B, chemical structure, molecular formula, Reference Standards

Bacterial Alkaline Protease Preparation— Use a suitable grade.

[NOTE—Asuitable grade is commercially available as “Protex 6L”from Genencor,www.genencor.com,or as “Optimase Enzyme”from Solvay Enzymes Inc.,www.solvaypharmaceuticals.com.]

Barbital Sodium, C8H11N2NaO3—206.2[144-02-5]—White,crystalline powder or colorless crystals.Freely soluble in water;slightly soluble in alcohol.Use a suitable reagent grade.

Barium Acetate, C4H6BaO4—255.43[543-80-6]—Use ACSreagent grade.

Barium Chloride, BaCl2·2H2O—244.26—Use ACSreagent grade.

Barium Chloride,Anhydrous, BaCl2—208.23—This may be made by drying barium chloride in thin layers at 125

until the loss in weight between two successive,3-hour drying periods does not exceed 1%.

Barium Chloride Dihydrate —Use Barium Chloride.

Barium Hydroxide, Ba(OH)2·8H2O—315.46—Use ACSreagent grade.

Barium Nitrate, Ba(NO3)2—261.34—Use ACSreagent grade.

Basic Fuchsin —See Fuchsin,Basic.

Beef Extract —Aconcentrate from beef broth obtained by extraction from fresh,sound,lean beef by means of cooking with water and evaporating the broth at a low temperature,usually in vacuum,until a thick,pasty residue results.Yellowish brown to dark brown,slightly acid,pasty mass having an agreeable meat-like odor.Store it in tight,light-resistant containers.

For the following tests,prepare a test solutionby dissolving 25g in water to make 250mLof a practically clear and practically sediment-free solution.

Assay for nitrogen content of alcohol-soluble substances— Place a portion of the alcohol filtrate and washings remaining from the test for Alcohol-insoluble substances,corresponding to 1g of the alcohol-soluble solids,in a 500-mL Kjeldahl flask.Add about 10g of powdered potassium sulfate and 20mLof sulfuric acid.Heat the mixture at a low temperature until frothing ceases,then raise the temperature and boil until the mixture acquires a pale yellow color or becomes practically colorless.Cool the flask,add about 250mLof water,and cautiously add sodium hydroxide solution (3in 10)until the contents are alkaline,then add 5mLmore.Connect the flask at once by means of a spray trap to a condenser,the lower outlet tube of which dips beneath the surface of 50.0mLof 0.1Nsulfuric acid VScontained in a receiving flask.Distill the mixture until about 100mLof distillate has been collected in the acid.Add methyl red TS,and titrate the excess acid with 0.1Nsodium hydroxide VS.Each mLof 0.1Nsulfuric acid is equivalent to 1.401mg of N.Not less than 60mg of nitrogen is found.

Assay for nitrogen as ammonia— To 100mLof test solution,contained in a 500-mL Kjeldahl flask,add 5g of barium carbonate and 100mLof water,and by means of a spray trap,connect the flask to a condenser,the lower outlet tube of which dips beneath the surface of 50.0mLof 0.1Nsulfuric acid VScontained in a receiving flask.Distill the mixture until about 100mLof distillate has been collected,add methyl red TS,and titrate the excess acid with 0.1Nsodium hydroxide VS.Each mLof 0.1Nsulfuric acid is equivalent to 1.703mg of NH3.The amount of ammonia found does not exceed 0.35%of the total solids in the portion of test solutiontaken.

Total solids— Distribute 10mLof test solutionover clean,dry sand or asbestos,tared in a porcelain dish,and dry at 105

for 16hours:the residue weighs not less than 750mg (75%).

Residue on ignition— Incinerate the residue obtained in the test for Total solidsby heating the dish to a dull-red heat:the residue does not exceed 30%of the total solids.

Chlorides calculated as sodium chloride— Dissolve the ash obtained in the test for Residue on ignitionin about 50mLof water,and carefully transfer to a 100-mLvolumetric flask.Add to the solution a few drops of nitric acid and 10.0mLof 0.1Nsilver nitrate VS.Add water to volume,and mix.Filter into a dry flask through a dry filter,rejecting the first 10mLof the filtrate.To 50.0mLof the subsequent filtrate add 1mLof ferric ammonium sulfate TS,and titrate with 0.1Nammonium thiocyanate VS.Each mLof 0.1Nsilver nitrate is equivalent to 5.844mg of NaCl.The weight of chlorides calculated as sodium chloride obtained,when multiplied by 2,does not exceed 6%of the total solids.

Alcohol-insoluble substances— Transfer 25mLof test solutionto a 100-mLconical flask,add 50mLof alcohol,and shake thoroughly.Collect the precipitate on a counterpoised filter,wash it three times with a mixture of 2volumes of alcohol and 1volume of water,and dry at 105

for 2hours:the weight of the precipitate,representing the alcohol-insoluble solids,does not exceed 10%of the total solids in the portion of test solutiontaken.

Nitrate— Boil 10mLof test solutionfor 1minute with 1.5g of activated charcoal,add water to replace that lost by evaporation,filter,and add 1drop of the filtrate to 3drops of a 1in 100solution of diphenylamine in sulfuric acid:no blue color is produced.

Benzaldehyde, C7H6O—106.12—Colorless,strongly refractive liquid having an odor resembling that of bitter almond oil.Soluble in water;miscible with alcohol,with ether,and with fixed and volatile oils.

Assay— Pipet about 1mLinto a tared,glass-stoppered weighing bottle,and weigh accurately.Loosen the stopper,and transfer both the weighing bottle and its contents to a 250-mLconical flask containing 25mLof a hydro-alcoholic solution of hydroxylamine hydrochloride (prepared by dissolving 34.7g of hydroxylamine hydrochloride in 160mLof water,then adding alcohol to make 1000mL,and neutralizing to bromophenol blue by the addition of sodium hydroxide TS).Using a graduated cylinder to measure the volume,rinse the sides of the flask with an additional 50mLof this reagent solution.Allow the solution to stand for 10minutes,add 1mLof bromophenol blue TS,and titrate the liberated hydrochloric acid with 1Nsodium hydroxide VS.Perform a blank determination with the same quantities of the same reagents,and make any necessary correction.Each mLof 1Nsodium hydroxide consumed is equivalent to 106.1mg of C7H6O.Not less than 98%is found.

Specific gravity á841ñ: between 1.041and 1.046.

Refractive index á831ñ: between 1.5440and 1.5465at 20

Hydrocyanic acid— Shake 0.5mLwith 5mLof water,add 0.5mLof sodium hydroxide TSand 0.1mLof ferrous sulfate TS,and warm the mixture gently.Add a slight excess of hydrochloric acid:no greenish blue color or blue precipitate is observed within 15minutes.

Benzalkonium Chloride —Use Benzalkonium Chloride(NFmonograph).

Add the following:

Benzamidine Hydrochloride Hydrate, C7H8N2·HCl—156.6[1670-14-0]—White to off-white powder.Use a suitable grade.

[NOTE—Asuitable grade is available from Sigma-Aldrich,www.sigma-aldrich.com.]

USP28

Benzanilide, C13H11NO—197.23—Off-white,light gray to grayish green powder.Insoluble in water;sparingly soluble in alcohol;slightly soluble in ether.

Melting range á741ñ: between 162

and 165

Solubility in acetone— A1.0-g portion dissolves completely in 50mLof acetone to yield a clear solution.

Benzene, C6H6—78.11—Use ACSreagent grade.

Benzenesulfonamide, C6H5SO2NH2—157.19—White to pale beige crystals.

Melting range á741ñ: between 150

and 153

Benzenesulfonyl Chloride, C6H5SO2Cl—176.62—Colorless,oily liquid.Insoluble in cold water;soluble in alcohol and in ether.Solidifies at 0

Melting range á741ñ: between 14

and 17

Boiling range(Reagent test): between 251

and 252

Benzhydrol (a-Phenylbenzenemethanol), C13H12O—184.23—White to pale yellow crystals.Very slightly soluble in water;soluble in alcohol,in ether,and in chloroform.

Melting range á741ñ: between 65

and 67

,but the range between beginning and end of melting does not exceed 2

Benzoic Acid, C6H5COOH—122.12—Use ACSreagent grade.

[NOTE—Benzoic Acid of a quality suitable as a primary standard is available from the National Institute of Standards and Technology,Office of Standard Reference Materials,www.nist.gov,as standard sample No.350.]

Benzophenone, (C6H5)2CO—182.22—White,crystalline powder.

Melting range á741ñ: between 47

and 49

p-Benzoquinone, C6H4O2—108.09—Dark yellow powder having a green cast.Slightly soluble in water;soluble in alcohol,in ether,and in fixed alkali solutions.May darken on standing.Darkened material may be purified by sublimation in vacuum.

Melting range á741ñ: between 113

and 115

3-Benzoylbenzoic Acid, C14H10O3—226.23—White to off-white powder.

Assay— Prepare a mixture of 1%trifluoroacetic acid in water and 1%trifluoroacetic acid in acetonitrile (55:45)for the mobile phase.Inject about 20µLinto a suitable liquid chromatograph (see Chromatography á621ñ)equipped with a 230-nm detector and a 4.6-mm ×15-cm column that contains packing L1.The flow rate is about 1.5mLper minute.The area of the C14H10O3peak is not less than 98.5%of the total peak area.

Benzoyl Chloride, C6H5COCl—140.57—Use ACSreagent grade.

N-Benzoyl-L-arginine Ethyl Ester Hydrochloride, C15H22N4O3·HCl—342.82—Determine suitability for use as a substrate as directed under Crystallized Trypsin(USPmonograph).

Benzoylformic Acid (phenylglyoxylic acid) C6H5COCO2H—150.14—Powder.Soluble in methanol.

Melting range á741ñ: between 62

and 67

Benzphetamine Hydrochloride, C17H21N·HCl—275.82—White to off-white,odorless,crystalline powder.Freely soluble in water,in alcohol,and in chloroform;slightly soluble in ether.

Assay— Dissolve about 500mg,accurately weighed,in a mixture of 50mLof glacial acetic acid and 10mLof mercuric acetate TS,add 1drop of crystal violet TS,and titrate with 0.1Nperchloric acid VSto a blue-green endpoint.Perform a blank determination,and make any necessary correction.Each mLof 0.1Nperchloric acid is equivalent to 27.58mg of C17H21N·HCl.Between 98.0%and 101.0%,calculated on the dried basis,is found.

Melting range á741ñ: between 152

and 158

Specific rotation á781ñ: between +22

and +26

,determined in a solution containing 200mg in 10mL,the specimen having been previously dried in vacuum at 60

for 3hours.

Loss on drying á731ñ— Dry in vacuum at 60

for 3hours:it loses not more than 1%of its weight.

Residue on ignition á281ñ: not more than 0.2%.

2-Benzylaminopyridine, C12H12N2—184.24—Use a suitable grade.

1-Benzylimidazole, C10H10N2—158.20—White crystals.

Assay— Transfer about 40mg,accurately weighed,to a 100-mLbeaker.Dissolve in 50mLof glacial acetic acid.Titrate with 0.1Nperchloric acid VS,determining the endpoint potentiometrically using a combination calomel-platinum electrode.Perform a blank determination and make any necessary correction.Each mLof 0.1Nperchloric acid is equivalent to 15.82mg of C10H10N2.Not less than 99%is found.

Benzyltrimethylammonium Chloride, C6H5CH2N(CH3)3Cl—185.69—Available as a 60percent aqueous solution.Is clear and is colorless or not more than slightly yellow,and has a slight amine-like odor.

Assay— Pipet 2mLinto a 50-mLvolumetric flask,and add water to volume.Pipet 20mLof the solution into a 125-mLconical flask,add about 30mLof water,then add 0.25mLof dichlorofluorescein TS,and titrate with 0.1Nsilver nitrate VS.Each mLof 0.1Nsilver nitrate is equivalent to 18.57mg of C6H5CH2N(CH3)3Cl.Between 59.5%and 60.5%is found.

Beta-lactamase —Beta-lactamase is an enzyme produced by a variety of bacteria,but is usually obtained from culture filtrates of a strain of Bacillus cereus.It has the specific property of inactivating penicillins and cephalosporins by splitting the bond linking the nitrogen of the thiazolidine to the adjacent carbonyl carbon.

It occurs in the form of small,brown,easily pulverizable pieces or granules.Freely soluble in water,forming a slightly opalescent solution that is practically neutral to litmus paper.Is precipitated from its water solutions by acetone,by alcohol,and by dioxane,and is inactivated by contact with these solvents.Is rapidly inactivated by ethyl acetate and is irreversibly destroyed at a temperature of about 80

Beta-lactamase is assayed by a procedure depending upon a determination of the amount of penicillin Gpotassium or penicillin Gsodium destroyed at a pHof 7.0in a solution of such concentration that the inactivation proceeds as a zero-order reaction.

Betanaphthol —See 2-Naphthol.

Bibenzyl (Dibenzyl), C14H14—182.26—Colorless crystals.Freely soluble in chloroform and in ether;sparingly soluble in alcohol;practically insoluble in water.

Melting range á741ñ: between 53

and 55

Bile Salts —Aconcentrate of beef bile,the principal constituent of which is sodium desoxycholate,determined as cholic acid.Soluble in water and in alcohol;the solutions foam strongly when shaken.

Insoluble substances— Dissolve 5g in 100mLof dilute alcohol (84in 100),warming if necessary to aid solution.Filter within 15minutes through a tared filter,and wash with small portions of the dilute alcohol until the last washing is colorless or practically so,then dry the residue at 105

for 1hour,and weigh:the weight of the residue does not exceed 0.1%.

Assay—

STANDARD CHOLIC ACID SOLUTION— Dissolve 50.0mg of cholic acid,accurately weighed,in dilute acetic acid (6in 10)to make 100mL,and mix.Store in a refrigerator.

PROCEDURE— Dissolve 1.0g,accurately weighed,in 50mLof dilute acetic acid (6in 10).Filter the solution,if necessary,into a 100-mLvolumetric flask,wash the original container and the filter with small portions of dilute acetic acid (6in 10),add the same acetic acid to volume,and mix.Dilute 10mLof this solution,accurately measured,with dilute acetic acid (6in 10)to make 100mL,and mix.

Pipet 1mLeach of the Standard Cholic Acid Solution and the solution of the Bile Salts into two matched test tubes.To each tube add 1mL,accurately measured,of freshly prepared furfural solution (1in 100),immediately place the tubes in an ice-bath for 5minutes,then add to each tube 13mL,accurately measured,of dilute sulfuric acid,made by cautiously mixing 50mLof sulfuric acid with 65mLof water.Mix the contents of the tubes,and place them in a water bath maintained at a temperature of 70

for 10minutes.Immediately transfer the tubes to an ice-bath for 2minutes,then determine the absorbance of each solution at the wavelength of maximum absorbance at about 670nm,with a suitable spectrophotometer.Calculate the quantity,in mg,of cholic acid (C24H40O5)in the weight of the Bile Salts taken by the formula:

500(AU/AS),

in which AUand ASare the absorbances of the solutions from the Bile Salts and the Standard Cholic Acid Solution,respectively.Not less than 45%of cholic acid is found.

Biphenyl, C12H10—154.21—Colorless to white crystals or crystalline powder,having a pleasant odor.Insoluble in water;soluble in alcohol and in ether.Boils at about 254

Melting range á741ñ: between 68

and 72

2,2¢-Bipyridine (a,a¢-Dipyridyl), C10H8N2—156.18—White or pink,crystalline powder.Soluble in water and in alcohol.Melts at about 69

,and boils at about 272

Sensitiveness— Prepare the following solutions:(A)—Dissolve 350mg of ferrous ammonium sulfate in 50mLof water containing 1mLof sulfuric acid,and add 500mg of hydrazine sulfate,then add water to make 500mL.For use,dilute this solution with water in the ratio of 1in 100mL.(B)—Dissolve 8.3g of sodium acetate and 12mLof glacial acetic acid in water to make 100mL.Add 1mLof a solution of the specimen (1in 1000)to a mixture of 10mLof water and 1mLof each of solutions Aand B:a pink color results immediately.

Solubility— A100-mg portion dissolves completely in 10mLof water.

Residue on ignition(Reagent test): not more than 0.2%.

4,4¢-Bis(4-amino-1-naphthylazo)-2,2¢-stilbenedisulfonic Acid, C34H26N6O6S2—678.74—Use a suitable grade.

[NOTE—Asuitable grade is available from TCI America,www.tciamerica.com.]

Bis(2-ethylhexyl)Maleate, C20H36O4—340.50—Colorless to pale yellow,clear liquid.Miscible with acetone and with alcohol.Specific gravity about 0.945.

Assay— Place about 2.5g,accurately weighed,in a 250-mLflask,add 50.0mLof 0.5Nalcoholic potassium hydroxide VS,and reflux for 45minutes.Cool,add 0.5mLof phenolphthalein TS,and titrate the excess alkali with 0.5Nhydrochloric acid VS.Perform a blank determination at the same time,using the same amount of 0.5Nalcoholic potassium hydroxide (See Residual Titrationsunder Titrimetry á541ñ).The difference,in mL,between the volumes of 0.5Nhydrochloric acid consumed in the test titration and blank titration,multiplied by 85.1,represents the quantity,in mg,of bis(2-ethylhexyl)maleate in the portion taken.Not less than 97%is found.

Bis(2-ethylhexyl)Phthalate, C6H4-1,2-[COOCH2(C2H5)CH(CH2)3CH3]2—390.56—Colorless to light yellow liquid.

Refractive index á831ñ: between 1.4855and 1.4875,at 20

Bis(2-ethylhexyl)Sebacate (Dioctyl Sebacate), C8H17OOC(CH2)8COOC8H17—426.67—Pale straw-colored liquid.Insoluble in water.Refractive index about 1.448.Suitable for use in gas chromatography.

Specific gravity,20

/20

á841ñ: between 0.913and 0.917.

Boiling range: between 243

and 248

at 5mm of mercury.

[NOTE—Asuitable grade is “Dioctyl Sebacate,”available from Sigma-Aldrich,www.sigma-aldrich.com.]

Bis(2-ethylhexyl)(phosphoric Acid) [Bis-(2-ethylhexyl)Phosphate], [CH3(CH2)3CH(C2H5)CH2]2HPO4—322.42—Light yellow,viscous liquid.Insoluble in water;freely soluble in chloroform and in ethyl acetate.Refractive index:about 1.443.Specific gravity:about 0.997.

Assay— Dissolve about 250mg,accurately weighed,in 50mLof dimethylformamide,add 3drops of a 1in 100solution of thymol blue in dimethylformamide,and titrate with 0.1Nsodium methoxide VSto a blue endpoint.Perform a blank determination,and make any necessary correction.Each mLof 0.1Nsodium methoxide is equivalent to 32.24mg of (C8H17)2HPO4.Between 95%and 105%is found.

Solubility— One volume dissolves in 9volumes of chloroform to yield a clear solution,and 1volume dissolves in 9volumes of ethyl acetate to yield a clear solution.

Color— A1in 100solution in chloroform exhibits an absorptivity of not more than 0.03at 420nm.

Bismuth Nitrate Pentahydrate, Bi(NO3)3·5H2O—485.07[10035-06-0]—Use ACSreagent grade.

Bismuth Sulfite Agar —Use a suitable grade.

Bis(trimethylsilyl)acetamide (N,O-Bis(trimethylsilyl)-acetamide;BSA), CH3CON[Si(CH3)3]2—203.43—Clear,colorless liquid.Readily hydrolyzes when exposed to moist air.Handle under nitrogen,and store in a cool place.

Assay— Not less than 90%of CH3CON[Si(CH3)3]2,a suitable gas chromatograph equipped with a thermal conductivity detector being used.The following conditions are suitable and provide a retention time of approximately 15minutes.

COLUMN: 3-mm ×1.83-m stainless steel containing 5%phase G1on support S1A.

INJECTION TEMPERATURE: 160

COLUMN TEMPERATURE: 90

,programmed to rise 4

per minute to 160

CARRIER GAS: Helium.

Refractive index á831ñ: between 1.4150and 1.4170at 20

Bis(trimethylsilyl)trifluoroacetamide (BSTFA), CF3CON[Si(CH3)3]2—257.40—Clear,colorless liquid.Readily hydrolyzes when exposed to moist air.Store in a cool place.

Assay— Not less than 98%of CF3CON[Si(CH3)3]2,a suitable gas chromatograph equipped with a thermal conductivity detector being used.The following conditions are suitable and provide a retention time of approximately 15minutes.

COLUMN: 3-mm ×1.83-m stainless steel containing 5%phase G1on support S1A.

INJECTION TEMPERATURE: 170

COLUMN TEMPERATURE: 70

,programmed to rise 4

per minute to 140

CARRIER GAS: Helium.

Refractive index á831ñ: between 1.3820and 1.3860at 20

Bis(trimethylsilyl)trifluoroacetamide with Trimethylchlorosilane —Use a suitable grade.

[NOTE—Asuitable grade is available from Sigma-Aldrich,www.sigma-aldrich.com.]

Blood (for carbon monoxide test in gases)—Use oxalatedor defibrinated bloodof dogs,sheep,cattle,or human beings within 24hours after bleeding.Prepare oxalated bloodby adding 10mg of sodium oxalate to each mLof the freshly drawn blood.

Add the following:

Blood Group A1Red Blood Cells and Blood Group B Red Blood Cells— These cells must be obtained from manufacturers or suppliers licensed by the Center for Biologics Evaluation and Research/Food and Drug Administration.The use of reagents from an unlicensed manufacturer or supplier may invalidate the results.Generally,they are available as part of a kit for ABO Blood Group testing.The cells licensed for use in test tubes can also be used in the microtiter plate method described in the monographs of Red Blood Cellsand Whole Blood.

[NOTE—There are many manufacturers and suppliers of these reagents that are licensed by the Center for Biologics Evaluation and Research Food and Drug Administration.Some examples of licensed manufacturers or suppliers are the following:Gamma Biologics,Houston,TX;and Ortho Diagnostics,Raritan,NJ.]

USP28

Add the following:

Anti-A Blood Grouping Reagent,Anti-B Blood Grouping Reagent,and Anti-AB Blood Grouping Reagent— The reagents can be monoclonal or polyclonal and must be obtained from manufacturers or suppliers licensed by the Center for Biologics Evaluation and Research,Food and Drug Administration for use in microplate tests.The use of reagents from an unlicensed manufacturer or supplier may invalidate the results.Generally,all three reagents are available as part of a kit.

[NOTE—There are many manufacturers and suppliers of these reagents that are licensed by the Center for Biologics Evaluation and Research,Food and Drug Administration.Some examples of licensed manufacturers or suppliers are the following:Gamma Biologics,Houston,TX;and Ortho Diagnostics,Raritan,NJ.]

USP28

Blue Tetrazolium (3,3¢-(3,3¢-Dimethoxy[1,1¢-biphenyl]-4,4¢-diyl)bis[2,5-diphenyl-2H-tetrazolium]dichloride), C40H32Cl2N8O2—727.64—Lemon-yellow crystals.Slightly soluble in water;freely soluble in chloroform and in methanol;insoluble in acetone and in ether.

Solubility in methanol— Dissolve 1g in 100mLof methanol:complete solution results,and the solution is clear.

Color— Transfer a portion of the methanol solution obtained in the preceding test to a 1-cm cell,and determine its absorbance at 525nm,against water as the blank:the absorbance does not exceed 0.20.

Molar absorptivity á851ñ— Its molar absorptivity in methanol,at 252nm,is not less than 50,000.

Suitability test—

STANDARD PREPARATION— Dissolve in alcohol a suitable quantity of USP Hydrocortisone RS,previously dried at 105

for 3hours and accurately weighed,and prepare by stepwise dilution a solution containing about 10µg per mL.

PROCEDURE— Pipet 10-,15-,and 20-mLportions of Standard Preparationinto separate,glass-stoppered,50-mLconical flasks.Add 10mLand 5mL,respectively,of alcohol to the flasks containing the 10-and 15-mLportions of Standard Preparation,and swirl to mix.To each of the flasks,and to a fourth flask containing 20mLof alcohol,add 2.0mLof a solution prepared by dissolving 50mg of blue tetrazolium in 10mLof alcohol,mix,and then add 2.0mLof a solution prepared by diluting 1mLof tetramethylammonium hydroxide TSwith alcohol to 10mL.Mix,allow the flasks to stand in the dark for 90minutes,and determine the absorbances of the three solutions of the steroid standard at 525nm,with a suitable spectrophotometer,using the solution in the fourth flask as the blank.Plot the absorbances on the abscissa and the amount of hydrocortisone on the ordinate scale of arithmetic coordinate paper,and draw the curve of best fit:the absorbance of each solution is proportional to the concentration,and the absorbance of the solution containing 200µg of hydrocortisone is not less than 0.50.

Boric Acid, H3BO3—61.83—Use ACSreagent grade.

(–)-Bornyl Acetate (1,7,7-Trimethylbicyclo[2,2,1]-heptan-2-ol acetate), C12H20O2—196.29[5655-61-8]—Use a suitable grade.

Boron Trifluoride, BF3—67.81—Use a suitable grade.

14%Boron Trifluoride–Methanol —Use a suitable grade.

Add the following:

Bovine Collagen— Use a suitable grade that contains less than 1µg glycosaminoglycan per mg.

[NOTE—Asuitable grade is available from Sigma-Aldrich,www.sigma-aldrich.com.]

USP28

7Percent Bovine Serum Albumin Certified Standard —Available from the National Institute of Standards and Technology,www.nist.gov,as SRM927.

Brilliant Green (Malachite Green G), C27H34N2O4S—482.64—Glistening,golden-yellow crystals.Soluble in water and in alcohol.Absorption maximum:623nm.

Bromelain— Aproteolytic enzyme isolated from pineapple.Use a suitable grade.

Bromine, Br—At.Wt.79.904—Use ACSreagent grade.

a-Bromo-2¢-acetonaphthone (Bromomethyl 2-naphthyl ketone), C12H9BrO—249.10—Tannish–pink crystals.

Melting range á741ñ: between 81

and 83

p-Bromoaniline, C6H6BrN—172.02—White to off-white crystals.Insoluble in water;soluble in alcohol and in ether.

Assay— Transfer about 650mg,accurately weighed,to a suitable container,and dissolve in 50mLof glacial acetic acid TS.Add crystal violet TS,and titrate with 0.1Nperchloric acid VS.Perform a blank determination,and make any necessary correction.Each mLof 0.1Nperchloric acid is equivalent to 17.20mg of C6H6BrN.Not less than 98%is found.

Melting range á741ñ: between 60

and 65

,within a 2

range.

N-Bromosuccinimide, C4H4BrNO2—177.98—White to off-white crystals or powder,having a faint odor.Freely soluble in water,acetone,and glacial acetic acid.[Caution—Highly irritating to eyes,skin,and mucous membranes. ]

Assay— Transfer 200mg,accurately weighed,to a conical flask,add 25mLof 0.5Nalcoholic potassium hydroxide,cover with a watch glass,heat to boiling,and boil for 5minutes.Cool,transfer the solution to a beaker,rinsing the flask with water until the total volume of solution plus rinsings is about 100mL,and add 10mLof glacial acetic acid.Insert suitable electrodes,and titrate with 0.1Nsilver nitrate VS,determining the endpoint potentiometrically.Each mLof 0.1Nsilver nitrate is equivalent to 17.80mg of C4H4BrNO2.Not less than 98%is found.

Brucine Sulfate, (C23H26N2O4)2·H2SO4·7H2O—1013.11—Use ACSreagent grade.

Buffers —See Buffer Solutionsunder Solutions.

1-Butaneboronic Acid, C4H11BO2—101.94—White flakes.

Assay— Dissolve about 50mg,accurately weighed,in 2mLof bis(trimethylsilyl)acetamide,and mix while heating to boiling.Allow the solution to cool,and inject an appropriate specimen into a suitable gas chromatograph (see Chromatography á621ñ)equipped with a flame-ionization detector,helium being used as the carrier gas.The following conditions have been found suitable:a 0.25-mm ×30-m capillary column coated with a 1-µm layer of phase G2;the injection port temperature is maintained at 250

;the detector temperature is maintained at 300

;the column temperature is maintained at 100

and programmed to rise 10

per minute to 250

.The area of the C4H11BO2peak is not less than 99%of the total peak area.

Solubility— Asolution of 50mg in 1mLof alcohol is clear and colorless.

Melting range á741ñ: between 92

and 95

1,3-Butanediol (1,3-Butylene Glycol), C4H10O2—90.12—Viscous,colorless liquid.Very hygroscopic.Soluble in water,in alcohol,in acetone,and in methyl ethyl ketone;practically insoluble in aliphatic hydrocarbons,in benzene,and in toluene.

Assay— Inject an appropriate specimen into a suitable gas chromatograph (see Chromatography á621ñ)equipped with a flame-ionization detector,helium being used as the carrier gas.The following conditions have been found suitable:a 3-mm ×1.8-m stainless steel column containing 20%phase G16on support S1A;the injection port temperature is maintained at 265

;the column temperature is maintained at 150

and programmed to rise 8

per minute to 210

.The area of the butanediol peak is not less than 98%of the total peak area.

Refractive index á831ñ: between 1.4390and 1.4410at 20

2,3-Butanedione (Diacetyl), CH3COCOCH3—86.09—Bright yellow to yellowish–green liquid having a strong,pungent odor.Soluble in water.Miscible with alcohol and with ether.Boils at about 88

.Specific gravity:about 0.98.

Assay—

HYDROXYLAMINE HYDROCHLORIDE SOLUTION— Dissolve 20g of hydroxylamine hydrochloride in 40mLof water,and dilute with alcohol to 400mL.Add,with stirring,300mLof 0.5Nalcoholic potassium hydroxide,and filter.Discard after 2days.

PROCEDURE— Transfer about 1g,accurately weighed,to a glass-stoppered,250-mLflask,add 75.0mLof Hydroxylamine hydrochloride solution,and insert the stopper in the flask.Reflux the mixture for 1hour,then cool to room temperature.Add bromophenol blue TS,and titrate with 0.5Nhydrochloric acid VSto a greenish–yellow endpoint.[NOTE—Alternatively,the solution may be titrated potentiometrically to a pHof 3.4.]Perform a blank test with the same quantities of reagent used for the test specimen,and make any necessary correction.Each mLof 0.5Nhydrochloric acid is equivalent to 43.05mg of CH3COCOCH3.Not less than 97%of CH3COCOCH3is found.

Refractive index á831ñ: between 1.3935and 1.3965,at 20

Congealing temperature á651ñ: between -2.0

and -5.5

Butanol —See Butyl Alcohol.

Butyl Acetate,Normal, CH3COO(CH2)3CH3—116.16—Use ACSreagent grade.

Butyl Alcohol (1-Butanol;Normal Butyl Alcohol), CH3(CH2)2CH2OH—74.12—Use ACSreagent grade.

Butyl Alcohol,Normal —See Butyl Alcohol.

Butyl Alcohol,Secondary (2-Butanol), CH3CH2CH(OH)CH3—74.12—Use ACSreagent grade Isobutyl Alcohol.

Butyl Alcohol,Tertiary, (CH3)3COH—74.12—Use ACSreagent grade tert-butyl alcohol.

4-(Butylamino)benzoic Acid, C11H5NO2—193.25[4740-24-3]—Use a suitable grade.

[NOTE—Asuitable grade is available from Sigma-Aldrich,Inc.,P.O.Box 2060,Milwaukee,WI53201;www.sigma-aldrich.com.]

Butyl Benzoate, C11H14O2—178.23—Thick,oily,colorless to pale yellow liquid.Practically insoluble in water;soluble in alcohol and in ether.

Assay— When examined by gas-liquid chromatography,it shows a purity of not less than 98%.The following conditions have been found suitable for assaying it:use a 3-mm ×1.8-m stainless steel column packed with liquid phase G4on support S1A.Helium is the carrier gas,the injection port temperature is maintained at 180

,the column temperature is 190

,and the flame-ionization detector is maintained at 280

.The retention time is about 15minutes.

Refractive index á831ñ: between 1.4980and 1.5000,at 20

Butyl Ether (n-Dibutyl Ether), C8H18O—130.23—Use a suitable grade.

n-Butyl Chloride (1-Chlorobutane), C4H9Cl—92.57—Clear,colorless,volatile liquid,having a slight,characteristic odor.Highly flammable.Practically insoluble in water.Miscible with alcohol and with ether.

Assay— When examined by gas-liquid chromatography,it shows a purity of not less than 98%.The following conditions have been found suitable for assaying the article:A3-mm ×1.8-m stainless steel column packed with phase G16on support S1.Helium,flowing at a rate of about 40mLper minute,is the carrier gas,the detector temperature is about 310

,the injection port temperature is about 230

,and the column temperature is programmed to rise at 10

per minute from 35

to 150

.Aflame-ionization detector is employed.

Boiling range á721ñ: between 76

and 80

,within a 2

range.

Refractive index á831ñ: between 1.4015and 1.4035at 20

Acidity— Add phenolphthalein TSto 75mL,and titrate with 0.1Npotassium hydroxide in methanol to a faint pink color that persists,with shaking,for 1.5seconds:not more than 0.91mLis required (about 0.005%as HCl).

Water á921ñ: not more than 0.02%,determined by the Titrimetric Method.

Residue after evaporation— Evaporate about 60mL(50g),accurately weighed,in a tared platinum dish on a steam bath,and dry at 105

for 1hour:not more than 0.005%is found.

tert-Butyl Methyl Ether, C5H12O—88.15—Colorless liquid.

Assay— Inject an appropriate specimen into a gas chromatograph (see Chromatography á621ñ)equipped with a flame-ionization detector,helium being used as the carrier gas.The following conditions have been found suitable:a 0.25-mm ×30-m capillary column coated with a 1-µm layer of phase G2;the injection port temperature is maintained at 100

;the detector temperature is maintained at 300

;and the column temperature is maintained at ambient temperature and programmed to rise 10

per minute to 150

.The area of the C5H12Opeak is not less than 99.8%of the total peak area.

Refractive index á831ñ: between 1.367and 1.371at 20

n-Butylamine, CH3CH2CH2CH2NH2—73.14—Colorless to pale yellow,flammable liquid.Miscible with water,with alcohol,and with ether.Store it in tight containers.Specific gravity:about 0.740.

Distilling range,Method Iá721ñ— Not less than 95%distils between 76

and 78

Water,Method Iá921ñ: not more than 1.0%,determined by the Titrimetric Method.

Chloride(Reagent test)— One g (1.5mL)shows not more than 0.01mg of Cl (0.001%).

Acidic impurities— To 50mLadd 5drops of a saturated solution of azo violet in benzene,and titrate quickly with 0.1Nsodium methoxide VSto a deep blue endpoint,observing precautions to prevent absorption of atmospheric carbon dioxide as by use of an atmosphere of nitrogen:not more than 1.0mLof 0.1Nsodium methoxide is required for neutralization.

tert-Butylamine, C3H9CNH2—73.14—Liquid.

Assay— Inject an appropriate volume into a gas chromatograph (see Chromatography á621ñ)equipped with a flame-ionization detector,helium being used as the carrier gas.The following conditions have been found suitable:a 0.25-mm ×30-m capillary column coated with a 1-µm layer of phase G2;the injection port temperature is maintained at 230

;the detector temperature is maintained at 300

;the column temperature is maintained at 130

and programmed to rise 10

per minute to 280

.The area of the C3H9CNH2peak is not less than 99.5%of the total peak area.

Refractive index á831ñ: between 1.3770and 1.3790at 20

n-Butylboronic Acid, C4H9B(OH)2—101.94—Use a suitable grade.

[NOTE—This reagent is usually shipped and stored under water.Before use,remove any excess water by light vacuum filtration.Asuitable grade is available from Sigma-Aldrich,www.sigma-aldrich.com.]

tert-Butyldimethylchlorosilane inN-Methyl-N-tert-butyldimethylsilyltrifluoroacetamide,(1in 100) —Use a suitable grade.

[NOTE—Asuitable grade is available as 99%MTBSTFA,1%TBDMCSfrom Regis Chemical Company,8210Austin Ave.,P.O.Box 519,Morton Grove,IL60053.]

4-tert-Butylphenol, C10H14O—150.22—White,crystalline flakes or needles.Practically insoluble in water;soluble in alcohol and in ether.

Melting range á741ñ: between 98

and 101

Butyric Acid, C4H8O2—88.11—Clear,colorless to faint yellow liquid.Miscible with water and with methanol.

Assay— Weigh accurately about 500mg,transfer to a suitable container,add 30mLof water,and mix.Add 40mLof water,and mix.Add phenolphthalein TS,and titrate with 0.1Nsodium hydroxide VS.Each mLof 0.1Nsodium hydroxide is equivalent to 8.81mg of C4H8O2:not less than 99.0%of C4H8O2is found.

Refractive index á831ñ: about 1.398at 20

Butyrolactone, (Dihydro-2-(3H)-furanone,g-butyrolactone) —86.1—Clear,colorless to practically colorless,oily liquid.Miscible with water.Soluble in methanol and in ether.

Boiling range á721ñ: between 193

and 208

Refractive index á831ñ: about 1.435,at 20

Specific gravity á841ñ: between 1.128and 1.135.