 280-nm detector and a
`3.9-mm X 40-cm column that contains packing LI . The flow rate
`is about 2 mL per minute. Chromatograph the Sta11dard prepa(cid:173)
`ration, and record the peak responses as directed under Procedure:
`the relative standard deviation for replicate injections is not more
`than 2.0% and the resolution, R . between isoproterenol hydro(cid:173)
`chloride and acetaminophen is not less than 6.
`Procedure-Separately inject equal volumes (about 25 µl) of
`the Standard preparation and the Assay preparario11 into the
`chromatograph, record the chromatograms. and measure the re(cid:173)
`sponses for the major peaks. The relative retention times are about
`0.5 for isoproterenol hydrochloride and 1.0 for acetaminophen.
`Calculate the quantity, in mg, of C1 1 H 17NO3. HCI in each mL of
`the Inhalation Solution taken by the formula (25C/V)(Ru/Rs), in
`which C is the concentration. in mg per ml, of USP lsoproterenol
`Hydrochloride RS in the Sta11dard preparation, Vis the volume,
`in ml, of Inhalation Solution taken. and Ru and Rs are the ratios
`of the peak responses of isoproterenol hydrochloride to those of
`acetaminophen obtained from the Assay preparation and the
`Standard preparation, respectively.
`
`Acids-see complete list in index
`
`Acrisorcin
`
`© lOO •~OH
`CH,ICH,l.CH,
`
`388.51
`C12H 1sO2.CuH10N2
`1,3-Benzenediol, 4-hexyl-, com pd. with 9-acridinamine (I: I).
`4-Hexylresorcinol compound with 9-aminoacridine (I: I)
`.
`[7 527-9 !-5].
`» Acrisorcin contains not less than 97.0 percent and
`not more than 101.0 percent ofC12H1sO2.C13H10N2,
`calculated on the dried basis.
`Packaging and storage- Preserve in well-closed containers.
`
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`268
`
`Cysteine / Official Monographs
`
`Assay-
`Mixed so/ve111- Mix 10 mL of methanol, 2 drops of hydrochloric
`acid, and suflicient chloroform (saturated with water) lo make I 00
`ml.
`S1andard preparation- Dissolve a suitable quantity of USP
`Cyprohcpladinc Hydrochloride RS, accurately weighed, in the
`Mixed solve,,/, and prepare, by quantitative and stepwise dilution,
`a solution in Mixed solvem containing about 20 µg per m L.
`Chromatographic column- Proceed as directed under Column
`Partition Chromatography (621 ), packing;i chromatographic tube
`with two segments of packing material. The lower segment is a
`mixture of 2 g of Solid Support and I mL of sulfamic acid solution
`( I in 20), and the upper segment is a mixture prepared as directed
`under Assay preparation.
`Assay preparation-Weigh and finely powder not less. than 20
`Cyproheptadine Hydrochloride Tablets. Transfer an accurately
`weighed portion of the powder, equivalent lo about 4 mg of cypro(cid:173)
`heptadine hydrochloride, to a 100-mL beaker, add I ml of spec(cid:173)
`trophotometric grade dimethyl sulfoxide, and mix to wet the powder
`evenly. Add I ml of sulfamic acid solution ( I in 20), mix, add 3
`g of Solid Support. mix as directed under Chromatographic col(cid:173)
`umn, and transfer to the column.
`Procedure-Wash the column with 100 mL of ether (saturated
`with water), and discard the washing. Place under the column, as
`a receiver, a 200-ml volumetric nask containing 20 ml of methanol
`and 4 drops of hydrochloric acid. Pass through the column 150 mL
`of chloroform (saturated with water). Mix the eluate, allow to cool
`to room temperature, dilute with chloroform (saturated with water)
`to volume, and mix. Concomitantly determine the absorbances of
`this solution and of the Standard preparation in I-cm cells at the
`wavelength of maximum absorbancc at about 286 nm, with a suit(cid:173)
`able spectrophotometer, using the Mixed solvent as the blank.
`Calculate the quantity, in mg, of C21 H2, N. HCI in the portion of
`Tablets taken by the formula 0.2C(Au/As ), in which C is the con(cid:173)
`centration, in µg per ml, of USP Cyproheptadine Hydrochloride
`RS in the Standard preparation, and Au and As arc the absorb(cid:173)
`ances of the solution from the Assay preparation and the Standard
`preparation, respectively.
`
`Cysteine Hydrochloride
`
`~
`HSCH,- ~- COOH •
`
`'1CI • H,O
`
`NH,
`
`(7048-04-6).
`
`175.63
`C1H1NO2S. HCI. 1-120
`L-Cysteine hydrochloride monohydrate.
`L-Cysteine hydrochloride monohydratc
`[52-89-/ J.
`Anhydrous
`157.61
`» Cysteine Hydroch loride contains not less than 98.5
`percent and not more than 101.5 percent of C3H7·
`N02S. HCI, as L-cysteine hydrochloride, calculated on
`the dried basis.
`Packaging and storage- Preserve in well-closed containers.
`Reference standard-USP L·C)'steine Hydrochloride Reference
`Standard- Dry at a pressure of 5 mm of mercury for 24 hours be(cid:173)
`fore using.
`Identification- The infrared absorption spectrum of a potassium
`bromide dispersion of it. previously dried, exhibits maxima only at
`the same wavelengths as that of a similar preparation of USP L·
`Cysteinc Hydrochloride RS.
`S pecific rotation (781 ): between +5.7° and +6.8°, calculated on
`the dried basis, determined in a solution in 6 N hydrochloric acid
`containing 800 mg in each 10 ml.
`Loss on drying (73 1 }- Dry it at a pressure of 5 mm of mercury for
`24 hours:
`it loses between 8.0% and 12.0% of its weight.
`Residue on ignition ( 28 1 ) : not more than 0.4%.
`Chloride content (22 1 ) - Dissolve about 350 mg, accurately
`weighed, in IO mL of dilute nitric acid (I: I). Add 30.0 m L ofO. l
`N silver nitrate VS. Add 40 ml of saturated potassium perman(cid:173)
`ganate solution. Heat for JO minutes on a steam bath, and cool.
`Add 30 percent hydrogen peroxide dropwise until the solution be(cid:173)
`comes colorless. Add 8 ml of ferric ammonium sulfate TS and I
`
`USP XX!
`
`ml of nitrobcnzene. Titrate with 0.1 N ammonium thiocyanate
`VS. Each ml of 0.1 N silver nitrate is equivalent to 3.545 mg of
`chloride: between 19.8% and 20.8% is found.
`Sulfate (221 )- A solution containing 0.33 g shows no more sulfate
`than corresponds to 0. 10 m L of 0.020 N sulfuric acid (0.03%).
`Arsenic (211 ): 1.5 ppm.
`Iron ( 241 ) : 0.003%.
`Hea~y metals, Method I (231 ): 0.00 15%.
`Assay- Weigh accurately about 250 mg of Cystcine Hydrochloride
`into an iodine nask. Add 20 ml of water and 4 g of potassium io(cid:173)
`dide, and mix to dissolve. Cool the solution in an ice bath, and add
`5 mL of 3 N hydrochloric acid and 25.0 mL of 0.1 N iodine VS.
`Insert the stopper, and allow to stand in the dark for 20 minutes.
`Titrate the excess iodine with 0.1 N sodium thiosulfate VS, adding
`3 mL of starch TS llS the end-point is approached. Perform a blank
`determination, and make any necessary correction. Each ml of
`0.1 N
`iodine is equivalent to 15.76 mg of CJH1NO2S.(cid:173)
`HCI.
`
`Cysteine Hydrochloride Injection
`
`» Cystcinc Hydrochloride Injection is a sterile solution
`of Cysteine Hydrochloride in Waler for Injection. It
`contains not less than 85.0 percent and not more than
`115.0 percent of the labeled amount of C3H7N02S .(cid:173)
`HCI.H 20.
`Packaging and storage-Preserve in single-dose or in multiple-dose
`containers, preferably of Type I glass.
`Reference standard-USP L-Cysteine llydrochloride Reference
`Sumdard- Dry at a pressure of 5 mm of mercury for 24 hours be(cid:173)
`fore using.
`Identification-
`A: To2mLoflnjcctionadd3mLofwater,andmix. Add 10
`mL of cupric nitrate TS: a bluish gray precipitate is formed.
`B: To 2 ml of Injection add 3 ml of water, and mix. Add 2
`m L of 3 N sodium hydroxide and 2 drops of sodium nitroferricya(cid:173)
`nide solution ( I in 20): a red-purple color is produced. and it rapidly
`changes to yellow.
`C:
`It responds Lo the tests for Chloride ( I 9 I).
`lt meets the requirements of the Pyrogen Test ( 151).
`Pyrogen-
`pll (791 ): between 1.0 and 2.5.
`Heafy metals, Method fl (231 ): 2 ppm.
`It meets the requirements under Injections
`Other requirements-
`( I ) •
`Assay-
`Standard solu1io11- Dissolve an accurately weighed quantity
`of USP L-Cysteine Hydrochloride RS in nitrogen-saturated water
`to obtain a solution having a known concentration of about I mg
`per ml..
`Standard preparation- Transfer 20.0 ml of Standard solution
`to a 200-ml volumetric nask. dilute with nitrogen-saturated 1.0
`N sodium hydroxide to volume, and mix.
`Assay preparation- Dilute an accurately measured volume of
`Cysteine Hydrochloride Injection, equivalent to about 250 mg of
`cysteine hydrochloride, quantitatively and stepwise with nitro(cid:173)
`gen-saturated 1.0 N sodium hydroxide. to obtain a solution having
`a concentration of about 0.1 mg per m L.
`Procedure- Transfer a suitable amount of Standard preparation
`to a polarographic cell. With mercury dropping from the electrode,
`lower the dropping mercury electrode of a polarograph so that the
`end is submerged in the liquid. Bubble oxygen-free. water-satu(cid:173)
`rated nitrogen through the liquid for 15 minutes. Record the
`polarogram from -0.2 volt to -1. 10 volts. using a saturated calomel
`electrode as the reference electrode. In a similar manner. record
`the polarograms obtained using portions of the Assay preparation
`and of the nitrogen-saturated 1.0 N sodium hydroxide. Determine
`the height of the diffusion current wave at -0.4 volt. Calculate the
`quantity. in mg, of C3~hNO~S . HCI. H2O in each ml of the In(cid:173)
`jection taken by the formula 2500((./V)[(i,1)u/(ic1)s]. in which C
`is the conccntrat1on. in mg per ml, of USP L-Cystcine Hydro(cid:173)
`chloride RS in the Standard preparation, Vis the volume, in ml.
`of Injection taken. and (i,,)u and (i,1)s arc the obscrvcd diffusion
`
`Eton Ex. 1099
`7 of 9
`
`

`

`USP XX/
`
`currents, corrected for the diffusion current of the 0.1 N sodium
`hydroxide. of the Assay preparation a nd the S tandard preparation.
`respectively.
`
`Cytarabine
`
`243.22
`C9H 13N3(cid:144)
`5
`2( I H)-Pyrimidi none, 4-amino-1-.B-o-arabinofuranosyl-.
`[ 147-94-4).
`1-/3-D-Arabinofuranosylcytosine
`» Cytarabine contains not less tha n 95.0 percent a nd
`not more than 105.0 percent of C9H 13N30s, calculated
`o n the dried basis.
`Packaging and storage- Preserve in well-closed, light-resistant
`containers.
`Reference standard- USP Cytarabine Reference Standard(cid:173)
`[ Caution- Avoid contact). Dry at a pressure not exceeding 5 mm
`of mercury al 60° for 3 hours before using.
`Jdentification-
`A: The infrared absorption spectrum of a mineral oil dispersion
`of it. previously d ried at a pressure of not mo re than 5 mm of mer(cid:173)
`cury at 60° for 3 hours, exhibits maxima only at the same wave(cid:173)
`lengths as that of a similar preparation of USP Cytarabine RS .
`B: The ultraviolet a bsorption spectrum of a I in I 00.000 solu(cid:173)
`tion of it in 0. 12 N hyd rochloric acid exhibits maxima and minima
`at the same wavelengths as that of a similar solution of USP Cy(cid:173)
`tarabinc RS, concomitantly measured. and the respective absorp(cid:173)
`tivities. calculated on the dried basis. at the wavelength of maximum
`absorbance at about 279 nm do not differ by more than 3.0%.
`Specific rotation (78 1): between +154° and +160°.calculated
`on the dried basis. determined in a solution in water containing l 00
`mg in each 10 ml.
`Losson drying (731 )- Dry it in vacuum at a pressure not exceeding
`5 mm of mercury at 60° for 3 hours:
`it loses not more than 1.0%
`of its weight.
`Nitrogen content-Weigh accurately about 200 mg, transfer to a
`Kjeldahl nask, and determine the nitrogen content as directed under
`Nitrogen Determination (46 1 ), using selenium instead of cupric
`sulfate, and using 0.1 N sulfuric acid VS as the titrant. Each mL
`ofO. l N acid is equivalent to 1.40 I mg of N. Not less tha n 16.8%
`and not more than 17.8% of N , calculated on the dried basis, is
`found.
`Residue on ignition (281 ): not more than 0.5%.
`Assay-
`Standard preparation- Dissolve a suitable quantity of US P
`Cytarabine RS , accurately weighed, in water, and dilute quanti(cid:173)
`tatively and stepwise with water 10 obtain a solution having a known
`concentration of abou t 5 mg per mL.
`Assay preparation- Weigh accurately a bout 125 mg of Cytar(cid:173)
`abine, transfer to a 25-mL volumetric nask. dissolve in water, dilute
`wi th water to volume, a nd mix.
`Procedure- Divide the area of a suitable thin-layer chromato(cid:173)
`graphic plate. coated with a 0.5-mm layer of chromatographic silica
`gel mixture into three equal sections. the left and right sections to
`be used for the Assay preparation and the Standard preparation.
`respectively, a nd the center section for the blank. Just prior to use,
`wash the chromatographic plate in a solvent mixture consisting of
`benzene and n-propyl alcohol ( l : I). After allowing the pre-wash
`solvents to evaporate, apply 35 µLeach of the Standard preparation
`and the Assay preparation as streaks 2.5 cm from the botlom of the
`appropriate sections of the plate, drying each solution as it is a pplied
`with a stream of cool air. Using a freshly prepared solvent system
`consisting of methyl ethyl ketone-acetone-water (65:20: I 5). develop
`the chromatogram in a suitable chamber. previously equilibrated
`a nd lined with absorbent paper. until the solvent front has moved
`I 5 cm above the initial streaks. Remove the plate. evaporate the
`
`Official M onographs / Dacarbazine
`
`269
`
`solven t under an air current, and locate the principal band of the
`Standard preparation by viewing under short-wavelength ultravi(cid:173)
`olet light. Mark the a rea of this band, as well as the areas of the
`corresponding bands in the Assay preparation a nd blan k sections
`of the plate. R emove the silica gel from each area separately by
`suitable means, and transfer it to a glass-stoppered, 50-mL conical
`nask. T o each nask add 25.0 ml ofO.0 I N alcoholic sulfuric acid,
`insert the stopper, and shake on a platform shaker for 30 minutes.
`Transfer a portion of each solution to a centrifuge tube. centrifuge
`for 5 minutes, and withd raw the clear, supernatant solution.
`Concomitantly determine the absorbanccs of the solutions from the
`Assay preparation and the Standard prepararion at the wavelength
`of maximum absorbance at about 285 nm in 1-cm cells, with a
`suitable spectrophotometer, against the solution obtained from the
`plate blank. Calculate the q uantity, in mg, of C9H 13N 3O5 in the
`portion of Cytarabine taken by the formula 25C(Au/ As), in which
`C is the concentration, in mg per mL. of USP Cytarabine RS in the
`Srandard preparation, and A u and As are the a bsorbances of the
`solutions from the Assay preparation a nd the S tandard prepara(cid:173)
`tion. respectively.
`
`Sterile Cytarabine
`» Sterile Cytarabine is Cytarabine suitable for pa(cid:173)
`renteral use. It contains not less than 90.0 percent and
`not more than I 10.0 percent of the labeled amount of
`C9H13N305.
`Packaging and storage- Preserve in Containers for Sterile Solids
`as described under Injections ( t).
`Reference standard- USP Cytarabine Reference Srandard(cid:173)
`[ Cautio11- Avoid comact ]. Dry at a pressure not exceeding 5 mm
`of mercury at 60° for 3 hours before using.
`Constituted solution-At the time of use, the constituted solution
`prepared from Sterile Cytarabine meets the requirements for
`Co11s1ituted So/11tio11s under Injections ( I ) .
`Identification- The Assay preparation and the Standard prepa(cid:173)
`ration prepared as directed in the Assay exhibit zones of comparable
`R1 values.
`pH (791 ): between 4.0 and 6.0, in a solutio n containing the
`equivalent of IO mg of cytarabine perm L.
`Water, Method I (92 1): not more than 3.0%.
`Other requirements-
`It meets the requirements for Steriliry Tests
`(7 l ) , Uniformity of Do.rnge Units (905), and Labeling under In(cid:173)
`jections ( I ) •
`Assay- Proceed with Sterile Cytarabine as directed in the Assay
`under Cy1arabi11e.
`
`Dacarbazine
`
`182.18
`C 6 H 10N6O
`I /-/-lmidazolc-4-carboxamidc. 5-(3,3-dimethyl- 1-triazenyl)-.
`5-(3.3- Dimcthyl-1-triazeno)imidazole-4-carboxamidc
`f 4342-03-04 1-
`» Dacarbazinecontains not less tha n 97.0 percent and
`not more than I 02._0 percent of C 6H 1oN60 .
`Caution - Great care should be 1C1ke11 in handling
`Dacarbazine, as it is a potent cytotoxic age/11.
`Packaging and storage-Preserve in