THE UNITED STATES PHARMACOPEIA

THE NATIONAL FORMULARY

By authority of the United States Pharmacopeial Convention, Inc., meeting at Washington, D. C., March 8-10, 1990. Prepared by the Committee of Revision and published by the Board of Trustees

Official from January 1, 199 5

UNITED STATES PHARMACOPEIAL CONVENTION, INC. 12601 Twinbrook Parkway, Rockville, MD 20852

IPR2016-01033 SHIRE EX2033, p. 1

t/SP 23

Tolerances-Not less than 75% (Q) of the labeled amount of '2H29N0 2 · C 10H80 3S · H20 and not less than 75% (Q) of the llieled amount of C9H80 4 are dissolved in 60 minutes.

Eormity of dosage units (905): meet the requirements for tent Uniformity with respect to propoxyphene napsylate. salicylic acid-

Ferric chloride-urea reagent-Dissolve by swirling, without ile aid of heat, 60 g of urea in a mixture of 8 mL of ferric oloride solution (6 in 10) and 42 mL of 0.05 N hydrochloric pl. Adjust this solution by the addition of 6 N hydrochloric ~ to a pH of 3.2, if necessary.

Standard preparation-Transfer 15.0 mg of USP Salicylic Acid IS, accurately weighed, to a I 00-mL volumetric flask, add chlo­lillorm to volume, and mix. Transfer 10.0 mL of this solution to alOO-mL volumetric flask containing 20 mL of methanol, 4 drops ti.hydrochloric acid, and 20 mL of a I in 10 solution of glacial ~c acid in ether, dilute with chloroform to volume, and mix.

Test preparation-Pack a pledget of glass wool in the base of •25-X 200-mm chromatographic tube. In a beaker, prepare a llixture of 6 g of chromatographic siliceous earth, 2 mL of freshly ~red Ferric chloride-urea reagent, and 40 mL of chloroform. J11115fer the mixture to the chromatographic tube. Rinse the i.ker with 15 mL of chloroform, transfer to the column, and ..,:k tightly. Place a small amount of glass wool at the top of fl&,column. Weigh accurately a quantity of the finely powdered lllllents of Tablets, equivalent to about 50 mg of aspirin, mix ~ 10 mL of chloroform by stirring for 3 minutes, and transfer jdi the aid of IO mL of chloroform to the chromatographic iilumn. Pass 40 mL of chloroform through the column, rinse

~

p of the chromatographic tube with chloroform, and discard uate. Prepare as a receiver a I 00-mL volumetric flask con­g 20 mL of methanol and 4 drops of hydrochloric acid, and any salicylic acid from the column by passing 20 mL of a

i 10 solution of glacial acetic acid in ether that recently has n saturated with water, followed by 30 mL of chloroform. te the eluate with chloroform to volume, and mix.

Procedure-Concomitantly determine the absorbances of the ard preparation and the Test preparation in I-cm cells at avelength of maximum absorbance at about 306 nm, with

"table spectrophotometer, using as the blank a solvent mixture the same composition as that used for the Standard prepa­

l#ion: the absorbance of the Test preparation does not exceed of the Standard preparation (3.0%, calculated on the basis

labeled content of aspirin). y for propoxyphene napsylate-ernal standard solution-Dissolve n-tricosane in chloroform tain a solution containing about 0.6 mg per mL.

tandard preparation-Transfer 50 mg of USP Propoxyphene ylate RS and 163 mg of USP Aspirin RS, all accurately ed, to a 50-mL volumetric flask. Add 10 mL of acetone,

swirl to dissolve the Reference Standards completely. Dilute water to volume, and mix. Jay preparation-Weigh and finely powder not less than 20 ts. Transfer an accurately weighed portion of the powder, lent to about I 00 mg of propoxyphene napsylate, to a I 00-

volumetric flask containing 20 mL of acetone, and sonicate bout I minute. Dilute the milky solution with water to vol­mix, and filter, discarding the first 20 mL of the filtrate. edure-Transfer 5.0-mL aliquots of the Assay preparation

the Standard preparation to separate 60-mL separators. To add 5.0 mL of sodium carbonate solution (I in 5) and 5.0

of Internal standard solution. Shake vigorously for 5 min­and allow the layers to separate. Drain the chloroform layer gh phase-separating paper, suitably supported in a funnel, a screw-capped test tube. Extract with one 5-mL portion of

form, and drain the chloroform layer through phase-sepa­paper. Evaporate the combined chloroform extracts, using m of dry nitrogen, to a final volume of about 2 mL. Inject tely a suitable volume, equivalent to about 6.4 µg of pro­ene, of the chloroform extracts from the Assay prepa­and the Standard preparation into a suitable gas chro­ph equipped with a flame-ionization detector. The column

· lly 120 cm X 3 mm and is packed with 3% phase G3 port SI A. The temperature of the injection port is 200°, umn temperature is 175°, and the carrier gas is nitrogen gat the rate of about 60 mL per minute. Relative retention

Official Monographs / Propranolol 1327

times are 1.0 for the internal standard, and 1.7 for propoxyphene. In a suitable chromatogram, the resolution factor is not less than 1.0 and the relative standard deviation for five replicate injections of the Standard preparation is not more than 2.0. Calculate the quantity, in mg, of C22H29N0 2 · C 1oHs0 3S · H20 in the portion of Tablets taken by the formula:

(565.74/547.72)(1 OOC)(Ru/ Rs),

in which 565.74 and 547.72 are the molecular weights of pro­poxyphene napsylate monohydrate and anhydrous propoxyphene napsylate, respectively, C is the concentration, in mg per mL, of anhydrous propoxyphene napsylate in the Standard preparation, as determined from the concentration of USP Propoxyphene Nap­sylate RS corrected for moisture content by a titrimetric water determination, and Ru and Rs are the peak response ratios ob­tained from the Assay preparation and the Standard preparation, respectively. Assay for aspirin-

Sodium hydroxide reagent-Dissolve I g of polyoxyethylene (23) lauryl ether in about 100 mL of hot water contained in a 1000-mL volumetric flask. Dilute with water to about 600 mL, and dissolve IO g of sodium hydroxide in this solution. Dilute with water to volume, and mix.

Ferric nitrate reagent-Mix 70 mL of nitric acid with about 600 mL of water contained in a 1000-mL volumetric flask. Dis­solve 40 g of ferric nitrate [Fe(N0 3h · 9H 20] in this solution, dilute with water to volume, and mix.

Standard preparation and Assay preparation-Prepare as di­rected in the Assay for propoxyphene napsylate.

Procedure-Into separate 25-mL volumetric flasks pipet 2 mL each of the Standard preparation and the Assay preparation, and 2 mL of dilute acetone (2 in I 0) to provide the blank. Into each flask pipet 5 mL of Sodium hydroxide reagent, mix by gentle swirling, and allow to stand at room temperature for 8 minutes. Dilute with Ferric nitrate reagent to volume, and mix. Concomitantly determine the absorbances of both solutions against the blank in I-cm cells at the wavelength of maximum absorbance at about 530 nm, taking care to allow the solutions to reach an equilibrium temperature in the cell compartment. The color in­tensity is temperature-dependent. Calculate the quantity, in mg, of C9H 80 4 in the portion taken for the Assay preparation by the formula:

I OOC(Au/As),

in which C is the concentration, in mg per mL, of USP Aspirin RS in the Standard preparation, and Au and As are the absorb­ances of the solutions from the Assay preparation and Standard preparation, respectively.

Propranolol Hydrochloride

OH

o~~1

cH3 • HCI

CH3

C16H21N02·HCI 295.81 2-Propanol, 1-[ ( 1-methylethyl)amino ]-3-( 1-naphthalenyloxy )-,

hydrochloride, ( ± )-. ( ± )-1-(Isopropylamino)-3-(1-naphthyloxy)-2-propanol hydro-

chloride [ 318-98-9].

» Propranolol Hydrochloride contains not less than 98.0 percent and not more than 101.5 percent of C 16H21N0 2· HCl, calculated on the dried basis. IPR2016-01033 SHIRE EX2033, p. 2

1328 Propranolol / Official Monographs

Packaging and storage-Preserve in well-closed containers. USP Reference standards ( 11 )-USP Propranolol Hydrochlo­ride RS. Identification-

A: Infrared Absorption (197M). B: The chromatogram of the Assay preparation obtained as

directed in the Assay exhibits a major peak for propranolol, the retention time of which corresponds to that exhibited in the chro­matogram of the Standard preparation obtained as directed in the Assay.

C: It responds to the tests for Chloride (191 ) . Melting range, Class la (741): between 162° and 165°. Specific rotation (781S): between -1.0° and + 1.0°.

Test solution: 40 mg per mL, in water. Loss on drying (731 )-Dry it at 105° for 4 hours: it loses not more than 0.5% of its weight. Residue on ignition ( 281 ) : not more than 0. 1 %.

Organic volatile impurities, Method I (467): meets the require­ments. Assay-

Mobile phase-Dissolve 0.5 g of dodecyl sodium sulfate in 18 mL of 0.15 M phosphoric acid, add 90 mL of acetonitrile and 90 mL of methanol, dilute with water to make 250 mL, mix, and filter through a filter of 0.5 µm or finer porosity. Make adjust­ments if necessary (see System Suitability under Chromatog­raphy (621)).

Standard preparation-Dissolve an accurately weighed quan­tity of USP Propranolol Hydrochloride RS quantitatively in methanol to obtain a stock solution having a known concentration of about I mg per mL. Transfer 5.0 mL of this solution to a 25-mL volumetric flask, dilute with methanol to volume, mix, and filter through a filter of 0.7 µm or finer porosity. This solution contains about 0.2 mg of USP Propranolol Hydrochloride RS per mL.

Resolution solution-Prepare a solution of procainamide hy­drochloride in methanol containing about 0.25 mg per mL. Trans­fer 5 mL of this solution and 5 mL of the stock solution used to prepare the Standard preparation to a 25-mL volumetric flask, dilute with methanol to volume, and mix.

Assay preparation-Transfer about 50 mg of Propranolol Hy­drochloride, accurately weighed, to a 50-mL volumetric flask, add 45 mL of methanol, shake, and sonicate for 5 minutes. Dilute with methanol to volume, mix, and filter through a 0.7-µm or finer porosity filter. Transfer 5.0 mL of this solution to a 25-mL volumetric flask, dilute with methanol to volume, and mix.

Chromatographic system (see Chromatography ( 621) )-The liquid chromatograph is equipped with a 290-nm detector and a 4.6-mm X 25-cm column that contains 5-µm packing L 7. The flow rate is about 1.5 mL per minute. Chromatograph the Res­olution solution, and record the peak responses as directed under Procedure: the relative retention times are about 0.6 for pro­cainamide and 1.0 for propranolol, and the resolution, R, between the procainamide and the propranolol peaks is not less than 2.0. Chromatograph the Standard preparation, and record the peak responses as directed under Procedure: the tailing factor for the propranolol peak is not more than 3.0, 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 chromatograph, record the chromatograms, and measure the re­sponses for the major peaks. Calculate the quantity, in mg, of C 16H 21N0 2 · HCl in the portion of Propranolol Hydrochloride taken by the formula:

250C(ru/ rs),

in which C is the concentration, in mg per mL, of USP Pro­pranolol Hydrochloride RS in the Standard preparation, and ru and rs are the propranolol peak responses obtained from the As­say preparation and the Standard preparation, respectively.

USPI

Propranolol Hydrochloride Extended­release Capsules

» Propranolol Hydrochloride Extended-release~ sules contain not less than 90.0 percent and not d than 110.0 percent of the labeled amount of Cit H21N0 2· HCl. Packaging and storage-Preserve in well-closed containers. Labeling-The labeling states the Drug Release Test with wbii the product complies. USP Reference standards ( 11 )-USP Propranolol Hydrocl,lt. ride RS. Identification-Transfer the contents of a number of Capaulai equivalent to about 160 mg of propranolol hydrochloride, to glass mortar. Add about 5 mL of water, and triturate the mixtlll with a glass pestle. Transfer the suspension to a centrifuge tulle with the aid of about IO mL of water. Add I mL of 1 N sodiua hydroxide, and mix. Add 15 mL of ether, and shake by am; chanical means for about 5 minutes. Centrifuge the mixturc,ui transfer as much of the ether layer as possible to a second• trifuge tube. Add 0.1 mL of hydrochloric acid to the ether• tract, and shake. Centrifuge, and discard the ether layer. A1M 15 mL of ether to the precipitate, and shake by mechanical IJlelll for about 5 minutes. Centrifuge, and discard the ether 1aJmr Dry the precipitate in vacuum at about 45° for 30 minubil Transfer a small amount of the dried precipitate to a mortar, .. grind to a fine powder: the infrared absorption spectru~ mineral oil dispersion of the powder so obtained exhibits · only at the same wavelengths as that of a similar prepara · USP Propranolol Hydrochloride RS. Drug release (724)-

Test I: If the product complies with this test, label the p to indicate that it meets USP Drug Release Test 1.

pH 1.2 buffer solution-Dissolve 2.0 g of sodium chloridf water, add 7 .0 mL of hydrochloric acid, dilute with water liter, and mix.

pH 6.8 buffer solution-Dissolve 21. 72 g of anhydrous dibllle sodium phosphate and 4.94 g of citric acid monohydrate in wul,j dilute with water to I liter, and mix.

Media-Proceed as directed under Method B for De/4"" release (Enteric-coated) Articles, using 900 mL of pH 1.2 biilt solution during the Acid stage, run for 1.5 hours, and use acceptance criteria given under Tolerances. For the Buffer use 900 mL of pH 6.8 buffer solution, run for the times and use the acceptance criteria given under Tolerances.

Apparatus 1: I 00 rpm. Times: 1.5 hours, 4 hours, 8 hours, 14 hours, 24 hours. Procedure-Using filtered portions of the solution under

diluted if necessary, determine the amount of C16H21NOr dissolved, using ultraviolet absorbances at the wavelength of imum absorbance at about 320 nm, with respect to a drawn from 355 nm through 340 nm, by comparison with a dard solution in water having a known concentration of Propranolol Hydrochloride RS.

Tolerances-The percentages of the labeled amount o( H 21N0 2 ·HCI dissolved at the times specified conform ceptance Table 1.

Time (hours) 1.5 4 8

14 24

Amount dissolved not more than 30% between 35% and 60% between 55% and 80% between 70% and 95% between 81 % and 110%

Test 2: If the product complies with this test, label the to indicate that it meets USP Drug Release Test 2.

pH 1.2 buffer solution-Dissolve 2.0 g of sodium chi water, add 7.0 mL of hydrochloric acid, dilute with wa~ liter, and mix.

pH 7.5 buffer solution-Dissolve 6.8 g of monobasic po phosphate and 1.6 g of sodium hydroxide in 900 mL of adjust with 1 N sodium hydroxide to a pH of 7.5, diluto water to I liter, and mix.

IPR2016-01033 SHIRE EX2033, p. 3

Media-Proceed as directed under Method B for Delayed­e (Enteric-coated) Articles-General Drug Release Stan­

' using 900 mL of pH 1.2 buffer solution during the Acid , run for I hour, and use the acceptance criteria given under ranees. For the Buffer stage, use 900 mL of pH 7.5 buffer tion, run for the time specified, and use the acceptance cri­given under Tolerances. paratus 1: 50 rpm.

1mes: I hour, 3 hours, 6 hours, 12 hours. rocedure-Using filtered portions of the solution under test, tcd if necessary, determine the amount of C 16H 21N0 2 -HCI lved, using ultraviolet absorbances at the wavelength of max­

absorbance at about 320 nm, with respect to a baseline wn from 355 nm through 340 nm, by comparison with a Stan­

solution in water having a known concentration of USP ranolol Hydrochloride RS.

olerances-The percentages of the labeled amount of 21 N02 · H Cl dissolved at the times specified conform to Ac­

ance Table 1.

Time (hours) I 3 6

12

Amount dissolved not more than 20% between 20% and 45% between 45% and 80% not less than 80%

ormity of dosage units ( 905): meet the requirements. edure for content uniformity-Transfer the contents of I

ule to a I 00-mL volumetric flask, add about 70 mL of meth-' swirl occasionally for 30 minutes, and sonicate for about I te, and then swirl occasionally for an additional 30 minutes. te with methanol to volume, mix, and centrifuge a portion

the solution. Dilute an accurately measured volume of the solution quantitatively with methanol to obtain a solution ining about 40 µ.g of propranolol hydrochloride per mL. mitantly determine the absorbances of this solution and a rd solution of USP Propranolol Hydrochloride RS in meth-

having a known concentration of about 40 µ.g per mL, in !­cells at the wavelength of maximum absorbance at about 290 with a suitable spectrophotometer, using methanol as the

Calculate the quantity, in mg, of C 16H 21N0 2 • HCI in the ule taken by the formula:

(LC/ D)(Au/ As),

bich Lis the labeled quantity, in mg, of propranolol hydro­'de in the Capsule, C is the concentration, in µ.g per mL, of Propranolol Hydrochloride RS in the Standard solution, D concentration, in µ.g per mL, of the solution from the Cap­based on the labeled quantity per Capsule and the extent ution, and Au and As are the absorbances of the solution the Capsule and the Standard solution, respectively.

y-phate buffer-Dissolve 13.6 g of monobasic potassium

hate in 2 liters of water, and mix. Filter the solution through m or finer porosity filter before use. ile phase-Prepare a suitable degassed mixture of Phos­buffer and acetonitrile (650: 350). Make adjustments if ry (see System Suitability under Chromatography (621) ).

· 111ing solvent-Mix 650 mL of water with 350 mL of acet­. c.

ard preparation-Dissolve an accurately weighed quan­of USP Propranolol Hydrochloride RS in methanol, and di­

ntitatively and stepwise with methanol to obtain a solution a known concentration of about 200 µ.g per mL. Transfer

mL of this solution to a 50.0-mL volumetric flask, add Di-solvent to volume, and mix. ay preparation-Transfer the contents of IO Capsules, ac­y counted, to a 500-mL volumetric flask. Add 300 mL of

I, and swirl by mechanical means for 2 hours. Allow to for about 16 hours, then sonicate for one-half hour, and for one-half hour. Dilute with methanol to volume, mix, ntrifuge a portion of the solution. Dilute an accurately cd volume of the clear solution quantitatively with Dilut­ent to obtain a solution having a concentration of about

of propranolol hydrochloride per mL. atographic system (see Chromatography ( 621) )-The

chromatograph is equipped with a 220-nm detector and a

Official Monographs / Propranolol 1329

4-mm X 15-cm column that contains 5-µ.m packing LI. The flow rate is about 2 mL per minute. Chromatograph the Standard preparation, and record the peak responses as directed under Procedure: the column efficiency determined from the analyte peak is not less than I 000 theoretical plates, the tailing factor for the analyte peak is not more than 3, and the relative standard deviation for replicate injections is not more than 2%.

Procedure-Separately inject equal volumes (about 20 µ.L) of the Standard preparation and the Assay preparation into the chromatograph, record the chromatograms, and measure the re­sponses for the major peaks. The retention time of propranolol is about 5 to 9 minutes. Calculate the quantity, in mg, of Cw H21N0 2 · HCI in each Capsule taken by the formula:

(LC/ D)(ru/rs),

in which Lis the labeled quantity, in mg, of propranolol hydro­chloride in each Capsule, C is the concentration, in µ.g per mL, of USP Propranolol Hydrochloride RS in the Standard prepa­ration, D is the concentration, in µ.g of propranolol hydrochloride per mL, of the Assay preparation, based on the labeled quantity per Capsule, the number of Capsules taken, and the extent of dilution, and ru and rs are the peak responses obtained from the Assay preparation and the Standard preparation, respectively.

Propranolol Hydrochloride Injection

» Propranolol Hydrochloride Injection is a sterile so­lution of Propranolol Hydrochloride in Water for In­jection. It contains not less than 90.0 percent and not more than 110.0 percent of the labeled amount of C16H21N02· HCL Packaging and storage-Preserve in single-dose, light-resistant containers, preferably of Type I glass.

USP Reference standards ( 11 )-USP Propranolol Hydrochlo­ride RS. USP Endotoxin RS. Identification-The chromatogram of the Assay preparation ob­tained as directed in the Assay exhibits a major peak for pro­pranolol, the retention time of which corresponds to that exhibited in the chromatogram of the Standard preparation obtained as directed in the Assay. Bacterial endotoxins (85)-lt contains not more than 55.6 USP Endotoxin Units per mg of propranolol hydrochloride. pH (791): between 2.8 and 4.0.

Other requirements-It meets the requirements under Injections (l ).

Assay-Mobile phase, Standard preparation, Resolution solution, and

Chromatographic system-Prepare as directed in the Assay un­der Propranolol Hydrochloride.

Assay preparation---Transfer an accurately measured volume of Injection, equivalent to about 5 mg of propranolol hydrochlo­ride, to a 25-mL volumetric flask, dilute with methanol to volume, and mix.

Procedure-Proceed as directed for Procedure in the Assay under Propranolol Hydrochloride. Calculate the quantity, in mg, of C 16H 21N0 2 • HCl in each mL of the Injection taken by the formula:

25( C/ V)(ru/rs),

in which C is the concentration, in mg per mL, of USP Pro­pranolol Hydrochloride in the Standard preparation, V is the volume, in mL, of Injection taken, and ru and rs are the pro­pranolol peak responses obtained from the Assay preparation and the Standard preparation, respectively.

IPR2016-01033 SHIRE EX2033, p. 4

1330 Propranolol / Official Monographs

Propranolol Hydrochloride Tablets

» Propranolol Hydrochloride Tablets contain not less than 90.0 percent and not more than 110.0 percent of the labeled amount of C 16H 21N0 2 -HC1. Packaging and storage-Preserve in well-closed, light-resistant containers . USP Reference standards ( 11 )- USP Propranolol Hydrochlo­ride RS . Identification- The chromatogram of the Assay preparation ob­tained as directed in the Assay exhibits a major peak for pro­pranolol, the retention time of which corresponds to that exhibited in the chromatogram of the Standard preparation obtained as directed in the Assay . Dissolution (711 )-

Medium: dilute hydrochloric acid (I in 100); 1000 mL. Apparatus 1: 100 rpm . Time : 30 minutes . Procedure - Determine the amount of C 16H21N0 2 · HCl dis­

solved from ultraviolet absorbances at the wavelength of maxi­mum absorbance at about 289 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 Propranolol Hydrochloride RS in the same medium.

Toleran ces- Not less than 75% (Q) of the labeled amount of C 16H21N0 2 · HCl is dissolved in 30 minutes. Uniformity of dosage units (905) : meet the requirements .

Procedure for content uniformity-Transfer I Tablet to a l 00-mL volumetric flask , add 5 mL of dilute hydrochloric acid (I in 100), and let stand, swirling occasionally , until it is disintegrated . Add about 70 mL of methanol, and sonicate for about l minute. Dilute with methanol to volume, mix, and centrifuge a portion of the solution . Dilute an aliquot of the clear solution quanti­tatively with methanol to provide a solution containing about 40 µg of propranolol hydrochloride per mL. Concomitantly deter­mine the absorbances of this solution and of a solution of USP Propranolol Hydrochloride RS in methanol , at a known concen­tration of about 40 µg per mL, in 1-cm cells at the wavelength of maximum absorbance at about 290 nm, with a suitable spec­trophotometer, using methanol as the blank . Calculate the quan­tity , in mg, of C 16H21N0 2-HC1 in the Tablet by the formula :

(T/D)C(Au/As) ,

in which Tis the labeled quantity, in mg, of propranolol hydro­chloride in the Tablet, D is the concentration, in µg per mL, of the solution from the Tablet, based on the labeled quantity per Tablet and the extent of dilution, C is the concentration, in µg per mL, of USP Propranolol Hydrochloride RS in the Standard solution, and Au and As are the absorbances of the solution from the Tablet and the Standard solution , respectively. Assay-

Mobile phase, Standard preparation, Resolution solution , and Chromatographic system-Prepare as directed in the Assay un­der Propranolol Hydrochloride.

Assay preparation-Weigh and finely powder not less than 20 Tablets. Transfer an accurately weighed portion of the powder, equivalent to about 50 mg of propranolol hydrochloride, to a 50-mL volumetric flask, add 40 mL of methanol, shake, and sonicate for 5 minutes. Dilute with methanol to volume , mix, and filter through a 0. 7-µm or finer porosity filter . Transfer 5.0 mL of this solution to a 25-mL volumetric flask, dilute with methanol to volume, and mix.

Procedure-Proceed as directed for Procedure in the Assay under Propranolol Hydrochloride . Calculate the quantity, in mg, of C 16H21N0 2·HC1 in the portion of Tablets taken by the formula :

250C(ru/ rs),

in which C is the concentration , in mg per mL, of USP Pro­pranolol Hydrochloride in the Standard preparation , and ru and rs are the propranolol peak responses obtained from the Assay preparation and the Standard preparation, respectively.

Propranolol Hydrochloride and Hydrochlorothiazide Extended-release Capsules

» Propranolol Hydrochloride and Hydrochlor azide Extended-release Capsules contain not less 90.0 percent and not more than 110.0 percent of labeled amounts of propranolol hydrochloride ( H 21N0 2 · HCl) and hydrochlorothiazide (C7H8CI 04S2). Packaging and storage-Preserve in well-closed containers. USP Reference standards ( 11 )-USP Propranolo/ Hydr ride RS . USP Hydrochlorothiazide RS . USP 4-Amino-6-cW 1,3-benzenedisu/fonamide RS . Identification-

A: Transfer the contents of a number of Capsules, equi to about 100 mg of hydrochlorothiazide, to a 20-mesh sieve. up any large lumps with the aid of a spatula, and collect powder that passes through the sieve. [NOTE-Retain the terial on the screen for identification test B.] Transfer the that passed through the sieve to a screw-capped, 35-mL cent · tube, add 5 mL of solvent hexane, and shake for 5 min Centrifuge , and discard the solvent. To the residue in the trifuge tube add 10 mL of l N sodium hydroxide, shake, filter, collecting the filtrate in a separator. Wash the filter 5 mL of water, and collect the washing in the separator. Ml 50 mL of ether to the separator, shake for 2 minutes, and allali the phases to separate. Drain the aqueous layer into a bcaail adjust with 6 N hydrochloric acid to a pH of about 2, indtat crystallization by scratching the inner surface of the beaker will a glass rod, and allow to stand until crystallization is com Collect the crystals on a filter, and dry at 105° for 30 min Grind the crystals to a fine powder : the infrared absorption• trum of a mineral oil dispersion of the powder so obtained exbt'Wli maxima only at the same wavelengths as that of a similar aration of USP Hydrochlorothiazide RS .

B: Wash the material retained on the screen from ldt. cation test A with a small amount of water, discarding the ings. Transfer the particles remaining on the screen to a mortar, add about 5 mL of water, and triturate the mixture a glass pestle . Transfer the suspension, with the aid of about mL of water, to a 35-mL screw-capped centrifuge tube, add a l mL of l N sodium hydroxide, and mix. Add about 15 ether, and shake by mechanical means for 5 minutes. Cent· the mixture, and transfer as much of the ether layer as to a second centrifuge tube. Add 0.1 mL of hydrochloric to the ether extract, and shake . Centrifuge, and discard thee Add about 15 mL of ether to the residue, and shake by chanical means for 5 minutes. Centrifuge, and discard the layer. Dry the residue in vacuum at 45° for 30 minutes. the crystals to a fine powder : the infrared absorption s of a mineral oil dispersion of the powder so obtained e maxima only at the same wavelengths as that of a similar aration of USP Propranolol Hydrochloride RS .

C: The chromatogram of the Assay preparation obtai directed under Assay and limit of 4-amino-6-chloro-l, zenedisu/fonamide exhibits major peaks for propranolol h chloride and hydrochlorothiazide, the retention times of correspond to those exhibited in the chromatogram of the Si dard preparation obtained as directed under Assay and Ii 4-amino-6-chloro-1,3-benzenedisu/fonamide. Drug release (724) -

pH 1.5 buffer solution, pH 6.8 buffer solution, Media, Apparatus - Proceed as directed in the test for Drug relttlll der Propranolol Hydrochloride Extended-re/ease Capsule,.

Analytical method-Determine the amounts of hydroc thiazide (C 7H8CIN 30 4S2) and propranolol hydrochloride t H21N0 2·HCI) dissolved, using the following method.

Stock propranolol hydrochloride standard solution­a solution of USP Propranolol Hydrochloride RS in dilu drochloric acid (I in I 00) having a known concentration of 0.4 mg per mL.

IPR2016-01033 SHIRE EX2033, p. 5

Stock hydrochlorothiazide standard solution-Dissolve an ac­,JJ.ately weighed quantity of USP Hydrochlorothiazide RS in :125 N sodium hydroxide to obtain a solution having a concen­Jmtion of about 25 mg per mL. Dilute this solution quantitatively 11th water to obtain a solution having a known concentration of *>ut 0.5 mg per mL.

Standard solutions-Prepare, by combining aliquots of the two k standard solutions and diluting with dilute hydrochloric

ICid (I in 100), solutions bracketing the expected concentration the samples at the various time points. Times: 30 minutes, 0.0625D hours, 0.167 D hours, 0.333D hours,

l583D hours, l .OOD hours. Procedure-Use an automatic analyzer consisting of a liquid

ilunpler, a proportioning pump, two ultraviolet spectrophotome­ln, and a manifold consisting of the components illustrated in lie diagram under Automated Methods of Analysis (16) . Start lcsampler and conduct determinations at a rate of 30 per hour, liing a ratio of about I : 1 for the sample to wash time. Calculate le amounts of C7H 8CIN 30 4S2 and C 16H 21N0 2 -HC1 dissolved '1 comparison with the Standard solutions.

Tolerances (Hydrochlorothiazide)-Use the Acceptance Ta­tit under Dissolution (711). Not less than 80% (Q) of the la­fiefed amount of C7H 8ClN 30 4S2 is dissolved in 30 minutes.

Tolerances (Propranolol Hydrochloride)-The percentages of le labeled amount of C 16H 21N0 2 ·HCI dissolved at the times ipecified conform to Acceptance Table I under Drug Release f124).

Time (hours) 0.0625D 0.167D 0.333D 0.583D I.DOD

Amount dissolved (%) not more than 30% between 35% and 60% between 55% and 80% between 70% and 95% between 83% and 108%

;

ormity of dosage units (905): meet the requirements for ent Uniformity with respect to propranolol hydrochloride to hydrochlorothiazide.

Procedure for content uniformity-Apparatus-Use an automatic analyzer consisting of (I) a 20-

el peristaltic pump; (2) an ultraviolet spectrophotometer 'pped with a 10-mm flow cell and a 293-nm detector; (3) an violet spectrophotometer equipped with a I 0-mm flow cell a 273-nm detector; (4) recording devices for each of the two mentioned detectors; and (5) a manifold consisting of com-nts illustrated in the pertinent diagram in the chapter Au­ted Methods of Analysis (16).

Standard hydrochlorothiazide stock solution-Dissolve an ac­:carately weighed quantity of USP Hydrochlorothiazide RS in

~anol to obtain a solution having a known concentration of lliout 5 mg per mL. ~IStandard propranolol hydrochloride stock solution-Dissolve

:accurately weighed quantity of USP Propranolol Hydrochlo­RS in methanol to obtain a solution having a known con­ation of about 5J mg per mL, J being the ratio of the labeled nt, in mg, of propranolol hydrochloride to the labeled amount,

:mg, of hydrochlorothiazide per Capsule. andard preparation-Transfer 10.0 mL of the Standard hy­hlorothiazide stock solution, 10.0 mL of the Standard pro­lot hydrochloride stock solution, and I 0.0 mL of methanol 100-mL volumetric flask, dilute with 0.12 N hydrochloric to volume, and mix.

tst preparations-Transfer the contents of an appropriate ber of individual Capsules to separate I 00-mL volumetric , rinsing each empty Capsule shell with 2 mL of methanol,

adding the rinsings to the respective volumetric flasks . Add of methanol to each flask, and mix by mechanical means

30 minutes. Dilute the contents of each flask with 0.12 N hloric acid to volume, and mix. edure-With the sampler in the standby position, pump

gents through the system until a stable baseline is achieved. te the sampler, and allow one cycle to pass without intro­the Standard preparation or the Test preparations, then

uce a 5-mL portion of the Standard preparation into the r for the next two cycles and for every sixth cycle there­Disregard the first value for the Standard preparation.

the Test preparations to the sampler at the rate of 30 per , using a ratio of about 1 : 1 for the sample to wash time, to

Official Monographs / Propranolol 1331

follow the second 5-mL portion of the Standard preparation. Record the absorbance values, and calculate each peak value by the difference between peak height and baseline. Calculate the quantity, in mg, of hydrochlorothiazide (C7H8CIN 30 4S2) per Capsule taken by the formula:

1 OOC(Au/ As),

in which C is the concentration, in mg per mL, of USP Hydro­chlorothiazide RS in the Standard preparation, Au is the ab­sorbance at 273 nm of the individual Test preparation, and As is the averaged absorbance at 273 nm of the Standard prepa­rations. Make any necessary correction of the results obtained as follows.

(I) Calculate the correction, F', by the formula:

F' = A/P',

in which A is the weight of active ingredient equivalent to 1 average dosage unit obtained by the Assay procedure, and P' is the weight of active ingredient equivalent to 1 average dosage unit calculated as the mean of the dosage units tested by the Content Uniformity procedure.

(2) If F' is between 0.970 and 1.030, no correction is re­quired.

(3) If F' is not between 0.970 and 1.030, calculate the weight of active ingredient in each dosage unit by multiplying each of the weights found using the special procedure by F '.

Calculate the quantity, in mg, of propranolol hydrochloride (C 16H21N0 2 -HCI) per Capsule by the same formula, in which C is the concentration, in mg per mL, of USP Propranolol Hy­drochloride RS in the Standard preparation, Au is the absorbance at 293 nm of the individual Test preparation, and As is the averaged absorbance at 293 nm of the Standard preparations. Make any necessary correction of the results obtained as directed above. Related compounds-

Tetrabut yl ammonium hydroxide solution, Buffer, Mobile phase, Standard preparation, and Chromatographic system­Prepare as directed in the Assay.

Standard solution-Transfer about 25 mg of USP 4-Arnino-6-chloro-1,3-benzenedisulfonamide RS, accurately weighed, to a 200-mL volumetric flask, add 30 mL of methanol, and swirl to dissolve. Dilute with Buffer to volume, and mix. Dilute an ac­curately measured volume of this solution quantitatively, and stepwise if necessary, with Mobile phase to obtain a solution having a known concentration of about 0.5 µ.g per mL.

Test solution-Use the Assay preparation prepared as di­rected in the Assay .

Procedure-Proceed as directed for Procedure in the Assay, except to inject equal volumes (about 50 µ.L) of the Standard solution and the Test solution. Calculate the percentage of 4-amino-6-chloro-1,3-benzenedisulfonamide in the Capsules taken by the formula:

lOOO(C/ NL)(ru/rs),

in which C is the concentration, in µ.g per mL, of USP 4-Amino-6-chloro-1,3-benzenedisulfonamide RS in the Standard solution, N is the number of Capsules taken to prepare the Test solution, L is the labeled amount, in mg, of hydrochlorothiazide in each Capsule taken, and ru and rs are the peak responses of 4-amino-6-chloro-1,3-benzenedisulfonamide obtained from the Test so­lution and the Standard solution, respectively: not more than 1.0% is present. Assay-

Tetrabutylammonium hydroxide solution-Use a suitable aqueous or methanolic solution having a known concentration of tetrabutylammonium hydroxide.

Buffer-Dissolve 31.25 g of monobasic potassium phosphate in 500 mL of water in a 1000-mL volumetric flask. Add 18.75 mL of phosphoric acid, mix, and add a volume of Tetrabutyl­ammonium hydroxide solution equivalent to about 13 g of tet­rabutylammonium hydroxide. Dilute with water to volume, and mix. Dilute 100 mL of this solution with water to obtain 1000 mL of solution, adjusting, if necessary, with phosphoric acid or ION potassium hydroxide to a pH of 2.4 ± 0.1.

IPR2016-01033 SHIRE EX2033, p. 6

1332 Propranolol / Official Monographs

Mobile phase-Prepare a suitable mixture of Buffer and meth­anol (850: 150). Make adjustments if necessary (see System Suitability under Chromatography (621) ).

Standard hydroch/orothiazide stock solution-Transfer about 25 mg of USP Hydrochlorothiazide RS, accurately weighed, to a 100-mL volumetric flask, add 15 mL of methanol, and sonicate for 5 minutes, adding ice to the bath, if necessary, to maintain the temperature at not more than 20°. Dilute with Buffer to volume, and mix. Use this solution within 3 days.

Standard proprano/ol hydrochloride stock solution-Dissolve an accurately weighed quantity of USP Propranolol Hydrochlo­ride RS in Mobile phase to obtain a solution having a known concentration of about 0.25] mg per mL, J being the ratio of the labeled quantity, in mg, of propranolol hydrochloride to the la­beled quantity, in mg, of hydrochlorothiazide per Capsule.

Standard preparation-Transfer 5.0 mL of Standard hydro­chlorothiazide stock solution and 5.0 mL of Standard propran­olol hydrochloride stock solution to a 25-mL volumetric flask, dilute with Mobile phase to volume, and mix. This solution con­tains about 50 µg of hydrochlorothiazide and 50J µg of propran­olol hydrochloride per mL. Use this solution within 3 days.

Assay preparation-Carefully open an accurately counted number of Capsules, equivalent to about 500 mg of hydrochlo­rothiazide, and transfer the contents and the Capsule shells to a 500-mL volumetric flask. Add 5.0 mL of water to the flask, and allow to stand for 5 minutes. Dilute with methanol to volume, mix, and sonicate for 10 minutes, adding ice to the bath, if nec­essary, to maintain the temperature at not more than 20°. Re­move the flask from the bath, and shake it occasionally for 1 hour. Centrifuge a portion of the contents of the flask, if nec­essary, to obtain a clear solution. Transfer 5.0 mL of the clear solution to a 100-mL volumetric flask, add 10.0 mL of methanol, dilute with Buffer to volume, and mix.

Chromatographic system (see Chromatography (621))-The liquid chromatograph is equipped with a 220-nm detector and a 4-mm X 15-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 under Pro­cedure: the column efficiency determined from the propranolol peak is not less than 2500 theoretical plates when calculated by the formula:

5.545(t I wh12F,

the tailing factor for the hydrochlorothiazide and propranolol peaks is not more than 1.5, and the relative standard deviation for replicate injections is not more than 2.0%.

Procedure-[NOTE-Use peak areas where peak responses are indicated.] Separately inject equal volumes (about 50 µL) of the Standard preparation and the Assay preparation into the chro­matograph, record the chromatograms, and measure the re­sponses for the major peaks. The retention time for propranolol is between 12 and 25 minutes, and the relative retention times are about 0.25 for 4-amino-6-chloro-1,3-benzenedisulfonamide, 0.4 for hydrochlorothiazide, and 1.0 for propranolol. Calculate the quantities, in mg, of hydrochlorothiazide (C7H8CIN 30 4S2) and propranolol hydrochloride (C 16H21N0 2 · HCI) in each Capsule taken by the same formula:

10( C/ N)(ru/ rs),

in which C is the concentration, in µg per mL, of USP Propranolol Hydrochloride RS or USP Hydrochlorothiazide RS in the Stan­dard preparation, N is the number of Capsules taken to prepare the Assay preparation, and ru and rs are the peak responses of the corresponding analyte obtained from the Assay preparation and the Standard preparation, respectively.

Propranolol Hydrochloride and Hydrochlorothiazide Tablets

» Propranolol Hydrochloride and Hydrochlorothi­azide Tablets contain not less than 90.0 percent and

USP1A

not more than 110.0 percent of the labeled amouitl of propranolol hydrochloride (C 16H 21N0 2 · HCI) hydrochlorothiazide ( C7H 8ClN 30 4S2).

Packaging and storage---Preserve in well-closed containers. USP Reference standards ( 11 )-USP Propranolol HydrocW, ride RS. USP Hydrochlorothiazide RS. USP 4-Amino-6-chlfM: 1,3-benzenedisulfonamide RS. Identification-

A: Transfer a quantity of finely powdered Tablets, equi. to about 100 mg of propranolol hydrochloride, to a 50-mL 9 trifuge tube, add 15 mL of water and 1 mL of 1 N sodhiii hydroxide, and mix. Add 20 mL of ether, cap the tube, .. shake by mechanical means for 5 minutes. Centrifuge the m1fr ture, and transfer as much of the ether layer as possible ft.I second centrifuge tube. Add 0.05 mL of hydrochloric ai the ether extract, and shake. Centrifuge, and discard the layer. Add 20 mL of ether to the residue in the tube, and by mechanical means for 5 minutes. Centrifuge, and discard, ether layer. Dry the residue in the tube in vacuum at about for 30 minutes. Transfer a small amount of the dried residlll!ilt a mortar, and grind to fine powder: the infrared absorption8flll' trum of a mineral oil dispersion of the powder exhibits lllllla only at the same wavelengths as that of a similar preparatioaff USP Propranolol Hydrochloride RS.

B: Transfer a quantity of finely powdered Tablets, equivall* to about 100 mg of hydrochlorothiazide, to a 35-mL cen~ tube, add 30 mL of acetone, mix, and allow to stand for minutes, with occasional shaking. Centrifuge, and decant acetone extract into a beaker, discarding the residue in the at trifuge tube. Evaporate the acetone extract on a steam ba~lll dryness, add 10 mL of 0.1 N sodium hydroxide, and mix, 111191 a spatula to dislodge any residue from the beaker. Transferji suspension to a 125-mL separator, and wash the beaker with' 5 mL of water, adding the washing to the separator. Add SO of ether to the separator, shake for 2 minutes, and allow the to separate. Draw off the clear lower layer, filtering it · beaker. Add 1 N hydrochloric acid dropwise with stirring a pH of about 2 is reached. [NOTE-Precipitation occurs d the addition of the acid.] When precipitation is complete, the supernatant liquid, and wash the precipitate with 5 water. Dry the precipitate at 105° for 30 minutes: the inf absorption spectrum of a mineral oil dispersion of the dried cipitate exhibits maxima only at the same wavelengths as of a similar preparation of USP Hydrochlorothiazide RS. Dissolution (711 )-

Medium: 0.1 N hydrochloric acid; 900 mL. Apparatus I: 100 rpm. Time: 30 minutes. . :ff;. Procedure-Filter a portion of the solution under test, l!8:P.

10.0 mL of the filtrate to a suitable capped bottle, add 5. of water, 1.0 mL of 5 N sodium hydroxide, and 25.0 mL heptane. Cap the bottle, shake by mechanical means for S utes, and allow the layers to separate, centrifuging if ne to obtain clear upper (n-heptane) and lower (aqueous) ex Determine the quantity, in µg, of propranolol hydroc (C 16H21N02 · HCI) in each mL of then-heptane extract f ultraviolet absorbance at the wavelength of maximum a at about 293 nm, in comparison with an n-heptane S solution obtained by similarly treating and extracting a of 5.0 mL of an aqueous solution having a known conce of USP Propranolol Hydrochloride RS and 5.0 mL of a 0. sodium hydroxide solution having a known concentration of Hydrochlorothiazide RS, using a blank consisting of the tane extract obtained by similarly treating and extracting mL of water. Determine the quantity, in µg, of hydrochl azide (C7H 8CIN 30 4S2) in each mL of the aqueous extract the ultraviolet absorbance at the wavelength of maxim sorbance at about 273 nm, in comparison with the aqueous remaining from the preparation of the n-heptane Stan lution, using as a blank the aqueous extract remaining f preparation of the n-heptane blank extract. Calculate the tity, in mg, of propranolol hydrochloride dissolved by mul · the quantity, in µg, of propranolol hydrochloride in each the n-heptane extract from the solution under test by 2.25 culate the quantity, in mg, of hydrochlorothiazide dissol

IPR2016-01033 SHIRE EX2033, p. 7

It

,l

f/SP 23

•tiplying the quantity, in µ,g, of hydrochlorothiazide in each IL of the aqueous extract from the solution under test by 1.44.

Tolerances- Not less than 80% (Q) of the labeled amount of CtdI11N0 2 · HCI and not less than 80% (Q) of the labeled amount GfC1HsCIN30 4S2 is dissolved in 30 minutes. Viiformity of dosage units (905): meet the requirements for Content Uniformity with respect to propranolol hydrochloride ~ hydrochlorothiazide.

Procedure for content uniformity-Transfer a Tablet to a suit­.we container, and add 500.0 mL of 0.1 N hydrochloric acid. lake until the Tablet has disintegrated, sonicate for 30 seconds, irate by mechanical means for 30 minutes, and then repeat the l!llication and shaking. Centrifuge a portion of the solution, and lansfer 6.0 mL of the clear supernatant liquid and 15.0 mL of

to a suitable capped bottle. Add 1.0 mL of 5 N sodium xide and 25.0 mL of n-heptane, cap the bottle, shake by anical means for 5 minutes, and allow the layers to separate, fuging, if necessary, to obtain clear upper (n-heptane) and (aqueous) layers (Test solutions). Prepare a similar Stan­

solution by mixing 6.0 mL of 0.1 N hydrochloric acid, 3.0 a of water, 6.0 mL of an aqueous solution having a known ~ntration of USP Propranolol Hydrochloride RS, and 6.0 iiiL of a 0.04 N sodium hydroxide solution having a known con­latration of USP Hydrochlorothiazide RS, and proceeding as kted for the Test solutions, beginning with "Add 1.0 mL of

sodium hydroxide." Prepare similar blank n-heptane and 111cous extracts by adding 6.0 mL of 0.1 N hydrochloric acid tll 15.0 mL of water, and proceeding as directed for the Test lllutions, beginning with "Add 1.0 mL of 5 Nsodium hydroxide."

!mitantly determine the absorbances of the n-heptane Test

on and the n-heptane Standard solution at the wavelength aximum absorbance at about 293 nm, using the n-heptane

extract to set the instrument. Calculate the quantity, in of propranolol hydrochloride (C 16H 21N0 2 · HCI) in the Tab-ken by the formula:

(12.5/6)(C)(Au/ As),

•which C is the concentration, in µ,g per mL, of USP Propranolol IJydrochloride RS in the n-heptane Standard solution, and Au ~ As are the absorbances at 293 nm of the n-heptane Test iiution and the n-heptane Standard solution, respectively. Con-

E'tantly determine the absorbances of the aqueous Test so­

and the aqueous Standard solution at the wavelength of um absorbance at about 273 nm, using the aqueous blank t to set the instrument. Calculate the quantity, in mg, of hlorothiazide (C7H8CIN 30 4S2) in the Tablet taken by the

ll'mula:

(11/6)(C)(Au/ As),

which C is the concentration, in µ,g per mL, of USP Hydro­thiazide RS in the aqueous Standard solution, and Au and

are the absorbances at 273 nm of the aqueous Test solution the aqueous Standard solution, respectively.

and limit of 4-amino-6-cbloro-1,3-benzenedisulfonamide--­tlrabut y/ammonium hydroxide solution-Use a suitable

us or methanolic solution having a known concentration of butylammonium hydroxide. iffer-Dissolve 6.8 g of monobasic potassium phosphate in mL of water in a 2000-mL volumetric flask. Add 3.4 mL osphoric acid and a volume of Tetrabutylammonium hy­ide solution equivalent to about 2.6 g of tetrabutylammon-hydroxide, dilute with water to volume, and mix. Adjust, if

ary, with phosphoric acid or IO N potassium hydroxide to of 2.5 ± 0.1, and filter through a 0.5-µ,m or finer porosity

i/e phase- Prepare a suitable mixture of Buffer and meth­(850: 150). Make adjustments if necessary (see System

ability under Chromatography ( 621)) so that the retention of propranolol is between 12 and 25 minutes. andard hydrochlorothiazide stock solution - Transfer about g of USP Hydrochlorothiazide RS, accurately weighed, to mL volumetric flask, add 15 mL of methanol, and mix to

ve. Dilute with Buffer to volume, and mix. ard proprano/ol hydrochloride stock solution - Dissolve rately weighed quantity of USP Propranolol Hydrochlo­

RS in Mobile phase to obtain a solution having a known

Official Monographs / Propranolol 1333

concentration of about 0.251 mg per mL, J being the ratio of the labeled quantity, in mg, of propranolol hydrochloride to the la­beled quantity, in mg, of hydrochlorothiazide per Tablet.

Standard preparation - Transfer 5.0 mL of Standard hydro­ch/orothiazide stock solution and 5.0 mL of Standard propran­olol hydrochloride stock solution to a 25-mL volumetric flask, dilute with Mobile phase to volume, and mix. This solution con­tains about 50 µ,g of hydrochlorothiazide and 50J µ,g of propran­olol hydrochloride per mL.

Standard 4-amino-6-chloro-1,3-benzenedisulfonamide solu­tion-Dissolve an accurately weighed quantity of USP 4-Amino-6-chloro-1,3-benzenedisulfonamide RS in methanol to obtain a solution having a known concentration of about 0.5 mg per mL. Dilute an accurately measured volume of this solution quanti­tatively, and stepwise if necessary, with Mobile phase to obtain a solution having a known concentration of about 0.5 µ,g per mL.

Assay preparation - Weigh and finely powder not Jess than 20 Tablets. Transfer an accurately weighed portion of the powder, equivalent to about 25 mg of hydrochlorothiazide, to a 500-mL volumetric flask. Add 5 mL of water, mix, and allow to stand for 5 minutes, with occasional swirling. Add 75 mL of methanol, mix, and sonicate for l O minutes, with occasional swirling, adding ice to the bath, if necessary, to maintain the temperature at not more than 20°. Add about 350 mL of Buffer to the flask, and sonicate for 10 minutes, with occasional swirling, maintaining the temperature of the bath at not more than 20° . Dilute with Buffer to volume, and mix. Centrifuge a portion of this solution, if necessary, to obtain a clear solution (Assay preparation) .

Chromatographic system (see Chromatography (621))-The liquid chromatograph is equipped with a 270-nm detector and a 4-mm X 15-cm column that contains 5-µ,m packing Ll. The flow rate is about 1.5 mL per minute. Chromatograph the Standard preparation, and record the peak responses as directed under Procedure: the column efficiency determined from the propran­olol peak is not Jess than 2500 theoretical plates, the tailing factor for the propranolol and hydrochlorothiazide peaks is not more than 1.5, and the relative standard deviation for replicate injec­tions is not more than 2.0%. Chromatograph the Standard 4-amino-6-chloro-J ,3-benzenedisulfonamide solution, and record the peak responses as directed under Procedure: the relative standard deviation for replicate injections is not more than 5.0%.

Procedure - [NOTE-Use peak areas where peak responses are indicated.] Separately inject equal volumes .(about 50 µ,L) of the Standard preparation, the Standard 4-amino-6-chloro-1,3-ben­zenedisulfonamide solution, and the Assay preparation into the chromatograph, record the chromatograms, and measure the re­sponses for the major peaks. The relative retention times are about 0.25 for 4-amino-6-chloro-1,3-benzenedisulfonamide, 0.4 for hydrochlorothiazide, and 1.0 for propranolol. Calculate the quan­tities, in mg, of propranolol hydrochloride (C 16H 21N0 2 · HCI) and hydrochlorothiazide (C7H 8CJN30 4S2) in the portion of Tablets taken by the same formula:

0.5C(ru/ rs),

in which C is the concentration, in µ,g per mL, of the appropriate Reference Standard in the Standard preparation, and ru and rs are the peak responses of the corresponding analyte obtained from the Assay preparation and the Standard preparation, respec­tively. Calculate the percentage of 4-amino-6-chloro-1,3-ben­zenedisulfonamide in the portion of Tablets taken by the formula :

50(C/ L)(ru/rs),

in which C is the concentration, in µ,g per mL, of USP 4-Amino-6-chloro-1,3-benzenedisulfonamide RS in the Standard 4-amino-6-ch/oro-J ,3-benzenedisulf onamide solution, Lis the amount, in mg, of hydrochlorothiazide in the portion of Tablets taken, based on the labeled amount, and ru and rs are the peak responses of 4-amino-6-chloro-1,3-benzenedisulfonamide obtained from the Assay preparation and the Standard 4-amino-6-chloro-1,3-ben­zenedisulfonamide solution, respectively: not more than 1.0% is found .

Propyl Gallate-see Propyl Gallate NF

IPR2016-01033 SHIRE EX2033, p. 8

1334 Propylene / Official Monographs

Propylene Glycol

OH

H3C ~OH

C3Hs02 76.10 1,2-Propanediol. 1,2-Propanediol [57-55-6].

» Propylene Glycol contains not less than 99.5 per­cent of C3Hs02.

Packaging and storage-Preserve in tight containers.

USP Reference standards ( 11 )- USP Propylene Glycol RS. Identification, Infrared Absorption ( l 97F), on undried specimen. Specific gravity (841): between 1.035 and 1.037. Acidity- Add 1 mL of phenolphthalein TS to 50 mL of water, then add 0.1 N sodium hydroxide until the solution remains pink for 30 seconds. Then add 10 mL of Propylene Glycol, accurately measured, and titrate with 0.10 N sodium hydroxide until the original pink color returns and remains for 30 seconds: not more than 0.20 mL of 0.10 N sodium hydroxide is required. Water, Method I (921): not more than 0.2%. Residue on ignition- Heat 50 g in a tared I 00-mL shallow dish until it ignites, and allow it to burn without further application of heat in a place free from drafts. Cool, moisten the residue with 0.5 mL of sulfuric acid, and ignite to constant weight: the weight of the residue does not exceed 3.5 mg. Chloride (221 )- A 1-mL portion shows no more chloride than corresponds to 0.10 mL of 0.020 N hydrochloric acid (0.007%) .

Sulfate (221 )- A 5.0-mL portion shows no more sulfate than corresponds to 0.30 mL of 0.020 N sulfuric acid (0.006%). Arsenic, Method I (211 ): 3 ppm. Heavy metals (231 )-Mix 4.0 mL with water to make 25 mL: the limit is 5 ppm.

Organic volatile impurities, Method IV (467): meets the require­ments. Assay-

Chromatographic system-The gas chromatograph is equipped with a thermal conductivity detector, and contains a 1-m X 4-mm column packed with 5% Gl6 on support S5. The injection port temperature is 240°, the detector temperature is 250°, and the column temperature is programmed at a rate of 5° per minute from 120° to 200° , and helium is used as the carrier gas. The approximate retention time for propylene glycol is 5. 7 minutes, and the approximate retention times for the 3 isomers of dipro­pylene glycol, when present, are 8.2, 9.0, and 10.2 minutes, re­spectively .

Procedure- Inject a suitable volume, typically about 10 µL, of Propylene Glycol into a suitable gas chromatograph, and re­cord the chromatogram. Calculate the percentage of C3H 80 2 in the Propylene Glycol by dividing the area under the propylene glycol peak by the sum of the areas under all of the peaks, ex­cluding those due to air and water, and multiplying by 100.

Propylene Glycol Alginate-see Propylene Glycol Alginate NF

Propylene Glycol Diacetate- see Propylene Glycol Diacetate NF

Propylene Glycol Monostearate-see Propylene Glycol Monostearate NF

USPS

Propylhexedrine

C 10H21N 155.28 Cyclohexaneethanamine, N,a-dimethyl-, ( ± )-. ( ± )-N,a-Dimethylcyclohexaneethylamine [101-40-6].

» Propylhexedrine contains not less than 98.0 paat, cent and not more than 101.0 percent of C10H21N. Packaging and storage-Preserve in tight containers. Identification-

A: To 3 mL of water contained in a small flask add abolt 0.1 mL of it and 0.5 mL of 1 N hydrochloric acid, and agitat the mixture until clear. Add 20 mL of trinitrophenol TS, i the stopper in the flask, shake vigorously for a few minutes, allow to stand for 2 hours. Filter, wash the precipitate with a 20 mL of cold water, and dry in vacuum at 60° for 4 hours: picrate so obtained melts between 108° and 110° (see Melt Range or Temperature (741) ). (Caution-Picrates may plode .)

B: A solution, prepared as directed in Identification test'&, yields a brown precipitate with iodine TS and a white precipitltlf with mercuric-potassium iodide TS. Specific gravity (841): between 0.848 and 0.852. Assay-Tare a glass-stoppered conical flask containing about mL of water, add quickly about 0.5 mL of Propylhexedrine, again weigh. Add to the contents of the flask 30 mL of tralized alcohol, then add methyl red TS, and titrate with 0.1 sulfuric acid VS. Perform a blank determination, and make necessary correction. Each mL of 0.1 N sulfuric acid is eq alent to 15.53 mg of C 10H 21N .

Propylhexedrine Inhalant

» Propylhexedrine Inhalant consists of cylind · rolls of suitable fibrous material impregnated Propylhexedrine, usually aromatized, and cont · in a suitable inhaler. The inhaler contains not than 90.0 percent and not more than 125.0 per of the labeled amount of C 10H 21N. Packaging and storage-Preserve in tight containers (i and avoid exposure to excessive heat. Identification-Place the contents of I inhaler in a gl pered flask, add 50 mL of methanol, and allow to stand hour with frequent agitation. Filter, pressing out the roll filter. Add to the filtrate 1 N hydrochloric acid until it is s · acid to moistened litmus paper, then add 30 mL of water, evaporate to about 20 mL. Cool, transfer to a small se and shake with 10 mL of ether. Withdraw the water layer, it on a steam bath to expel any ether, and dilute to about 25 From 10 mL of the solution, precipitate the propylhexedrine trinitrophenol TS as directed in Identification test A under pylhexedrine: the propylhexedrine picrate so obtained me! tween 108° and 110° (see Melting Range or Temperature (7 (Caution-Picrates may explode.) Assay-Place the contents of 2 inhalers of Inhalant in the t · of a continuous-extraction apparatus, and quickly assem apparatus. Rinse each of the emptied inhalers with about of methanol, pouring the rinsings through the condenser in extraction flask. Add through the condenser 20 mL to 3 of methanol, and extract for 15 to 20 cycles. Cool the e transfer it completely with the aid of small portions of m to a 100-mL volumetric flask, dilute with methanol to v and mix. To 50.0 mL of the solution add 25.0 mL of sulfuric acid VS, and evaporate to about 40 mL. Cool, add

IPR2016-01033 SHIRE EX2033, p. 9