USP
`
`Hydroxyamphetamine
`
`um molybdate in 10 rnLor ·
`,out 2 mg of Hydroxyain,
`blue color is produCQI
`1ds such as amphetarn·
`phenolic hydroxyl, do :: ·
`
`.vater, and add a solution of .
`water. ~xtract with two IO.
`::r solution to evaporate to •
`yamphetamine so obtained
`I under Melting Range o;
`
`~ 10 mL of water add I llli. •
`~: a pale _yellow precipitate
`ammomum hydroxide.
`j 192°.
`· 2 hours: it loses not mort .
`
`an0. 1%.
`ut 400 mg, and dissolve in
`md IO inL of glacial acetic
`th O .I N silver nitrate VS.
`ent to 7.990 mg of Br: the
`;is, is between 33.6% and
`
`c;bloric aci~ to 25 mL, and proceed as d!rected ~mder ldentification--:(cid:173)
`o,ganic Nitrogenous Bases (181}, using sodium carbonate TS m
`p(ace of I N sodium hydroxide, beginning with "Transfer the liquid
`to a separator": the Ophthalmic Solution meets the requirements of
`merest.
`sterility (7 I}: meets the requirements.
`pff (791} : between 4.2 and 6.0.
`,usay-Trans~er an accurately measured volume of Ophthalmic
`5olution, equivalent to about 100 mg of hydroxyamphetamine
`bydrobromide, to a 125-mL separator. Wash the solution with 15 mL
`of chloroform, and discard the washing. Rinse the stopper and the
`IIIOllth of_ the separator with_ a ~ew drops of w~ter. Add l.~5 g of
`sodium bicarbonate, preventmg 1t from commg m contact with the
`1110uth of the separator, and swirl until most of the bicarbonate has
`dissolved. By ~eans_ of a l~mL syringe, rapidly inject 0.5 mL of
`acetic anhydnde directly mto the contents of the separator.
`munediately insert the stopper in the separator, and shake vigorously
`until the evolution of carbon dioxide has ceased (7 to 10 minutes),
`releasing the pressure as necessary through the stopcock. Allow to
`stand for 5 minutes, and extract the solution with five I 0-rnL portions
`of chloroform, filtering each extract through a pledget of cotton,
`previously washed with chloroform, into a tared 100-mL beaker.
`Evaporate the combined chloroform extracts on a steam bath in a
`current of air or stream of nitrogen to dryness. Dry the residue at 80°
`for 90 minutes, cool in a desiccator, and weigh. The weight of the
`diacetylhydroxyamphetamine so obtained, multiplied by 0.9866,
`represents the weight of C.HuNO • HBr in the volume of Ophthalmic
`Solution taken.
`
`,thanol, and
`
`Hydroxychloroquine Sulfate
`
`:yamphetamine Hydrobro-
`0 mL of glacial acetic acid
`1g slightly, if necessary, to
`itrate with 0. 1 N perchloric
`and make any necessary
`icid is equivalent to 23.21
`
`,drobromide
`
`,romide Ophthalmic
`aqueous solution of
`1ide. It contains not
`than 105.0 percent of
`· HBr. It contains a
`
`, light-resistant containers.
`"iydroxyamphetamine HJ"
`
`1m molybdate in IO mL of
`c Solution: an intense blue
`amino compounds such OS
`~ich, lacking a phenolic
`
`nine obtained in the Assay
`· under Melting Range orf
`en beginning and end o
`
`D under Hydroxyamphel·
`
`C,.H,.CIN30 · H,SO, 433.95
`Ethanol, 2-(( 4-((7-chloro-4-quinolinyl)amino )pentyl]ethyl]amino-,
`(±)-, sulfate (I : I) (salt).
`( ± )-2-[[ 4-((7-Chloro-4-quinolyl)amino ]pentyl]ethylamino ]ethanol
`[747-36-4].
`sulfate (I : I) (salt)
`
`» Hydroxychloroquine Sulfate contains not less than
`98.0 percent and not more than 102.0 percent of
`C1sH26CIN3O · H2SO4, calculated on the dried basis.
`Pac~ging and storage-Preserve in well-closed, light-resistant
`containers.
`USP Reference standards ( 11 )-USP Hydroxychloroquine Sulfate
`RS.
`Identification-
`A: Ultraviolet Absorption (197U)(cid:173)
`Solution: 10 µg per mL.
`Medium: dilute hydrochloric acid (I in 100).
`B:
`Infrared Absorption (197K).
`C: A solution ( I in I 00) responds to the tests for Sulfate ( I 9 I}.
`loss on drying (731 }-Dry it at I 05° for 2 hours: it loses not more
`tha
`n 2.00/o of its weight.
`Ordinary impurities (466}-
`Test solution: 10% water in methanol.
`Standard solution: 10% water in methanol.
`Eluant:
`a mixture of alcohol, water, and ammonium hydroxide
`(80
`: 16 :4).
`Visualization:
`1.
`Organic volatile impurities, Method I (467): meets the require(cid:173)
`lllents.
`Assay-Dissolve about I 00 mg of Hydroxychloroquine Sulfate,
`:urately _weighed, in about 5 mL of water, and dilute quantitatively
`stepwise with dilute hydrochloric acid ( I in I 00) to obtain a
`
`Official Monographs I Hydroxychloroquine
`
`977
`
`solution containing about IO µg per mL. Similarly prepare a Standard
`solution of USP Hydroxychloroquine Sulfate RS. Concomitantly
`determine the absorbances of both solutions in I-cm cells at the
`wavelength of maximum absorbance at about 343 nm, with a suitable
`spectrophotometer, using dilute hydrochloric acid ( I in I 00) as the
`blank. Calculate the quantity, in mg, of C 11H,0CIN30 • H,SO, in the
`portion of Hydroxychloroquine Sulfate taken by the formula:
`
`I 0C(Aul As),
`
`in which C is the concentration, in µg per mL, of USP
`Hydroxychloroquine Sulfate RS in the Standard solution, and Au
`and As are the absorbances of the solution of Hydroxychloroquine
`Sulfate and the Standard solution, respectively.
`
`Hydroxychloroquine Sulfate Tablets
`
`» Hydroxychloroquine Sulfate Tablets contain not less
`than 93.0 percent and not more than 107.0 percent of the
`labeled amount of hydroxychloroquine sulfate
`(C1sH26CIN3O · H2SO4).
`
`Packaging and storage-Prese1ve in tight, light-resistant containers.
`USP Reference standards ( 11 )-USP Hydroxychloroquine Sulfate
`RS.
`Identification-
`A: Triturate a quantity offinely powdered Tablets, equivalent to
`about I g of hydroxychloroquine sulfate, with 50 mL of water, and
`filter: the clear filtrate so obtained meets the requirements for
`Identification tests B and C.
`B:
`It meets the requirements under Identification- Organic
`Nitrogenous Bases ( 181 ).
`C: A solution (I in I 00) meets the requirements of the tests for
`Sulfate (1 91 ).
`D:
`the retention time of the major peak in the chromatogran1 of
`the Assay preparation corresponds to that in the chromatogran1 of the
`Standard preparation, as obtained in the Assay.
`Dissolution (711 }-
`Medium: water; 900 mL.
`Apparatus 2: 50 rpm.
`Time: 60 minutes.
`Procedure- Determine the amount of C 18H26CIN,O · H,SO, dis(cid:173)
`solved from UV absorbances at the wavelength of maximum
`absorbance at about 343 nm of 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 Hydroxychloroquine Sulfate RS in the same medium.
`Tolerances- Not less than 70% (Q) of the labeled amount of
`C,.H;i.CINiO • H,SO, is dissolved in 60 minutes.
`Uniformity of dosage units (905) : meet the requirements.
`Assay-
`Mobile phase-To 800 mL of water, add I 00 mL of methanol, I 00
`mL ofacetonitrile, 2.0 ml of phosphoric acid, and 96 mg of sodium
`1-pentanesulfonate, mix, and filter. Make adjustments if necessary
`(see System Suitability under Chromatography (621)).
`Solvent mixture-Prepare a mixture of methanol and water (I : !).
`Standard preparation-Dissolve an accurately weighed quantity of
`USP Hydroxychloroquine Sulfate RS in Solvent mixture, dilute
`quantitatively with Solvent mixture, and mix to obtain Solution A
`having a known concentration of about I mg per mL. Transfer 5.0 mL
`of this solution to a I 00-mL volumetric flask, dilute with Mobile
`phase to volume, and mix to obtain the Standard preparation having
`a known concentration of about 0.05 mg per mL.
`Resolution solution-Prepare a solution of chloroquine phosphate
`in methanol having a concentration of I mg per mL. Transfer 5.0 mL
`of this solution to a 100-mL volumetric flask, add 5.0 mL of Solution
`A, dilute with Mobile phase to volume, and mix.
`Assay preparation-Weigh and finely powder not less than 20
`Tablets. Transfer an accurately weighed portion of the powder,
`equivalent to about 200 mg of hydroxychloroquine sulfate to a 200-
`mL volumetric flask, add about I SO mL of Solvent mixture, and mix.
`Sonicate, with intermittent shaking, for about I 5 minutes, and cool to
`
`Slayback Exhibit 1055, Page 40 of 78
`Slayback v. Eye Therapies - IPR2022-00142
`
`

`

`06.28
`·c acid, a-n
`tylhydratroJ
`sobutylphen)
`
`[58560
`
`tification-
`• Infrared Absorpti<
`B: Ultraviolet Abso
`lution: 250 µg pe
`edium: 0.1 N sodi
`·Respective absm:ptivi
`drous basis, do not
`'J:: The chromatogr
`in the Assay
`·on time of whicl
`nds to that exh.
`tion, obtained Ill
`, Method I (921):
`ue on ignition (2!
`metals, Method
`. matograpbic puri
`, . bile phase-Prep:
`ously adjusted wi
`itrile (1340: 680).
`ility under Chrom,
`t preparation-Pre
`· ning about 5 mg J:
`esolution solution-:
`each ml about 5 mg
`. matographic sy.
`chromatograph is
`. x 15-cm column
`"ned at 30 + 0.2°
`. . ograph a series ,
`on the column. C
`. the peak respom
`n times are about
`the resolution, R,
`fen peak is not les
`edure-(NOTE-U
`. ] Inject about
`tograph, record ·
`es. Calculate the
`la:
`
`, is the respc:
`and the ma
`all the peal
`.3% of any i1
`idual impuri1
`olatile imp
`
`Use dimethy
`butylaceto
`paration and th
`
`of hydroxypropoxy (-OCH,CHOHCH,) in the Hypromellose
`by the formula:
`
`2(75 / l 70)F,Ru,(W7/ Wu),
`in which 75/l"~0 is the _ra~o of ~e formula weights of hydro!
`xypropoxy and 1sopropyl 1od1de; F, 1s defined under Calibration· J
`is the ratio of the area of the isopropyl iodide peak to that of
`toluene peak obtained from the Assay preparation; W7 is the w~
`in g, of toluene in the Internal standard solution; and Wu is !lit
`.1,
`weight, in g, of Hypromellose taken for the Assay.
`
`Hypromellose Ophthalmic Solution
`
`» Hypromellose Ophthalmic Solution is a sterile solu · ·
`of Hypromellose. It contains not less than 85.0
`and not more than 115.0 percent of the labeled amount
`Hypromellose (hydroxypropyl methylcellulose). It
`·
`contain suitable antimicrobial, buffering, and stabil' ·
`agents.
`
`Packaging and storage-Preserve in tight containers.
`USP Reference standards ( 11 }-USP Hydroxypropyl Methyl
`lose RS.
`Identification-
`It meets the requirements of Identification test C
`A:
`Hypromellose.
`B: Heat 5 ml of Ophthalmic Solution in a test tube over a
`flame: the warm solution turns cloudy but clears upon chilling . . :
`Sterility (71): meets the requirements.
`pH (791): between 6.0 and 7.8.
`Assay-
`Standard preparation-Dissolve a suitable quantity of
`Hydroxypropyl Methylcellulose RS, accurately weighed, in
`and dilute quantitatively with water to obtain a solution ha ·
`known concentration of about l 00 µg per ml.
`Assay preparation-Dilute an accurately measured volume.
`Ophthalmic Solution quantitatively with water to obtain a so
`having an equivalent concentration of about I 00 µg of hyprom
`per ml.
`Procedure-Pipet 2 ml each of the Standard preparation,
`Assay preparation, and water to provide a blank, into separate,
`stoppered test tubes. To each tube add 5.0 ml of dipheny
`solution (prepared by dissolving 3.75 g of colorless dipheny
`crystals in 150 ml of glacial acetic acid and diluting the solution
`90 ml of hydrochloric acid), mix, and immediately insert the
`into an oil bath at I 05° to 110° for 30 minutes, the temperature
`kept uniform within 0.1 ° during heating. Remove the tubes, and P
`them in an ice-water bath for IO minutes or until thoroughly
`·
`room temperature and using a suitable spectrophotometer, con
`itantly determine the absorbances of the solutions from the St
`preparation and the Assay preparation at 635 nm, using the
`solution as the blank. Calculate the quantity, in mg, ofhyprom
`in each ml of the Ophthalmic Solution taken by the fonnula:
`
`0.001 C(d I V)(Aul As),
`in which C is the concentration, in µg per ml, of USP Hydrox
`Methylcellulose RS in the Standard preparation; Vis the vo
`ml, of Ophthalmic Solution taken; d is the dilution fold of V cd
`obtain the Assay preparation; and Au and As are the absorban
`the solutions from the Assay preparation and the Sta
`preparation, respectively.
`
`JIII:!
`
`1U
`
`990
`
`Hypromellose / Official Monographs
`
`Heavy metals, Method II (231) : 0.001%, l ml ofhydroxylamine
`hydrochloride solution (l in 5) being added to the solution of the
`residue.
`Organic volatile impurities, Method IV {467): meets the require(cid:173)
`ments.
`Assay-{Caution-Hydriodic acid and its reaction byproducts are
`highly toxic. Perform all steps of the Assay preparation and the
`Standard preparation in a properly functioning hood. Specific safety
`practices to be followed are to be identified to the analyst performing
`this test.]
`Hydriodic acid-Use a reagent having a specific gravity of at least
`1.69, equivalent to 55% HI.
`Internal standard solution-Transfer about 2.5 g of toluene,
`accurately weighed, to a I 00-ml volumetric flask containing l 0
`ml of o-xylene, dilute with o-xylene to volume, and mix.
`Standard preparation-Into a suitable serum vial weigh about 135
`mg of adipic acid and 4.0 ml of Hydriodic acid, pipet 4 ml of
`Internal standard solution into the vial, and close the vial securely
`with a suitable septum stopper. Weigh the vial and contents
`accurately, add 30 µL of isopropyl iodide through the septum with
`a syringe, again weigh, and calculate the weight of isopropyl iodide
`added, by difference. Add 90 µL of methyl iodide similarly, again
`weigh, and calculate the weight of methyl iodide added, by
`difference. Shake, and allow the layers to separate.
`Assay preparation-Transfer about 0.065 g of dried Hypromellose,
`accurately weighed, to a 5-ml thick-walled reaction vial equipped
`with a pressure-tight septum-type closure, add an amount of adipic
`acid equal to the weight of the test specimen, and pipet 2 ml of
`Internal standard into the vial. Cautiously pipet 2 ml of Hydriodic
`acid into the mixture, immediately cap the vial tightly, and weigh
`accurately. Mix the contents of the vial continuously, while heating at
`150° for 60 minutes. Allow the vial to cool for about 45 minutes, and
`again weigh. If the weight loss is greater than l 0 mg, discard the
`mixture, and prepare another Assay preparation.
`Chromatographic system (see Chromatography (621))-The gas
`chromatograph is equipped with a thermal conductivity detector and a
`4-mrn x 1.8-m glass column packed with 20% liquid phase G28 on
`100- to 120-mesh support SIC that is not silanized. Helium is used as
`the carrier gas and the temperature of the column is maintained at
`130°. Chromatograph the Standard preparation, and record the peak
`responses as directed for Procedure: the relative retention times are
`about 1.0, 2.2, 3.6, and 8.0 for methyl iodide, isopropyl iodide,
`toluene, and o-xylene, respectively; and the resolution, R, between
`toluene and isopropyl iodide is not less than 2.0.
`Calibration-Inject about 2 µL of the upper layer of the Standard
`preparation into the gas chromatograph, and record the chromato(cid:173)
`gram. Calculate the relative response factor, F .,, of equal weights of
`toluene and methyl iodide taken by the formula:
`
`Q.,!Rs,,,,
`in which Q., is the quantity ratio of methyl iodide to toluene in the
`Standard preparation, and Rs., is the peak area ratio of methyl iodide
`to toluene obtained from the Standard preparation. Similarly,
`calculate the relative response factor, F,, of equal weights of toluene
`and isopropyl iodide taken by the formula:
`Q1/R51,
`in which Q, is the quantity ratio of isopropyl iodide to toluene in the
`Standard preparation, and Rs, is the peak area ratio of isopropyl
`iodide to toluene obtained from the Standard preparation.
`Procedure-Inject about 2 µL of the upper layer of the Assay
`preparation into the gas chromatograph, and record the chromato(cid:173)
`gram. Calculate the percentage of methoxy (-OCH,) in the
`Hypromellose taken by the formula:
`
`2(31 / 142)F..,Ru.,(W7/Wu),
`in which 31/142 is the ratio of the formula weights ofmethoxy and
`methyl iodide; F., is defined under Calibration; Ru., is the ratio of the
`area of the methyl iodide peak to that of the toluene peak obtained
`from the Assay preparation; W7 is the weight, in g, of toluene in the
`Internal standard solution; and Wu is the weight, in g, of
`Hypromellose taken for the Assay. Similarly, calculate the percentage
`
`Slayback Exhibit 1055, Page 41 of 78
`Slayback v. Eye Therapies - IPR2022-00142
`
`

`

`Official Monographs I ldoxuridine
`
`997
`
`Standard preparation-Transfer about 25 mg of USP Idoxuridine
`RS, accurately weighed, to a 50-mL volumetric flask, add methanol
`to volume, and mix. Dilute 5.0 mL of this solution with a mixture of l
`volume of butyl alcohol and 5 volumes of chloroform to l 00.0 mL,
`and mix.
`Assay preparation-Mix 4 g of chromatographic siliceous earth
`with 2 mL of 0.1 N hydrochloric acid in a glass mortar until the
`mixture is fluffy. Add a quantity of Ophthalmic Ointment, equivalent
`to about 5 mg of idoxuridine and accurately weighed, to the mixture,
`and mix.
`Procedure-Transfer the Assay preparation to the prepared
`Chromatographic column . Transfer 2 g of chromatographic siliceous
`earth and 2 mL of0. l N hydrochloric acid to the glass mortar, and mix
`until fluffy, using this material to rinse the mortar and pick up any
`remaining Ophthalmic Ointment. Transfer about half of this mixture
`to the tube, and tamp gently until the column appears uniform.
`Transfer the remaining portion to the Chromatographic column, and
`tamp as before. Wipe the walls of the mortar with a small pledge! of
`glass wool, and insert the pledget in the top of the column. Pass 50
`mL of chloroform through the column at a flow rate of approximately
`l mL per minute, and discard the chloroform. Elute with about 200
`mL of a mixture of I volume of butyl alcohol and 5 volumes of
`chloroform at the same flow rate, discarding the first 20 mL of the
`eluate. Collect the remainder of the eluate in a 200-mL volumetric
`flask, dilute with the eluting solvent to volume, and mix.
`Concomitantly determine the absorbances of this solution and the
`Standard preparation in I-cm cells at 320 nm and at the wavelength
`of maximum absorbance at about 283 nm, with a suitable
`spectrophotometer, using a mixture of butyl alcohol and chloroform
`as the blank. Calculate the quantity, in mg, of C9H 11IN,O, in the
`Ophthalmic Ointment taken by the formula:
`
`0.2C(Am -Am)ul (Am - Am)s,
`in which C is the concentration, in µg per mL, of USP ldoxuridine RS
`in the Standard preparation; and the parenthetic expressions are the
`differences in the absorbances of the two solutions at the wavelengths
`indicated by the subscripts, for the solution from the Ophthalmic
`Ointment (U) and the Standard preparation (S), respectively.
`
`Idoxuridine Ophthalmic Solution
`
`» Idoxuridine Ophthalmic Solution is a sterile, aqueous
`solution of Idoxuridine. It contains not less than 0.09
`percent and not more than 0.11 percent ofC9H11IN2O5• It
`may contain suitable buffers, stabilizers, and antimicro(cid:173)
`bial agents.
`Packaging and storage-Preserve in tight, light-resistant containers
`in a cold place.
`USP Reference standards ( 11 }- USP ldoxuridine RS.
`Identification-The UV absorption spectrum of the solution
`employed for measurement of absorbance in the Assay exhibits
`maxima and minima at the same wavelengths as that of the Standard
`preparation prepared for the Assay.
`Sterility(7 I}: meets the requirements.
`pH (791 }: between 4.5 and 7.0.
`Assay-
`Chromatographic column and Standard preparation-Prepare as
`directed in the Assay under Idoxuridine Ophthalmic Ointment.
`Assay preparation- Mix an accurately measured volume of
`Ophthalmic Solution, equivalent to about 5 mg of idoxuridine, with
`3 g of chromatographic siliceous earth in a glass mortar until the
`mixture is fluffy.
`
`Injection is a s
`,ride and Lactosei
`t1t and not more ·
`1t of C26H21NO9 • H"
`be taken to p
`Hydrochloride
`
`·
`
`.
`
`he Assay preparation .'
`for idarubicin, the re
`the chromatogram of,
`:ay.
`; not more than 8.9
`ydrochloride, a soluti
`1 containing 0.07
`sed in the Test Proc
`. when tested as direc
`,rility of the Product
`
`ution constituted as d'
`!iluent.
`4.0%, the Test Prep
`opic specimen.
`1irements for Uniform ·
`1der Injections (I}.
`
`ntents of l containot'
`quantitatively with
`·.
`mt 0.5 mg of idaru
`
`Procedure under Ida
`n mg, of C,6H2,NO, ·
`ie for Injection taken b~
`
`·ulr5) ,
`.,
`µg per mL, of i~
`Standard prepar~t,o~, ,
`1bicin hydrochlon~e Ill'
`mg per mL, of 1
`1 on the basis of the 1
`of dilution; and ru and r
`; obtained from the
`.
`'on. respectively.
`
`I
`
`...
`J(O
`0)-_N
`HOtJ
`°"
`
`JN,0, 354.10
`1 • 2'~deoxy-5-iodo-.
`;y-5-iodouridine
`
`[54-42-2].
`
`xuridine contains not less than 98.0 percent and not
`than 101.0 percent of C9H11 IN20 5, calculated on
`'dried basis.
`·ng and storage-Preserve in tight, light-resistant containers.
`Reference standards ( 11 }-USP ldoxuridine RS.
`tion-
`lnfrared Absorption (197M}.
`Ultraviolet Absorption ( I 91U)(cid:173)
`tion: 35 µg per mL.
`·um: pH 12.0 buffer (prepared from 7.46g of potassium
`· de and 24 mL of I N sodium hydroxide dissolved in 2000 mL of
`).
`
`rptivities at 279 nm, calculated on the dried basis for the test
`only, do not differ by more than 2.0%.
`· on drying (731 )-Dry about 500 mg, accurately weighed, in
`at 60° for 2 hours: it loses not more than 1.0% of its weight.
`issolve about 250 mg of ldoxuridine, accurately weighed,
`mL of dimethylformamide that previously has been neutralized
`0.1 N sodium methoxide in toluene VS, a solution of 300 mg of
`· I blue in I 00 mL of methanol being used as the indicator.
`with 0.1 N sodium methoxide in toluene VS to a blue
`int, talcing precautions against absorption of atmospheric
`dioxide. Perform a blank determination, and make any
`correction. Each mL of 0.1 N sodium methoxide is
`ent to 35.41 mg ofC9H 11IN,O,.
`
`~_'xuridine Ophthalmic Ointment
`
`. xuridine Ophthalmic Ointment is Idoxuridine in a
`. latum base. It contains not less than 0.45 percent
`~ not more than 0.55 percent of C9H11IN2Os. It is
`
`· g _and storage-Preserve in collapsible ophthalmic oint(cid:173)
`_tubcs m a cool place.
`,Jteference standards ( 11 )-USP ldoxuridine RS.
`- ca~o~ The UV absorption spectrum of the solution from ~e
`ftii exh1b1ts maxima and minima at the same wavelengths as
`tc O_m_tment employed for measurement of absorbance m
`~ e Standard preparation prepared for the Assay.
`(7 I): meets the requirements.
`· Particles-It meets the requirements of the test for Metal
`in Ophthalmic Ointments (751} .
`
`·· . i mL of0. l N hydrochloric acid in a glass mortar until the
`With tographic column-Mix 4 g of chromatographic siliceous
`p,):, uffy. Transfer to a 19- x 250-mm chromatographic tube
`· titte~at~graphy ( 621)) that contains a pledget of glass wool
`. ., with a stopcock at the bottom. Tamp gently to compress
`•0rm mass.
`
`Slayback Exhibit 1055, Page 42 of 78
`Slayback v. Eye Therapies - IPR2022-00142
`
`

`

`998
`
`Ifosfamide / Official Monographs
`
`Procedure-Proceed as directed for Procedure in the Assay under
`ldoxuridine Ophthalmic Ointment, omitting the treatment of the
`column with 50 mL of chloroform. Calculate the quantity, in mg, of
`C9H 11IN,O, in each mL of the Ophthalmic Solution taken by the
`formula:
`
`0.2C(Am - A32o)ulV(Am -A32o)s,
`
`in which C is the concentration, in µg per mL, of USP Idoxuridine RS
`in the Standard preparation; Vis the volume, in mL, of Ophthalmic
`Solution taken; and the parenthetic expressions are the differences in
`the absorbances of the two solutions at the wavelengths indicated by
`the subscripts, for the Solution (U) and the Standard preparation (S),
`respectively.
`
`Ifosfamide
`
`0 H
`
`CO ~,..N.........,.....CI
`
`I
`N ............... CI
`
`C,H,,Cl,N,O,P 261.09
`2H-l ,3,2-0xazaphosphorin-2-amine,N,3-bis(2-chloroethyl)tetrahy(cid:173)
`dro-, 2-oxide.
`3-(2-Chloroethyl)-[(2-chloroethyl)amino ]tetrahydro-2H-l ,3,2-oxaza-
`[3778-73-2].
`phosphorine 2-oxide
`
`» lfosfamide contains not less than 98.0 percent and not
`more than 102.0 percent ofC1H1sChN202P.
`Caution-Great care should be taken in handling
`lfosfamide, as it is a potent cytotoxic agent and
`suspected carcinogen.
`Packaging and storage--Preserve in tight containers at a temper(cid:173)
`ature not exceeding 25 °.
`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 Endotoxin RS. USP lfosfa(cid:173)
`mide 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 4.0 and 7.0 in a solution (I in 10).
`Water, Method I (921): not more than 0.3%.
`Heavy metals, Method I (231): not more than 0.002%.
`Ionic chloride--
`Standard sodium chloride solution-Transfer about 118.7 mg of
`sodium chloride, accurately weighed, to a 200-mL volumetric flask,
`dissolve in and dilute with water to volume, and mix. This solution
`contains 360 ppm of ionic chloride.
`Procedure-Pipet IO mL of Standard sodium chloride solution
`into a beaker, and add 90 mL of water and IO mL of acetic acid.
`Titrate with 0.0 I N silver nitrate VS (prepared fresh daily),
`determining the endpoint potentiometrically using silver and silver(cid:173)
`silver chloride electrodes. Record the volume, V,, of 0.01 N silver
`nitrate VS consumed. Transfer about 2.0 g of lfosfamide, accurately
`weighed, into a beaker, and add 90 mL of water and IO mL of acetic
`acid. Pipet IO mL of Standard sodium chloride solution into the
`beaker, and stir, if necessary, until solution is complete. Titrate with
`0.01 N silver nitrate VS as directed above, and record the volume, V,,
`of 0.01 N silver nitrate VS consumed. Calculate the difference in
`volume, V, of 0.01 N silver nitrate VS consumed between the two
`determinations by subtracting V, from V,: a difference of not more
`than 1.0 mL corresponding to not more than 0.018% of ionic chloride
`is found.
`
`Chloroform-insoluble phosphorus-
`.
`Ammonium molybdate solution-[NOTE- Prepare fresh on the ~
`of use.] Dissolve 25 g of ammonium molybdate in 300 mL of Watfr
`(Solution A). Cautiously add 75 mL of sulfuric acid to JOO mL f
`water, cool to room temperature, and dilute with water to 200.0 nd.
`(Solution B). Mix Solution A and Solution B to obtain Ammoniun,
`molybdate solution.
`Hydroquinone solution-Dissolve 0.5 g of hydroquinone in 100
`mL of water, and add one drop of concentrated sulfuric acid. [NOTE-.
`When this solution darkens, discard it and prepare fresh. J
`Sodium sulfite solution-Prepare a solution of sodium sulfite in
`water having a concentration of 200 mg per mL. [NOTE-P~ .
`fresh at the time of use.]
`Phosphorus stock solution-Transfer 0.1824 g of mono~
`potassi~m pho~phate, accuratt:IY weighed, to a I 000-mL volu
`·
`flask, dissolve m and dilute with water to volume, and mix.
`.
`Phosphorus illlermediate solution--Transfer 10.0 mL of Phoi.
`phorus stock solution to a I 00-mL volumetric flask, dilute with wara
`to volume, and mix. Prepare this solution fresh on the day ofust.t
`Phosphorus standard solution-Transfer 10.0 mL of Phosp~
`intermediate solution to a I 00-mL volumetric flask, dilute with
`·
`to volume, and mix.
`Test preparation-Transfer I g of lfosfamide, accurately wei
`to a 100-mL volumetric flask, dissolve in 50 mL of water, dilute
`water to volume, and mix. Transfer 10.0 mL of this solution to'.'
`separatory funnel, and add 5 mL of water. Add 15 mL of chlorof,
`shake vigorously for 30 seconds, allow the layers to separate
`drain, and discard the lower chloroform layer. Repeat this extra · ·
`four times, each time with 15 mL of chloroform, discarding
`chloroform layer after each extraction. Transfer the aqueous
`to a conical flask, wash the separatory funnel with two 5-mL po ·
`of water, and collect all the aqueous washings in the same flask.
`3 mL of sulfuric acid, and heat under a hood until white fumes a
`Remove the flask from the heat, and with swirling, add 0.6 mL·
`hydrogen peroxide. Heat until white fumes reappear. If the solu ·
`not colorless, repeat additions of hydrogen peroxide followed
`heating until all color is gone. Cool to room temperature, add 25
`of water, and cautiously add IO mL of ammonium hydro
`solution. Cool to room temperature, add 2 drops of phenolphth
`TS, and then add hydrochloric acid dropwise until all pink color ,
`disappeared. Transfer the contents of the flask to a I 00-mL ft
`dilute with water to volume, and mix.
`,,
`Blank solution-To 3 mL of sulfuric acid in a second conical
`adding 0.6 mL of hydrogen peroxide, proceed as directed for the
`preparation, beginning with "Heat until white fumes reappear."
`Procedure-Transfer 15.0 mL each of the Test preparation,
`Blank solution, and the Phosphorus standard solution to
`separate 25-mL volumetric flasks. Add 2.5 mL of Ammon·
`molybdate solution to each of the flasks, swirl, and allow to stand
`about 30 seconds. To each of the three flasks in order, rapidly add_
`mL each of Hydroquinone solution and Sodium sulfite so
`Dilute the contents ofeach flask with water to volume, mix, and
`the flasks to stand for 30 minutes. Concomitantly determine
`absorbances of the solutions obtained from the Test preparation
`the Phosphorus standard solution in I-cm cells at the wavelen
`maximum absorbance at about 730 nm, with a suitable spec
`tometer, using the solution obtained from the Blank solution as
`blank. Calculate the percentage of chloroform-insoluble phosp
`in the portion of lfosfamide taken by the formula:
`
`lOO(C / W)(Aul As),
`in which C is the concentration, in µg per mL, of phosphorus iJ!.
`Phosphorus standard solution, Wis the weight, in mg, oflfoS
`.
`taken, and Au and As are the absorbances from the solutions o
`from the Test preparation and the Phosphorus standard sol
`respectively: not more than 0.0415% is found.
`Limit of 2-chloroethylamine hydrochloride--
`.
`Standard solution-Dissolve an accurately weighed quan~dty
`chloroethylamine hydrochloride in N,N-dimethylacetam• e, ·
`dilute quantitatively, and stepwise if necessary, with the same 25
`to obtain a solution having a known concentration of about O.O
`per mL.
`Test solution-Transfer about 100 mg of Ifosfamide, ac_d 1
`weighed, to a flask, add 10.0 mL of N,N-dimethylacetam1 e..
`shake until dissolved.
`.
`u·
`Chromatographic system-The gas chromatograph 1s eQ I'
`with a flame-ionization detector and contains a 2-mm >< ,
`
`column packed with 10
`· hydroxide on 80- to I,
`1118llltained at a tempera
`111 a temperature of 1
`1a11perature of about I 4
`1 flow rate of about 25
`Procedure-Separate I
`fest solution and the St1
`,ecord the chrornatogran
`2-chloroethylamine hyd
`cbloroethylamine hydro,
`die formula:
`
`ei which C is the concent
`
`loride in the Sta
`de taken, and ru
`mine peaks obtained fron
`•tively: not more th,
`iide is found.
`r requirements-~
`it meets the req1
`l endotoxins und,
`at Ifosfamide mu1
`tion of injectal
`rial endotoxins 1
`OTE-Ifosfamid
`fresh daily ar
`e Standard p
`ly.J
`hose-Prepare
`itrile (70 : 30). ~
`under Chromat,
`standard solutic
`eighed, to a I(
`issolve. Dilute
`preparation-'
`tely weighed, to
`ndard solution,
`reparation-Tr
`weighed, to a 2:
`ndard solution,
`tographic syste;
`matograph is ei
`30-cm column t
`per minute. C
`the peak resp
`R, between ifosf
`relative standar,
`.0%.
`Separately in
`reparation and ti
`rd the chromat,
`. Calculate the q
`lfosfamide taken I
`
`2:
`is the concentratic
`ard preparation
`the ifosfamide I
`ay preparation
`
`famide for Injecti,
`t and not more th:
`t of C1H1sCliN20,
`
`Slayback Exhibit 1055, Page 43 of 78
`Slayback v. Eye Therapies - IPR2022-00142
`
`

`

`ide
`
`obunolol Hydrochloride Ophthalmic
`tiOD
`
`Levocarnitine
`
`Official Monographs I Levocarnitine
`
`1119
`
`vobunolol Hydrochloride Ophthalmic Solution
`· s not less than 90.0 percent and not more than
`percent of the labeled amount ofC 17H25N03 • HCL
`" · g and storage---Preserve in tight containers.
`eference standards ( 11 )-USP Levobunolol Hydrochloride
`
`ation-The retention time of the major peak in the
`0gram of the Assay preparation corresponds to that of the
`in the chromatogram of the Standard preparation, as
`in the Assay.
`" crobial effectiveness ( 5 I) : meets the requirements.
`'
`(71 )-It meets the requirements when tested as directed for
`ne Filtration under Test for Sterility of the Product to be
`. d.
`I): between 5.5 and 7.5.
`
`'le phase-Dissolve 990 mg of sodium 1-heptanesulfonate in
`of water, add IO mL of glacial acetic acid and 1100 mL of
`I, mix, pass through a suitable filter having a 1-µm or finer
`, and degas. Make adjustments if necessary (see System
`· ity under Chromatography ( 621) ).
`preparation-Dissolve an accurately weighed quant