480
`
`Ciprofloxacin / Official Monographs
`
`USP28
`
`USP28
`
`and record the responses as directed under Procedure: the capacity
`factor, le, for the ciprofloxacin peak is between 1.5 and 6, the column
`efficiency is not less than 500 theoretical plates, the tailing factor for
`the analyte peak is not less than 0.9 and not more than 2.0, and the
`relative standard deviation for replicate injections is not more than
`2%.
`Procedure-Separately inject equal volumes (about 20 µL) of the
`Standard preparation and the Assay preparation into the chromat(cid:173)
`ograph, record the chromatograms, and measure the areas for the
`ciprofloxacin peaks. Calculate the quantity, in mg, of ciprofloxacin
`(C 17H11FN30 3) in each mL of the Ophthalmic Solution taken by the
`formula:
`
`(331.34 / 367 .81 )(SOC/ V)(rulrs),
`in which 331.34 and 367 .8 I are the molecular weights of
`ciprofloxacin and anhydrous ciprofloxacin hydrochloride, respec(cid:173)
`tively; C is the concentration, in mg per mL, of USP Ciprofloxacin
`Hydrochloride RS in the Standard preparation, calculated on the
`anhydrous basis; V is the volume, in mL, of Ophthalmic Solution
`taken; and ru and rs are the peak responses obtained from the Assay
`preparation and the Standard preparation, respectively.
`
`Ciprofloxacin Tablets
`
`» Ciprofloxacin Tablets contain Ciprofloxacin Hydro(cid:173)
`chloride equivalent to not less than 90.0 percent and not
`more than 110.0 percent of the labeled amount of
`ciprofloxacin (C 17H1sFN3O3).
`Packaging and storage-Preserve in well-closed containers.
`USP Reference standards ( 11 )-USP Ciprojloxacin Hydrochloride
`RS. USP Ciprofloxacin Ethylenediamine Analog RS.
`Identification-
`A: The retention time of the major peak in the chromatogram of
`the Assay preparation corresponds to that in the chromatogram of the
`Standard preparation, as obtained in the Assay.
`B: Place a number of Tablets, equivalent to about 1500 mg of
`ciprofloxacin, in a suitable flask containing about 750 mL of water,
`and sonicate for about 20 minutes. Dilute with water to I 000 mL, and
`mix. Centrifuge a portion of this suspension, and use the clear
`supernatant obtained as the test solution. Dissolve a quantity of USP
`Ciprofloxacin Hydrochloride RS in water to obtain a Standard
`solution containing 1.5 mg per mL. Proceed as directed for
`Identification test B under Ciprofloxacin Hydrochloride, starting
`with "Separately apply, as I-cm bands, 5 µLeach," except to use 10
`µL each of the test solution and the Standard solution: the specified
`result is obtained.
`Dissolution (711)-
`Medium: 0.01 N hydrochloric acid; 900 mL.
`Apparatus 2: 50 rpm.
`Time: 30 minutes.
`Procedure-Determine the amount of ciprofloxacin hydrochloride
`(C 17H.,FN30 3 • HCI) dissolved by employing UV absorption at the
`wavelength of maximum absorbance at about 276 nm on filtered
`portions of the solution under test, suitably diluted with Dissolution
`Medium, if necessary, in comparison with a Standard solution having
`a known concentration of USP Ciprofloxacin Hydrochloride RS in
`the same Medium.
`Tolerances-An amount of ciprofloxacin hydrochloride
`(C 17H 11FN30 3 • HCI) equivalent to not less than 80% (Q) of the
`labeled amount of ciprofloxacin (C 17H 11FN30 3) is dissolved in 30
`minutes.
`Uniformity of dosage units (905): meet the requirements.
`Assay-
`Diluent-Prepare a filtered and degassed mixture of 0.025 M
`phosphoric acid, previously adjusted (with triethylamine) to a pH of
`2.0 ± 0.1, and acetonitrile (87: 13).
`Mobile phase-Prepare a filtered and degassed mixture of 0.025 M
`phosphoric acid, previously adjusted (with triethylamine) to a pH of
`3.0 ± 0.1, and acetonitrile (87: 13). Make adjustments if necessary
`(see System Suitability under Chromatography {621 )).
`
`~tandard p~eparation-:Quantitat\vely dissolv~ an ~ccurately
`weighed quantity of USP C1profloxacm Hydrochlonde RS m Diluent
`to obtain a solution having a known concentration of about 0.2 mg
`per mL.
`Resolution solution-Dissolve a quantity of USP Ciprofloxacin
`Ethylenediamine Analog RS in the Standard preparation to obtain a
`solution containing about 0.05 mg per mL.
`Assay preparation-Transfer 5 Tablets to a 500-mL volumetric
`flask, add about 400 mL of Diluent, and sonicate for about 20
`minutes. Dilute with Diluent to volume, and mix. Quantitatively
`dilute an accurately measured volume of this solution, previously
`filtered through a 0.45-µm membrane filter, with Diluent to obtain a
`solution containing the equivalent of about 0.20 mg of ciprofloxacin
`per mL.
`Chromatographic system (see Chromatography (621))--The
`liquid chromatograph is equipped with a 278-nm detector and a
`4.6-mm x 25-cm column that contains packing LI and is operated at
`30 ± 1°. The flow rate is about 1.5 mL per minute. Chromatograph
`the Resolution solution, and record the peak responses as directed for
`Procedure: the retention time for ciprofloxacin is between 6.4 and
`I 0.8 minutes; the relative retention times are about 0. 7 for
`ciprofloxacin ethylenediamine analog and 1.0 for ciprofloxacin; and
`the resolution, R. between the ciprofloxacin ethylenediamine analog
`peak and the ciprofloxacin peak is not less than 6. Chromatograph the
`Standard preparation, and record the peak responses as directed for
`Procedure: the column efficiency, determined from the ciprofloxacin
`peak, is not less than 2500 theoretical plates; the tailing factor for the
`ciprofloxacin peak is not more than 2.0; and the relative standard
`deviation for replicate injections is not more than 1.5%.
`Procedure-Separately inject equal volumes (about 10 µL) of the
`Standard preparation and the Assay preparation into the chromat(cid:173)
`ograph, record the chromatograms, and measure the peak areas for the
`major peaks. Calculate the quantity, in mg, of ciprofloxacin
`(C 17H,.FN30 3) in each Tablet taken by the formula:
`(331.34/367.81 )( CLID)(rul rs),
`in which 331.34 and 367 .8 I are the molecular weights of
`ciprofloxacin and anhydrous ciprofloxacin hydrochloride, respec(cid:173)
`tively; C is the concentration, in mg per mL, of USP Ciprofloxacin
`Hydrochloride RS in the Standard preparation, calculated on the
`anhydrous basis; L is the labeled quantity, in mg, of ciprofloxacin in
`each Tablet; D is the concentration, in mg per mL, of ciprofloxacin in
`the Assay preparation, based on the labeled quantity per Tablet and
`the extent of dilution; and ru and rs are the ciprofloxacin peak areas
`obtained from the Assay preparation and the Standard preparation,
`respectively.
`
`Cisplatin
`
`CI,H.N,Pt 300.04
`Platinum, diamminedichloro-, (SP-4-2)-.
`[I 5663-27-1].
`cis-Diamminedichloroplatinum
`
`» Cisplatin contains not less than 98.0 percent and not
`more than 102.0 percent ofC!iH6N2Pt, calculated on the
`anhydrous basis.
`Caution-Cisplatin is potentially cytotoxic. Great care
`should be taken to prevent inhaling particles and
`exposing the skin to it.
`
`Packaging and storage-Preserve in tight containers. Protect fiom
`light.
`USP Reference standards ( 11 )-USP Cisplatin RS. USP Transpla(cid:173)
`tin RS. USP Potassium Trichloroammineplatinate RS.
`
`Jdentification-
`A: The retention tiJ
`the Assay preparatio
`preparation as obtainec
`B:
`Infrared Absorp
`C: Spray reagent(cid:173)
`hydrochloric acid, and
`that all of the solids dis
`90 mL of water. Mix
`precipitate that is forme(cid:173)
`at least I week.
`Procedure-Prepare
`per mL and a Standard ~
`per mL, both in din
`quantities of each solt
`coated with a 0.25-mm
`(see Chromatograph}
`chromatographic charr
`equilibrated for 30 mim
`of acetone and I N nit
`distance of about 8 cm fi
`to air-dry. Complete th
`about I 00° for I minute.
`an oven at about I Q0° l
`solution of potassium io
`spots: the principal Sf
`appearance and RF valu
`Crystallinity (695):
`r
`Water, Method I (921):
`UV purity ratio--{N01
`hydrochloric acid and n
`and dry before use. Do
`acetone or pressurized ,
`light, and use within I h
`mg of ground Cisplatin
`hydrochloric acid to ,
`alternately stir at a big
`seconds until complet,
`frequently to remove pa
`UV absorption spectrur
`0.1 N hydrochloric aci
`absorbance at the maxin
`246 nm is not less than
`Limit of trichloroamm
`Mobile phase-Trani
`volumetric flask, dissol•
`Degas, and filter throu~
`this solution is 5.9 ± 0.
`the Mobile phase, if
`requirements.
`Standard preparatio1
`solve a suitable quantity
`RS, accurately weighed
`saline TS to obtain a sol
`6 µg per mL. Use withi
`Test preparation-[t(cid:173)
`about 50 mg of Cis1
`volumetric flask, and d
`dissolve by stirring by r
`4 hours.
`. Chromatographic s:
`hquid chromatograph ,
`4.6-mm x 25-cm colun
`about 2 mL per minuti
`and record the peak r
`resolution, R, between ·
`platinate peak is not Jes.
`for replicate injections i
`Procedure-Separate
`Standard preparation i
`graph, record the chrom
`due to trichloroammin
`
`Slayback Exhibit 1055, Page 20 of 78
`Slayback v. Eye Therapies - IPR2022-00142
`
`

`

`USP 28
`
`usP28
`
`Official Monographs I Crotamiton
`
`551
`
`ldentification-
`Infrared Absorption ( l 97F}.
`A:
`B: Ultraviolet Absorption ( l 97U)(cid:173)
`Solution: 20 µg per mL.
`Medium : cyclohexane.
`C: To about 10 mL of a saturated solution in water add a few
`drops of potassium permanganate TS: a brown color is produced, and
`a brown precipitate is formed on standing.
`Specific gravity (841 ): between 1.008 and 1.011 at 20°.
`Refractive index (831 ): between 1.540 and 1.543 at 20° .
`Residue on ignition (281 ): not more than 0.1%.
`Bound halogen-Place 4 drops in a 3-mm (ID) test tube, and add
`calcium oxide to a height of 1 cm. Heat the tube in a flame, starting
`from the top, until the reaction is complete, then ignite for a short
`time. Transfer the contents to a beaker containing 10 mL of water,
`acidify with nitric acid, and filter. To the filtrate add 0.2 mL of silver
`nitrate solution ( 1 in 60): any opalescence obtained is not more than
`that obtained from a blank solution treated in the same manner.
`Assay-Transfer about 50 mg of Crotarniton, accurately weighed, to
`a 100-mL volumetric flask, add cyclohexane to volume, and mix .
`Transfer 10.0 mL of this solution to a 250-mL volumetric flask, dilute
`with cyclohexane to volume, and mix. Determine the absorbance of
`this solution and of a solution of USP Crotarniton RS in the same
`medium having a known concentration of about 20 µg per mL in I(cid:173)
`cm cells at the wavelength of maximum absorbance at about 242 run,
`with a suitable spectrophotometer, using cyclohexane as the blank.
`Calculate the quantity, in mg, of C13H17NO in the Crotarniton taken by
`the formula:
`
`2.5C(Aul As),
`in which C is the concentration, in µg per mL, of USP Crotarniton RS
`in the Standard solution; and Au and As are the absorbances of the
`assay solution and the Standard solution, respectively.
`
`Crotamiton Cream
`
`» Crotamiton Cream contains not less than 93.0 percent
`and not more than 107.0 percent of the labeled amount of
`C13H11NO.
`Packaging and storage-Preserve in collapsible tubes or tight, light(cid:173)
`resistant containers.
`USP Reference standards ( 11 )-USP Crotamiton RS.
`Identification-The retention time of the major peak in the
`chromatogram of the Assay preparation corresponds to that of the
`Standard preparation, both relative to the internal standard, as
`obtained in the Assay preparation.
`Minimum fill (755) : meets the requirements.
`Assay-
`Internal standard solution-Dissolve butyl benzoate in methanol
`to obtain a solution containing about 17 .5 mg per mL.
`Mobile phase-Prepare a suitable degassed and filtered mixture of
`acetonitrile and water (3 : 2).
`Standard solution-Dissolve a suitable quantity of USP Crotarni(cid:173)
`ton RS, accurately weighed, in methanol to obtain a solution having a
`known concentration of about 1 mg per mL.
`Standard preparation-Pipet IO mL of Standard solution and 5
`mL of Internal standard solution into a 50-mL volumetric flask,
`dilute with methanol to volume, and mix.
`Assay preparation-Transfer an accurately weighed portion of
`Crotamiton Cream, equivalent to about 50 mg of crotamiton, to a
`tared 50-mL volumetric flask. Add about 25 mL of methanol, and
`shake and sonicate to disperse the cream. Dilute with methanol to
`volume, and mix. Filter about 20 mL through moderately retentive
`filter paper. Pipet 10 mL of the clear filtrate and 5 mL of Internal
`standard solution into a 50-mL volumetric flask, dilute with methanol
`to volume, and mix.
`Procedure-Inject equal volumes of the Standard preparation and
`the Assay preparation into a liquid chromatograph (see Chromatog(cid:173)
`raphy (621)) equipped with a 254-nm detector and a 4.6-mm x 25-
`cm stainless steel column that contains packing LI. In a suitable
`
`Assay preparation-Transfer 4 mL of Nasal Solution to a 100-mL
`volumetric flask, dilute with water to volume, and mix. Transfer an
`aliquot of this solution, equivalent to 8 mg of cromolyn sodium, to a
`250-mL volumetric flask. Add 2.5 mL of pH 7.4 Sodium phosphate
`buffer, dilute with water to volume, and mix.
`Standard preparation-Prepare as directed in the Assay under
`crornolyn Sodium.
`p,ocedu,._Proceed as directed for Procedure in the Assay under
`crornolyn Sodium Inhalation Solution.
`
`Cromolyn Sodium Ophthalmic Solution
`
`» Cromolyn Sodium Ophthalmic Solution is a sterile,
`aqueous solution of Cromolyn Sodium. It contains not
`less than 90.0 percent and not more than 110.0 percent of
`the labeled amount of C23H1~a2011. It may contain
`suitable antimicrobial and stabilizing agents.
`Packaging and storage-Preserve in tight, light-resistant, single(cid:173)
`dose or multiple-dose containers. Ophthalmic Solution that is
`packaged in multiple-dose containers contains a suitable antimicro(cid:173)
`bial agent.
`USP Reference standards (11)-USP Cromolyn Sodium RS.
`Identification-It meets the requirements for Identification test B
`under Cromolyn Sodium.
`Sterility (71 ): meets the requirements.
`pH (791 }: between 4.0 and 7.0.
`Related compounds-It meets the requirements of the test for
`Related compounds under Cromolyn Sodium Inhalation Solution,
`"Ophthalmic Solution" being read in place of "Inhalation Solution."
`Assay-
`•. pH 7.4 Sodium phosphate buffer-Prepare as directed in the Assay
`under Cromolyn Sodium.
`Assay preparation-Transfer 4 mL of Ophthalmic Solution to a
`100-mL volumetric flask, dilute with water to volume, and mix.
`Transfer an aliquot of this solution, equivalent to 8 mg of cromolyn
`sodium, to a 250-mL volumetric flask. Add 2.5 mL of pH 7. 4 Sodium
`phosphate buffer, dilute with water to volume, and mix.
`Standard preparation-Prepare as directed in the Assay under
`Cromolyn Sodium.
`Procedure-Proceed as directed for Procedure in the Assay under
`Cromolyn Sodium Inhalation Solution.
`
`Crotamiton
`
`C"H.,NO 203.28
`tiutenamide, N-ethyl-N-(2-methylphenyl)-.
`[ 483-63-6].
`• thyl-o-crotonotoluidide
`
`» Crotamiton is a mixture of cis and trans isomers
`ntaining not less than 97.0 percent and not more than
`3.0 percent of C13H17NO.
`~ackaging and stonge--Preserve in tight, light-resistant containers.
`SP Reference standards ( 11 )-USP Crotamiton RS.
`
`o0
`c1
`
`omolyn Sodium RS.
`'.llm of the Assay prepara.
`,its maxima and minima at
`;olution of USP CromolYn
`
`:ts the requirements.
`
`. ons of Inhalation Solution
`Sodium RS in a mixture of
`acetone ( 6 : 4 : I) contain(cid:173)
`A) and 0.1 mg per ml
`.yer chromatographic plate
`•ith a 0.25-mm layer of
`-:,w the spots to dry, and
`tern consisting ofa mixture
`tic acid (9 : 9 : 2) until the
`1s of the length of the plate.
`:hamber, mark the solvent
`. ocate the spots on the plate
`light: the RF value of the
`,n Solution corresponds to
`,n A. Any spot in the
`on Solution moving ahead
`nse than the spot in the
`,[ution B (1.0%).
`
`i Standard preparation(cid:173)
`-omolyn Sodium.
`r an accurately measured
`lent to about 25 mg of
`having a concentration of
`1is solution into a 100-ml
`um phosphate buffer, dilute
`
`the absorbances of the
`paration in I -cm cells at
`, at about 326 nm, with a
`00 aqueous solution of pH
`:. Calculate the quantity, in
`1halation Solution taken by
`
`,er mL, of USP Cromolyn
`Vis the volume, in mL, of
`are the absorbances of the
`•aration and the Standard
`
`Solution
`
`lution is an aqueous
`contains not less than
`110.0 percent of the
`Ct may contain suitable
`
`1t, light-resistant containers.
`~romolyn Sodium RS.
`,ts for Identification test B
`
`1uirements of the test _£or
`,dium Inhalation Solunon.
`"Inhalation Solution."
`
`,are as directed in the Ass!J'j
`
`Slayback Exhibit 1055, Page 21 of 78
`Slayback v. Eye Therapies - IPR2022-00142
`
`

`

`USP28
`
`VSP28
`
`Official Monographs I Cyclopentolate
`
`557
`
`Uniformity of dosage units (905): meet the requirements.
`,usay-
`Mobi/e phase-Prepare a suitable filtered and degassed mixture of
`water acetonitrile, methanol, and methanesulfonic acid
`(48 : 28 : 24: 0.2), and adjust with diethylamine to a pH of 3.6.
`Make adjustments if necessary (see System Suitability under
`Chromatography (621}).
`Standard preparation-Dissolve an accurately weighed quantity of
`USP Cyclobenzal(rin~ Hydrochloride J?-8 ~ 0.1 N hydrochl~ric acid,
`and dilute quant1tatively, and stepwise 1f necessary, with 0.1 N
`hydrochloric acid to obtain a solution having a known concentration
`of about 0.05 mg per mL.
`Assay preparation-Weigh and finely powder not fewer than 20
`Tablets. Transfer an accurately weighed portion of the powder,
`equivalent to about 10 mg of cyclobenzaprine hydrochloride, to a
`200-mL volumetric flask, add 150 mL ofO. l N hydrochloric acid, and
`shake by mechanical means for 30 minutes. Dilute with 0.1 N
`hydrochloric acid to volume, mix, and filter.
`Chromatographic system (see Chromatography (621}}-The
`liquid chromatograph is equipped with a 290-nm detector and a
`4.6-mm x 10-cm column that contains packing LI. The flow rate is
`about 1.5 mL per minute. Chromatograph the Standard preparation,
`and record the peak responses as directed for Procedure: the capacity
`factor, K, for the analyte peak is not less than 2.0; the column
`efficiency determined from the analyte peak is not less than 1000
`theoretical plates; the tailing factor for the analyte peak is not more
`than 2; and the relative standard deviation for replicate injections is
`not more than 2.0%.
`Procedure-Separately inject equal volumes (about 10 µL) of the
`Standard preparation and the Assay preparation into the chromat(cid:173)
`ograph, record the chromatograms, and measure the areas for the
`major peaks. Calculate the quantity, in mg, of C,oH,.N • HCI in the
`portion of Tablets taken by the formula:
`
`200C(rulrs),
`in which C is the concentration, in mg per mL, of USP
`Cyclobenzaprine Hydrochloride RS in the Standard preparation:
`and ru and rs are the peak responses obtained from the Assay
`preparation and the Standard preparation, respectively.
`
`Cyclopentolate Hydrochloride
`
`Residue on ignition (281): not more than 0.05% .
`Chromatographic purity-
`Buffer solution, Mobile phase, and Chromatographic system(cid:173)
`Prepare as directed under Assay.
`Test preparation-Use the Assay preparation.
`Procedure-Inject a volume (about 20 µL) of the Test preparation
`into the chromatograph, record the chromatogram obtained for a
`period of not less than twice the retention time of cyclopentolate, and
`measure the peak responses. Calculate the percentage of each peak,
`other than the solvent peak and the cyclopentolate peak, in the
`specimen of Cyclopentolate Hydrochloride taken by the same
`formula:
`
`I00r;lr0
`in which r, is the response of each peak and r, is the sum of the
`responses of all of the peaks, excluding that of the solvent peak: not
`more than 1.0% individual impurity and not more than 2.0% total
`impurities are found.
`Assay-
`Buffer solution-Dissolve 660 mg of dibasic ammonium phos(cid:173)
`phate in 1000 mL of water. Adjust with phosphoric acid to a pH of
`3.0 ± 0.1, and mix.
`Mobile phase-Prepare a suitable filtered and degassed mixture of
`acetonitrile and Buffer solution (1 : 3). Make adjustments ifnecessary
`(see System Suitability under Chromatography (621)).
`Standard preparation-Dissolve an accurately weighed quantity of
`USP Cyclopentolate Hydrochloride RS in water, dilute quantitatively,
`and stepwise if necessary, with water, and mix to obtain a solution
`having a known concentration of about 0.1 mg per mL.
`Assay preparation-Transfer about I 00 mg of Cyclopentolate
`Hydrochloride, accurately weighed, to a 100-mL volumetric flask,
`dilute with water to volume, and mix. Transfer 5.0 mL of this solution
`to a 50-mL volumetric flask, dilute with water to volume, and mix.
`Chromatographic system (see Chromatography (621}}-The
`liquid chromatograph is equipped with a 220-nm detector and a
`4.6-mm x 15-cm column that contains packing LIS. The flow rate is
`about 2 mL per minute. Chromatograph the Standard preparation,
`and record the peak responses as directed for Procedure: the column
`efficiency determined from the analyte peak is not less than 3000
`theoretical plates, the tailing factor for the analyte peak is not more
`than 2.0, and the relative standard deviation for replicate injections is
`not more than 2.0%.
`Procedure-Separately inject equal volumes (about 20 µL) of the
`Standard preparation and the Assay preparation into the chromat(cid:173)
`ograph, record the chromatograms, and measure the responses for the
`major peaks. Calculate the quantity, in mg, of C17HuNO, • HCl in the
`portion of Cyclopentolate Hydrochloride taken by the formula:
`I 000C(rul rs),
`in which C is the concentration, in mg per mL, of USP Cyclopentolate
`Hydrochloride RS in the Standard preparation; and ru and rs are the
`cyclopentolate peak responses obtained from the Assay preparation
`and the Standard preparation, respectively.
`
`C,,H21NO, · HCI 327.85
`Benzeneacetic acid, a-( 1-hydroxycyclopentyl)-, 2-( dirnethylamino )·
`_ethyl ester, hydrochloride, ( ± )·.
`2-(Dimethylamino )ethyl ( ± )-1 -hydroxy-a-phenylcyclopentaneace-
`tate hydrochloride
`[5870-29-1].
`
`Cyclopentolate Hydrochloride
`Ophthalmic Solution
`
`» Cyclopentolate Hydrochloride Ophthalmic Solution is
`a sterile, aqueous solution ofCyclopentolate Hydrochlo(cid:173)
`ride. It may contain suitable buffers and other additives.
`It contains not less than 90.0 percent and not more than
`110.0 percent of the labeled amount ofC17H25N03 • HCI.
`Packaging and storage-Preserve in tight containers, and store at
`controlled room temperature.
`USP Reference standards ( I I }-USP Cyclopentolate Hydrochlo(cid:173)
`ride RS.
`Identification-Place in a 125-mL separator a volume of Ophthalmic
`Solution, equivalent to about 50 mg of cyclopentolate hydrochloride,
`and place in a second separator about 50 mg of USP Cyclopentolate
`Hydrochloride RS dissolved in 5 mL of water. Treat each solution as
`
`>► Cyclopentolate Hydrochloride contains not less than
`98.0 percent and not more than 102.0 percent of
`C11H2sN03 • HCl, calculated on the dried basis.
`kaging and storage-Preserve in tight containers, and store in a
`Pa
`cod place.
`~d,SP Reference standards (11}-USP Cyclopentolate Hydrochlo(cid:173)
`n eRS.
`Identification-
`Infrared Absorption (197K).
`A:
`B: A solution ( I in 500) responds to the tests for Chloride ( 191) .
`pH (791) : between 4.5 and 5.5, in a solution (I in 100).
`~ on drying (731 )-Dry it at I 05° for 4 hours: it loses not more
`.._ 0.5% of its weight.
`
`1c
`
`. 1%.
`
`1n methanol, and dilute
`t solution. Dissolve a
`Hydrochloride RS in
`a known concentration
`solution quantitatively
`1ted standard solution
`•. Apply separate 5-µL
`line of a suitable thin(cid:173)
`graphy (621}) coated
`;ilica gel mixture and
`he chromatogram in a
`nt system consisting of
`, hydroxide (75: 25: 1)
`fourths of the length of
`air-dry, and view under
`principal spot from the
`iard solution; and any
`s not exceed, in size or
`the Diluted standard
`
`) : meets the require-
`
`;ept to use 100.0 µg of
`'6.0 µg of 1,4-dioxane,
`
`zaprine Hydrochloride,
`acetic acid, add 15 ml
`N perchloric acid VS,
`using a platinum ring
`rode containing 0.1 N
`:see Titrimetry (541)).
`y necessary correction.
`ralent to 31.19 mg of
`
`oride Tablets
`
`rablets contain not
`1an 110.0percentof
`,rine hydrochloride
`
`losed containers.
`1benzaprine Hydrochlo-
`
`nely powdered Tablets,
`ine hydrochloride, to a
`e, swirl to dissolve, and
`1L, transfer to a suitable
`ivaporate with the aid of
`:ii crystallization occurs.
`her, and air-dry.
`in the chromatogram of
`the chromatogram of the
`iay.
`
`nL.
`
`H2,N · HCI dissolved by
`of maximum absorbance
`the solution under test,
`if necessary, in compat·
`vn concentration of USP
`ameMedium.
`the labeled amount of
`
`Slayback Exhibit 1055, Page 22 of 78
`Slayback v. Eye Therapies - IPR2022-00142
`
`

`

`558
`
`Cyclophosphamide / Official Monographs
`
`USP28
`
`USP28
`
`follows. Add 1 g of potassium carbonate, and extract with two 10-mL
`portions of ether. Pass the ether extracts through ether-washed filter
`paper, collect the filtrate in a small beaker, and evaporate to dryness:
`the residue so obtained responds to Identification test A under
`Cyclopentolate Hydrochloride.
`Sterility (71 ): meets the requirements.
`pH (791 ): between 3.0 and 5.5.
`Assay-
`Buffer solution, Mobile phase, Standard preparation, and
`Chromatographic system-Proceed as directed in the Assay under
`Cyclopentolate Hydrochloride.
`Assay preparation-Transfer an accurately measured volume of
`Ophthalmic Solution, equivalent to about IO mg of cyclopentolate
`hydrochloride, to a I 00-mL volumetric flask, dilute with water to
`volume, and mix.
`Procedure-Proceed as directed in the Assay under Cyclopentolate
`Hydrochloride. Calculate the quantity, in mg, of cyclopentolate
`hydrochloride (C 17H2,NO, · HCI) in each mL of the Ophthalmic
`Solution taken by the formula:
`
`IOO(C I V)(ru l r.),
`in which Vis the volume, in mL, of Ophthalmic Solution taken, and
`the other terms are as defined therein.
`
`Cyclophosphamide
`
`mL of Internal standard solution, dilute with water to volume, and
`mix to obtain a Standard preparation having a known concentration
`of about 0.5 mg of anhydrous cyclophosphamide per mL.
`Assay preparation-Transfer an accurately weighed quantity of
`Cyclophosphamide, equivalent to about 200 mg of anhydrous
`cyclophosphamide, to a 200-mL volumetric flask, add about 50 mL
`of water, shake for about 5 minutes, dilute with water to volume, and
`mix. Pipet 25 mL of this solution and 5 mL of Internal standard
`solutio_n into a 50-mL volumetric flask, dilute with water to volume,
`and mtx.
`Chromatographic system (see Chromatography {621)}-The
`liquid chromatograph is equipped with a 195-nm detector and a
`3.9-mm x 30-cm column that contains packing LI. The flow rate is
`about 1.5 mL per minute. Chromatograph six replicate injections of
`the Standard preparation, and record the peak responses as directed
`for Procedure: the relative standard deviation is not more than 2%,
`and the resolution factor between cyclophosphamide and ethylpar(cid:173)
`aben is not less than 2.
`Procedure-Separately inject equal volumes (about 25 µL) of the
`Standard preparation and the Assay preparation into the chromat(cid:173)
`ograph, record the chromatograms, and measure the responses for the
`major peaks. The relative retention times are about 0.7 for
`cyclophosphamide and 1.0 for ethylparaben. Calculate the quantity,
`in mg, of C,H.,CI2N2O,P in the Cyclophosphamide taken by the
`formula:
`
`400C(Rul Rs),
`in which C is the concentration, in mg per mL, of anhydrous
`cyclophosphamide in the Standard preparation, as determined from
`the concentration of USP Cyclophosphamide RS corrected for
`moisture content by a titrimetric water determination; and Ru and Rs
`are the ratios of the peak responses of cyclophosphamide to those of
`ethylparaben in the Assay preparation and the Standard preparation,
`respectively.
`
`C,H,,Cl2N2O2P-H,O 279.10
`2H-l ,3,2-Oxazaphosphorin-2-amine, N,N-bis(2-chloroethyl)tetrahy(cid:173)
`dro-, 2-oxide, monohydrate, ( ± ).
`( ± )-2-[Bis(2-chloroethyl)amino )tetrahydro-2H-l ,3,2-oxazaphos-
`[6055-19-2].
`phorine 2-oxide monohydrate
`[50-18-0].
`Anhydrous
`261.09
`
`» Cyclophosphamide contains not less than 97 .0 percent
`and not more than 103.0 percent of C1H1sCliN2O2P,
`calculated on the anhydrous basis.
`Caution-Great care should be taken in handling
`Cyc/ophosphamide, as it is a potent cytotoxic agent.
`
`Packaging and storage-Preserve in tight containers, at a
`temperature between 2° and 30°.
`USP Reference standards ( 11 )-USP Cyclophosphamide RS.
`Identification-
`A:
`Infrared Absorption (197K).
`B: The retention time of the major peak in the chromatogram of
`the Assay preparation corresponds to that of the Standard
`preparation, both relative to the internal standard, as obtained in
`the Assay.
`pH (791) : between 3.9 and 7.1, in a solution (I in 100), determined
`30 minutes after its preparation.
`Water, Method l (921) : between 5.7% and 6.8%.
`Heavy metaJs {231)-Dissolve 1.0 gin 25 mL of water, and filter if
`necessary: the limit is 0.002%.
`Assay-
`Mobile phase-Prepare a suitable, degassed solution of water and
`acetonitrile (70 : 30).
`Internal standard solution-Dissolve about I 85 mg of ethylpar(cid:173)
`aben in 250 mL of alcohol in a I 000-mL volumetric flask, dilute with
`water to volume, and mix.
`Standard preparation-Transfer an accurately weighed quantity of
`USP Cyclophosphamide RS, equivalent to about 25 mg of anhydrous
`cyclophosphamide, to a 50-mL volumetric flask, add about 25 mL of
`water, and shake to dissolve the USP Reference Standard. Add 5.0
`
`Cyclophosphamide for Injection
`
`» Cyclophosphamide for Injection is a sterile mixture of
`Cyclophosphamide with or without a suitable diluent. It
`contains not less than 90.0 percent and not more than
`110.0 percent of the labeled amount of anhydrous
`cyclophosphamide (C1H1sCliN2O2P).
`Packaging and storage-Preserve in Containers for Sterile Solids as
`described under Injections (I). Storage at a temperature not
`exceeding 25° is recommended. It will withstand brief exposure to
`temperatures up to 30°, but is to be protected from temperatureS
`above 30°.
`USP Reference standards ( 11 )-USP Cyclophosphamide RS. USP
`Endotoxin RS.
`Constituted solution-At the time of use, it meets the requirements
`for Constituted Solutions under Injections (I) .
`Identification-
`A:
`It responds to the Thin-layer Chromatographic Identification
`Test (201), a solution of it in chloroform, equivalent to 20 mg of
`cyclophosphamide per mL, filtered if necessary, being used as the teSI
`solution. Apply 5 µL of the test solution and the Standard solution,
`use a solvent system consisting of a mixture of chloroform, methano~
`and ammonium hydroxide (75 : 20: 5), and visualize the spots by
`placing the plate in an iodine chamber.
`B: The retention time of the major peak in the chromatogram of
`the Assay preparation corresponds to that of the Standar!1
`preparation, both relative to the internal standard, as obtained Ill
`the Assay.
`Bacterial endotoxins (85)-It contains not rriore than 0.20 USP
`Endotoxin Unit per mg of cyclophosphamide.
`pH (791) : between 3.0 and 9.0, but the range does not exceed 3 pH
`units, in a solution containing the equivalent of 20 mg of anhydro~
`cyclophosphamide per mL, determined 30 minutes after its
`preparation.
`
`Other requirements--1
`(11), Uniformity of D
`Injections (I) .
`Assay-
`Mobile phase, lnterna
`/ion-Prepare as directed
`Assay preparation-A
`mide for Injection, eqt
`cyclophosphamide, and I
`the Assay under Cycloph
`Chromatographic sys.
`graphic system in the As:
`Procedure-Proceed a:
`Cyclophosphamide. Cale•
`in the portion of Cycle
`fonnula:
`
`in which the terms are as
`
`Cyclophospharr
`
`» Cyclophosphamide
`percent and not more
`amount of anh·
`(C1H1sCl2N2O2P).
`.
`
`Packaging and storage-(cid:173)
`temperature not exceedi
`withstand brief exposure
`protected from temperaturi
`USP Reference standard:
`Identification-
`A: Extract a portion c
`about 50 mg of cyclophos1
`about 2 mL of the chlorofo1
`potassium bromide, evapor
`last trace of solvent in a s1
`prepare a potassium bromi,
`of the potassium bromide
`between 6.5 and 14 µm, o
`similar preparation of USP
`B: The retention time 1
`the Assay preparation corre
`Standard preparation, as o
`Disintegration {70 I): 3C
`Uncoated Tablets.
`Uniformity of dosage unit
`Procedure for content un
`Perchloric acid solution(cid:173)
`Water, and dilute with wate:
`4-(p-Nitrobenzyl)pyridint
`benzyJ)pyridine in 200 mL
`Sodium hydroxide solutio
`1000 mL of diluted alcohol
`Procedure-Place I Tab!<
`that the final concentration
`about two-thirds full of wa
`disintegrated, dilute with w:
`first 10 mL of the filtrate. P
`tubes 2.0 mL of the filtrate,
`2.0 mL of the Standard solut
`V.:eighed quantity of U