Oil-and Water-Soluble Vitamins with Minerals Capsules
»Oil-and Water-Soluble Vitamins with Minerals Capsules contain one or more of the following oil-soluble vitamins:Vitamin A,Vitamin Das Ergocalciferol (Vitamin D2)or Cholecalciferol (Vitamin D3),Vitamin E,Phytonadione (Vitamin K1),and Beta Carotene;one or more of the following water-soluble vitamins:Ascorbic Acid or its equivalent as Calcium Ascorbate or Sodium Ascorbate,Biotin,Cyanocobalamin,Folic Acid,Niacin or Niacinamide,Dexpanthenol or Panthenol,Pantothenic Acid (as Calcium Pantothenate or Racemic Calcium Pantothenate),Pyridoxine Hydrochloride,Riboflavin,and Thiamine Hydrochloride or Thiamine Mononitrate;and one mineral or more,furnishing one or more of the following elements in ionizable form:calcium,chromium,copper,fluorine,iodine,iron,magnesium,manganese,molybdenum,phosphorus,potassium,selenium,and zinc,derived from substances generally recognized as safe.Capsules contain not less than 90.0percent and not more than 165.0percent of the labeled amounts of vitamin A(C20H30O)as retinol or esters of retinol in the form of retinyl acetate (C22H32O2)or retinyl palmitate (C36H60O2),vitamin Das cholecalciferol (C27H44O)or ergocalciferol (C28H44O),vitamin Eas alpha tocopherol (C29H50O2)or alpha tocopheryl acetate (C31H52O3)or alpha tocopheryl acid succinate (C33H54O5),phytonadione (C31H46O2),and beta carotene (C40H56);and not less than 90.0percent and not more than 150.0percent of the labeled amounts of ascorbic acid (C6H8O6)or its salts as calcium ascorbate (C12H14CaO12·2H2O)or sodium ascorbate (C6H7NaO6),biotin (C10H16N2O3S),cyanocobalamin (C63H88CoN14O14P),folic acid (C19H19N7O6),niacin (C6H5NO2)or niacinamide (C6H6N2O),dexpanthenol (C9H19NO4)or panthenol (C9H19NO4),calcium pantothenate (C18H32CaN2O10),pyridoxine hydrochloride (C8H11NO3·HCl),riboflavin (C17H20N4O6),and thiamine (C12H17ClN4OS)as thiamine hydrochloride or thiamine mononitrate;and not less than 90.0percent and not more than 125.0percent of the labeled amount of calcium (Ca),copper (Cu),iron (Fe),magnesium (Mg),manganese (Mn),phosphorus (P),potassium (K),and zinc (Zn);and not less than 90.0percent and not more than 125.0percent of the labeled amounts of chromium (Cr),fluorine (F),iodine (I),molybdenum (Mo),and selenium (Se).
They may contain other labeled added substances that are generally recognized as safe,in amounts that are unobjectionable.
Packaging and storage— Preserve in tight,light-resistant containers.
Labeling*— The label states that the product is Oil-and Water-Soluble Vitamins with Minerals Capsules.The label also states the quantity of each vitamin and mineral per dosage unit and where necessary the chemical form in which a vitamin is present and also states the salt form of the mineral used as the source of each element.Where the product contains vitamin E,the label indicates whether it is the d-or dl-form.Where more than one Assaymethod is given for a particular vitamin or mineral,the labeling states with which Assaymethod the product complies only if Method 1is not used.
Microbial enumeration á2021ñ— The total aerobic microbial count does not exceed 3000cfu per g,and the combined molds and yeasts count does not exceed 300cfu per g.Capsules also meet the requirements of the tests for absence of Salmonellaspecies,Escherichia coli,and Staphylococcus aureus.
Disintegration and dissolution á2040ñ: meet the requirements for Dissolution.
Weight variation á2091ñ: meet the requirements.
NOTE—In the following assays,where more than one Assaymethod is given for an individual ingredient,compliance may be determined by following any one of the specified methods,the method used being stated in the labeling only if Method 1is not used.
Assay for vitamin A,Method 1— Proceed as directed in the Assay for vitamin A,Method 1under Oil-and Water-Soluble Vitamins with Minerals Tablets,except to read Capsules in place of Tablets and to use the following Assay preparation.
Assay preparation— Transfer the contents of not fewer than 20Capsules to a suitable container,mix,and weigh.Transfer an accurately weighed portion of the mixture,equivalent to 5Capsules,to a container having a polytef-lined screw cap.[NOTE—For hard gelatin Capsules,remove,as completely as possible,the contents of not fewer than 20Capsules by cutting open the Capsule shells,using a sharp blade if necessary,transferring the shells and their contents to a suitable container,and triturating to a homogeneous mass.Transfer an accurately weighed portion of the mass,equivalent to 5Capsules,to a container having a polytef-lined screw cap.]Add 10mLof dimethyl sulfoxide and 15mLof n-hexane,and shake for 45minutes on a wrist-action shaker in a water bath maintained at 60.[NOTE—Set up the wrist-action shaker to ensure that the contents of the container are mixed vigorously and thoroughly.]Centrifuge at 3000rpm for 10minutes,and transfer the hexane layer by means of a pipet to a 100-mLvolumetric flask.Add 15mLof n-hexane to the dimethyl sulfoxide layer,shake thoroughly for 5minutes,and transfer the hexane layer by means of a pipet to the 100-mLvolumetric flask.Repeat this extraction with three additional 15-mLportions of n-hexane.Dilute the extracts in the volumetric flask with n-hexane to volume,and mix.Quantitatively dilute a 10-mLvolume of this solution with n-hexane to obtain a solution having a final concentration of about 15µg of retinol per mL.Retain the remaining solution for use in the assays for vitamin D,vitamin E,and phytonadione.
Assay for vitamin A,Method 2— Proceed as directed in the Assay for vitamin A,Method 2under Oil-and Water-Soluble Vitamins with Minerals Tablets,except to read Capsules in place of Tablets and to use the following Assay preparation.
Assay preparation— [NOTE—This preparation is suitable for the determination of vitamin A,vitamin D,and vitamin E,when present in the formulation.]Accurately weigh not fewer than 20Capsules in a tared weighing bottle.Using a sharp blade if necessary,carefully open the Capsules,without loss of shell material,and transfer the contents to a 100-mLbeaker.Remove any contents adhering to the empty shells by washing with several portions of ether.Discard the washings,and dry the Capsule shells with the aid of a current of dry air.Weigh the empty Capsule shells in the tared weighing bottle,and calculate the net weight of the Capsule contents.Transfer an accurately weighed portion of the Capsule contents,equivalent to about 30µg of cholecalciferol or ergocalciferol (vitamin D),to a container having a polytef-lined screw cap.If vitamin Dis not present in the formulation,use a portion equivalent to about 90mg of vitamin E.If vitamin Eis not present in the formulation,use a portion equivalent to about 2.5mg of vitamin A.Proceed as directed for Assay preparationin the Assay for vitamin A,Method 2under Oil-and Water-Soluble Vitamins with Minerals Tablets,beginning with “Add about 0.5g of sodium bicarbonate.”
Assay for vitamin A,Method 3— Proceed as directed in the Assay for vitamin A,Method 3under Oil-and Water-Soluble Vitamins with Minerals Tablets,except to read Capsules in place of Tablets and to use the following Assay preparation.
Assay preparation— Accurately weigh not fewer than 20Capsules in a tared weighing bottle.Using a sharp blade if necessary,carefully open the Capsules,without loss of shell material,and transfer the contents to a 100-mLbeaker.Remove any contents adhering to the empty shells by washing with several portions of ether.Discard the washings,and dry the Capsule shells with the aid of a current of dry air.Weigh the empty Capsule shells in the tared weighing bottle,and calculate the net weight of the Capsule contents.Transfer an accurately weighed portion of the Capsule contents,equivalent to about 1.5mg of retinyl acetate,to a stoppered 125-mLflask,and proceed as directed for Assay preparationin the Assay for vitamin A,Method 3under Oil-and Water-Soluble Vitamins with Minerals Tablets,beginning with “Add 5mLof water.”
Assay for cholecalciferol or ergocalciferol (vitamin D),Method 1— Proceed as directed in the Assay for cholecalciferol or ergocalciferol(vitamin D),Method 1under Oil-and Water-Soluble Vitamins with Minerals Tablets,except to read Capsules in place of Tablets and to use the following Assay preparation.
Assay preparation— Transfer an accurately measured volume of the solution retained as specified in the directions for Assay preparationin the Assay for vitamin A,Method 1to a suitable container,and evaporate,if necessary,in vacuum at room temperature to obtain a solution having a concentration of about 2µg of cholecalciferol or ergocalciferol per mL.
Assay for cholecalciferol or ergocalciferol (vitamin D),Method 2— Proceed as directed in the Assay for cholecalciferol or ergocalciferol(vitamin D),Method 2under Oil-and Water-Soluble Vitamins with Minerals Tablets,except to read Capsules in place of Tablets and to use the following Assay preparation.
Assay preparation— Proceed as directed for Assay preparationin the Assay for vitamin A,Method 2.
Assay for cholecalciferol or ergocalciferol (vitamin D),Method 3— Proceed as directed in the Assay for cholecalciferol or ergocalciferol(vitamin D),Method 3under Oil-and Water-Soluble Vitamins with Minerals Tablets,except to read Capsules in place of Tablets and to use the following Assay preparation.
Assay preparation— Proceed as directed for Assay preparationin the Assay for vitamin A,Method 3,through “calculate the net weight of the Capsule contents.”Transfer an accurately weighed portion of the Capsule contents,equivalent to about 10µg of ergocalciferol or cholecalciferol,to a stoppered 125-mLflask,and proceed as directed for Standard preparation,beginning with “Add 15.0mLof water and 15.0mLof Potassium hydroxide solution.”
Assay for vitamin E,Method 1— Proceed as directed in the Assay for vitamin E,Method 1under Oil-and Water-Soluble Vitamins with Minerals Tablets,except to read Capsules in place of Tablets and to use the following Assay preparation.
Assay preparation— Transfer not less than 20mL,accurately measured,of the solution retained as specified in the directions for Assay preparationin the Assay for vitamin A,Method 1to a suitable container,and evaporate if necessary,in vacuum at room temperature to dryness.Transfer the contents of the flask to a suitable volumetric flask with the aid of methanol,and dilute with methanol to volume to obtain a solution having a concentration of about 2mg of alpha tocopherol,alpha tocopheryl acetate,or alpha tocopheryl acid succinate per mL.
Assay for vitamin E,Method 2— Proceed as directed in the Assay for vitamin E,Method 2under Oil-and Water-Soluble Vitamins with Minerals Tablets,except to read Capsules in place of Tablets and to use the following 3N Methanolic sulfuric acid solution,Sodium ascorbate-pyrogallol solution,Lecithin solution,and Assay preparation.
3N Methanolic sulfuric acid solution,Sodium ascorbate-pyrogallol solution,and Lecithin solution— Proceed as directed in the Assay for vitamin A,Method 2under Oil-and Water-Soluble Vitamins with Minerals Tablets.
Assay preparation— Proceed as directed for Assay preparationin the Assay for vitamin A,Method 2,through “calculate the net weight of the Capsule contents.”Transfer an accurately weighed portion of the Capsule contents,equivalent to about 55mg of vitamin E,to a container having a polytef-lined screw cap.Add about 0.5g of sodium bicarbonate,1.5mLof Lecithin solution,and 12.5mLof 2,2,4-trimethylpentane,and disperse on a vortex mixer.Add 6mLof Sodium ascorbate-pyrogallol solution,shake slowly,and allow the solution to degas.Continue shaking until the evolution of gas has ceased,and then shake for an additional 12minutes.Add 6mLof dimethyl sulfoxide,mix on a vortex mixer to form a suspension,and shake for 12minutes.Add 6mLof 3N Methanolic sulfuric acid solution,mix on a vortex mixer to form a suspension,and shake for 12minutes.Add 12.5mLof 2,2,4-trimethylpentane,mix on a vortex mixer to form a suspension,and shake for 10minutes.Centrifuge for about 10minutes to break up the emulsion and to clarify the supernatant layer.Transfer an accurately measured volume of the supernatant 2,2,4-trimethylpentane layer to a suitable volumetric flask,the volume of the specimen withdrawn from the 2,2,4-trimethylpentane layer and the size of the volumetric flask being such that the final concentration of the Assay preparationis equivalent to that of the Standard preparation.Evaporate nearly to dryness,add several mLof methanol,and evaporate the remaining 2,2,4-trimethylpentane.Dilute with methanol to volume,and mix.
Assay for vitamin E,Method 3— Proceed as directed in the Assay for vitamin E,Method 3under Oil-and Water-Soluble Vitamins with Minerals Tablets,except to read Capsules in place of Tablets and to use the following Assay preparation.
Assay preparation— Proceed as directed for Assay preparationin the Assay for vitamin A,Method 3,through “calculate the net weight of the Capsule contents.”Transfer an accurately weighed portion of the Capsule contents,equivalent to about 8.0mg of alpha tocopherol,to a glass-stoppered conical flask,and proceed as directed for Assay preparationin the Assay for vitamin E,Method 3under Oil-and Water-Soluble Vitamins with Minerals Tablets,beginning with “Add 25.0mLof water.”
Assay for phytonadione— Proceed as directed in the Assay for phytonadioneunder Oil-and Water-Soluble Vitamins with Minerals Tablets,except to read Capsules in place of Tablets and to use the following Assay preparation.
Assay preparation— Transfer not less than 20mL,accurately measured,of the solution retained as specified in the directions for Assay preparationin the Assay for vitamin A,Method 1to a suitable container,and evaporate,if necessary,in vacuum at room temperature to dryness.Transfer the contents of the flask to a suitable volumetric flask with the aid of methanol,and dilute with methanol to volume to obtain a solution having a concentration of about 20µg of phytonadione per mL.
Assay for beta carotene— Proceed as directed in the Assay for beta caroteneunder Oil-and Water-Soluble Vitamins with Minerals Tablets,except to read Capsules in place of Tablets and to use one of the following Assay preparations.
Assay preparation 1(for preparations containing beta carotene in oil solutions)— Proceed as directed in the Assay for vitamin A,Method 1,except to use cyclohexane instead of n-hexane as the extraction solvent and to dilute the filtered extracts quantitatively,and stepwise if necessary,with cyclohexane to obtain a solution having a concentration of about 2µg of beta carotene per mL.
Assay preparation 2(for preparations containing beta carotene in dry powder)— Remove the contents of not fewer than 20Capsules by cutting open the Capsules,using a sharp blade if necessary.Mix,and determine the weight of the contents.Transfer an accurately weighed quantity of the Capsule contents,equivalent to about 2mg of beta carotene,to a 500-mLsaponification flask,and proceed as directed for Assay preparationin the Assay for beta caroteneunder Oil-and Water-Soluble Vitamins with Minerals Tablets,beginning with “Add 100mLof alcohol.”
Assay for ascorbic acid,Method 1— Accurately weigh not fewer than 20Capsules in a tared weighing bottle.Using a sharp blade if necessary,carefully cut open the Capsules,without the loss of shell material,and transfer the contents to a 100-mLbeaker.Remove any contents adhering to the empty shells by washing if necessary with several portions of ether.Discard the washings,and dry the Capsule shells with the aid of a current of dry air until the odor of ether is no longer perceptible.Weigh the empty Capsule shells in the tared weighing bottle,and calculate the average net weight per Capsule.Transfer an accurately weighed portion of the Capsule contents,equivalent to about 100mg of ascorbic acid,to a 200-mLvolumetric flask,and proceed as directed in the Assay for ascorbic acid,Method 1under Oil-and Water-Soluble Vitamins with Minerals Tablets,beginning with “add about 75mLof metaphosphoric-acetic acids TS”and calculating the content of ascorbic acid in each Capsule.
Assay for ascorbic acid,Method 2— Proceed as directed in the Assay for Ascorbic Acidunder Automated Methods of Analysis á16ñ.
Assay for calcium ascorbate,Method 1— Proceed as directed in the Assay for ascorbic acid,Method 1.
Assay for calcium ascorbate,Method 2— Proceed as directed in the Assay for Ascorbic Acidunder Automated Methods of Analysis á16ñ.
Assay for sodium ascorbate,Method 1— Proceed as directed in the Assay for ascorbic acid,Method 1.
Assay for sodium ascorbate,Method 2— Proceed as directed in the Assay for Ascorbic Acidunder Automated Methods of Analysis á16ñ.
Assay for biotin,Method 1— Proceed as directed in the Assay for biotin,Method 1under Oil-and Water-Soluble Vitamins with Minerals Tablets,except to read Capsules in place of Tablets and to use the following Assay preparation.
Assay preparation— Proceed as directed in the Assay for ascorbic acid,Method 1,through “calculate the average net weight per Capsule.”Transfer an accurately weighed portion of the Capsule contents,equivalent to about 1mg of biotin,to a 200-mLvolumetric flask.Add 3mLof dimethyl sulfoxide,and swirl to wet the contents.Place the flask in a water bath at 60to 70for 5minutes.Sonicate for 5minutes,dilute with water to volume,mix,and filter.
Assay for biotin,Method 2— Proceed as directed in the Assay for biotin,Method 2under Oil-and Water-Soluble Vitamins with Minerals Tablets,except to read Capsules in place of Tablets and to use the following Assay preparation.
Assay preparation— Proceed as directed in the Assay for ascorbic acid,Method 1,through “calculate the average net weight per Capsule.”Transfer an accurately weighed portion of the Capsule contents,equivalent to about 100µg of biotin,to a 200-mLvolumetric flask,add 3mLof 50percent alcohol,and swirl to wet the contents.Heat the flask in a water bath at 60to 70for 5minutes.Sonicate for 5minutes,dilute with diluted alcohol to volume,mix,and filter.Dilute an accurately measured volume of the filtrate quantitatively,and stepwise if necessary,with water to obtain a solution having a concentration of about 0.1ng per mL.
Assay for cyanocobalamin,Method 1— Proceed as directed in the Assay for cyanocobalamin,Method 1under Oil-and Water-Soluble Vitamins with Minerals Tablets,except to read Capsules in place of Tablets and to use the following Assay preparation.
Assay preparation— Proceed as directed in the Assay for ascorbic acid,Method 1,through “calculate the average net weight per Capsule.”Transfer an accurately weighed portion of the Capsule contents,equivalent to about 100µg of cyanocobalamin,to a 250-mLflask.Quantitatively add 100.0mLof water,and carefully extract for 2minutes.Filter about 10mLof the extract,and use the clear filtrate.
Assay for cyanocobalamin,Method 2— Proceed as directed in the Assay for cyanocobalamin,Method 2under Oil-and Water-Soluble Vitamins with Minerals Tablets,except to read Capsules in place of Tablets and to use the following Assay preparation.
Assay preparation— Proceed as directed in the Assay for ascorbic acid,Method 1,through “calculate the average net weight per Capsule.”Transfer an accurately weighed portion of the Capsule contents,equivalent to about 1.0µg of cyanocobalamin,to an appropriate vessel containing,for each g of Capsule contents taken,25mLof an aqueous extracting solution prepared just prior to use to contain,in each 100mL,1.29g of dibasic sodium phosphate,1.1g of anhydrous citric acid,and 1.0g of sodium metabisulfite.Autoclave the mixture at 121for 10minutes.Allow any undissolved particles of the extract to settle,and filter or centrifuge,if necessary.Dilute an aliquot of the clear solution with water to obtain a final solution containing vitamin B12activity approximately equivalent to that of the Standard preparation.
Assay for folic acid,Method 1— Proceed as directed in the Assay for folic acid,Method 1under Oil-and Water-Soluble Vitamins with Minerals Tablets,except to read Capsules in place of Tablets and to use the following Assay preparation.
Assay preparation— Proceed as directed in the Assay for ascorbic acid,Method 1,through “calculate the average net weight per Capsule.”Transfer an accurately weighed portion of the Capsule contents,equivalent to about 0.4mg of folic acid,to a 50-mLcentrifuge tube.Add 25.0mLof Internal standard preparation,insert a stopper,shake by mechanical means for 10minutes,and centrifuge.Filter a portion of the clear supernatant,and use the filtrate.
Assay for folic acid,Method 2— Proceed as directed in the Assay for folic acid,Method 2under Oil-and Water-Soluble Vitamins with Minerals Tablets,except to read Capsules in place of Tablets and to use the following Assay preparation.
Assay preparation— Proceed as directed for Assay preparationin the Assay for vitamin A,Method 3,through “calculate the net weight of the Capsule contents.”Transfer an accurately weighed portion of the Capsule contents,equivalent to about 0.3mg of folic acid,to a stoppered 125-mLflask.Add 10.0mLof methanol and 35.0mLof Reagent,insert the stopper,shake for 15minutes in a water bath maintained at 60,and cool.Filter,discarding the first few mLof the filtrate.
Assay for dexpanthenol or panthenol— Proceed as directed in the Assay for dexpanthenol or panthenolunder Water-Soluble Vitamins Capsules.
Assay for calcium pantothenate,Method 1— Proceed as directed in the Assay for calcium pantothenate,Method 1under Oil-and Water-Soluble Vitamins with Minerals Tablets,except to read Capsules in place of Tablets and to use the following Assay preparation.
Assay preparation— Proceed as directed in the Assay for ascorbic acid,Method 1,through “calculate the average net weight per Capsule.”Transfer an accurately weighed portion of the Capsule contents,equivalent to about 15mg of calcium pantothenate,to a centrifuge tube.Transfer 25.0mLof the Internal standard preparationto the centrifuge tube,and shake vigorously for 10minutes.Centrifuge,filter,and use the clear filtrate.
Assay for calcium pantothenate,Method 2— Proceed as directed in the Assay for calcium pantothenate,Method 2under Oil-and Water-Soluble Vitamins with Minerals Tablets,except to read Capsules in place of Tablets and to use the following Assay preparation.
Assay preparation— Proceed as directed in the Assay for ascorbic acid,Method 1,through “calculate the average net weight per Capsule.”Transfer an accurately weighed portion of the Capsule contents,equivalent to about 50mg of calcium pantothenate,to a 1000-mLvolumetric flask containing 500mLof water.Add 10mLof 0.2Nacetic acid and 100mLof sodium acetate solution (1in 60),dilute with water to volume,and filter.Dilute an accurately measured volume of this solution quantitatively,and stepwise if necessary,to obtain a solution having approximately the same concentration as that of the Standard preparation.
Assay for calcium pantothenate,Method 3— Proceed as directed in the Assay for calcium pantothenate,Method 3under Oil-and Water-Soluble Vitamins with Minerals Tablets,except to read Capsules in place of Tablets and to use the following Assay preparation.
Assay preparation— Proceed as directed for Assay preparationin the Assay for folic acid,Method 2,through “calculate the net weight of the Capsule contents.”Transfer an accurately weighed portion of the Capsule contents,equivalent to about 10mg of calcium pantothenate,to a 250-mLvolumetric flask,add 10mLof methanol,and swirl the flask to disperse the Capsules contents.Dilute with water to volume,mix,and filter.
Assay for niacin or niacinamide,pyridoxine hydrochloride,riboflavin,and thiamine,Method 1— Proceed as directed in the Assay for niacin or niacinamide,pyridoxine hydrochloride,riboflavin,and thiamine,Method 1under Oil-and Water-Soluble Vitamins with Minerals Tablets,except to read Capsules in place of Tablets and to use the following Assay preparation.
Assay preparation— Proceed as directed in the Assay for ascorbic acid,Method 1,through “calculate the average net weight per Capsule.”Transfer an accurately weighed portion of the Capsule contents,equivalent to about 10mg of niacinamide and 2.5mg each of pyridoxine hydrochloride,riboflavin,and thiamine hydrochloride,to a 50-mLcentrifuge tube.Proceed as directed for Assay preparationin the Assay for niacin or niacinamide,pyridoxine hydrochloride,riboflavin,and thiamine,Method 1under Oil-and Water-Soluble Vitamins with Minerals Tablets,beginning with “Add 25.0mLof Diluting solution”.
Assay for niacin,Method 2— Proceed as directed in the Assay for niacin,Method 2under Oil-and Water-Soluble Vitamins with Minerals Tablets,except to read Capsules in place of Tablets and to use the following Assay preparation.
Assay preparation— [NOTE—This preparation is suitable for the determination of niacin or niacinamide,pyridoxine hydrochloride,and riboflavin,when present in the formulation.]Accurately weigh not fewer than 20Capsules in a tared weighing bottle.Using a sharp blade if necessary,carefully open the Capsules,without loss of shell material,and transfer the contents to a beaker.Remove any contents adhering to the shells by washing with several portions of ether.Discard the washings,and dry the Capsule shells with the aid of a current of dry air.Weigh the empty Capsule shells in the tared weighing bottle,and calculate the net weight of the Capsule contents.Transfer an accurately weighed portion of the Capsule contents,equivalent to about 2mg of riboflavin,to a 250-mLvolumetric flask.If riboflavin is not present in the formulation,use a portion equivalent to about 2mg of pyridoxine.If pyridoxine is not present in the formulation,use a portion equivalent to about 20mg of niacin or niacinamide.Add 100.0mLof Extraction solvent,and mix for 20minutes,using a wrist-action shaker.Immerse the flask in a water bath maintained at 70to 75,and heat for 20minutes.Mix on a vortex mixer for 30seconds,cool to room temperature,and filter.Use the clear filtrate.
Assay for niacinamide,Method 2— Proceed as directed in the Assay for niacinamide,Method 2under Oil-and Water-Soluble Vitamins with Minerals Tablets,except to read Capsules in place of Tablets and to use the following Assay preparation.
Assay preparation— Prepare as directed for Assay preparationin the Assay for niacin,Method 2.
Assay for pyridoxine hydrochloride,Method 2— Proceed as directed in the Assay for pyridoxine hydrochloride,Method 2under Oil-and Water-Soluble Vitamins with Minerals Tablets,except to read Capsules in place of Tablets and to use the following Assay preparation.
Assay preparation— Prepare as directed for the Assay preparationin the Assay for niacin,Method 2.
Assay for riboflavin,Method 2— Proceed as directed in the Assay for riboflavin,Method 2under Oil-and Water-Soluble Vitamins with Minerals Tablets,except to read Capsules in place of Tablets and to use the following Assay preparation.
Assay preparation— Prepare as directed for Assay preparationin the Assay for niacin,Method 2.
Assay for thiamine,Method 2— Proceed as directed in the Assay for thiamine,Method 2under Oil-and Water-Soluble Vitamins with Minerals Tablets,except to read Capsules in place of Tablets and to use the following Assay preparation.
Assay preparation— Proceed as directed for Assay preparationin the Assay for vitamin A,Method 3,through “calculate the net weight of the Capsule contents.”Transfer an accurately weighed portion of the contents,equivalent to about 2mg of thiamine,to a 250-mLvolumetric flask.Add 100.0mLof 0.2Nhydrochloric acid,shake for 15minutes with a wrist-action shaker,and heat to boiling for 30minutes.Cool to room temperature,and filter.Use the clear filtrate.
Assay for niacin or niacinamide,pyridoxine hydrochloride,riboflavin,and thiamine,Method 3— Proceed as directed in the Assay for niacin or niacinamide,pyridoxine hydrochloride,riboflavin,and thiamine,Method 3under Oil-and Water-Soluble Vitamins with Minerals Tablets,except to read Capsules in place of Tablets and to use the following Assay preparation.
Assay preparation— Proceed as directed for Assay preparationin the Assay for vitamin A,Method 3,through “calculate the net weight of the Capsule contents.”Transfer an accurately weighed portion of the Capsule contents,equivalent to about 7.5mg of niacin or niacinamide,1.2mg of pyridoxine hydrochloride,0.4mg of riboflavin,and 1.2mg of thiamine hydrochloride,to a stoppered 125-mLflask.Add 10.0mLof a mixture of methanol and glacial acetic acid (9:1)and 30.0mLof a mixture of methanol and ethylene glycol (1:1).Insert the stopper,shake for 15minutes in a water bath maintained at 60,and cool.Filter,discarding the first few mLof the filtrate.
NOTE—Commercially available atomic absorption standard solutions for the minerals,where applicable,may be used where preparation of a standard stock solution is described in the following Assays.Use deionized water where water is specified.Where atomic absorption spectrophotometry is specified in the Assay,the Standard preparationsand the Assay preparationmay be diluted quantitatively with the solvent specified,if necessary,to yield solutions of suitable concentrations adaptable to the linear or working range of the instrument.
Assay for calcium— Proceed as directed in the Assay for calciumunder Oil-and Water-Soluble Vitamins with Minerals Tablets,except to read Capsules in place of Tablets and to use the following Polysorbate 80solutionand Assay preparation.
Polysorbate 80solution— Dilute 100mLof polysorbate 80with alcohol to 1000mL.
Assay preparation— Transfer 5Capsules to a 100-mLvolumetric flask.[NOTE—For hard gelatin Capsules,accurately weigh not fewer than 20Capsules.Using a sharp blade if necessary,carefully open the Capsules,without loss of shell material,and transfer the contents to a suitable container.Remove any contents adhering to the empty shells by washing with several portions of ether.Discard the washings,and allow the Capsule shells to dry.Weigh the empty Capsule shells,calculate the net weight of the Capsule contents,and transfer an accurately weighed portion of the Capsule contents,equivalent to about 5Capsules,to a 100-mLvolumetric flask.]Add about 15mLof water,10mLof 6Nhydrochloric acid,and about 1mLof Polysorbate 80solutionto the flask,and heat on a hot plate or steam bath,with intermittent swirling,until the Capsules are completely disintegrated or the contents are dissolved.Boil gently for an additional 15minutes.Cool,dilute with water to volume,and filter,discarding the first 5mLof the filtrate.Dilute this solution quantitatively,and stepwise if necessary,with 0.125Nhydrochloric acid to obtain a solution having a known concentration of about 2µg of calcium per mL,adding 1mLof Lanthanum chloride solutionper 100mLof the final volume.
Assay for chromium— Proceed as directed in the Assay for chromiumunder Oil-and Water-Soluble Vitamins with Minerals Tablets,except to read Capsules in place of Tablets and to use the following Polysorbate 80solutionand Assay preparation.
Polysorbate 80solution— Prepare as directed in the Assay for calcium.
Assay preparation— Proceed as directed for Assay preparationin the Assay for calcium,except to prepare the Assay preparationto contain about 1µg of chromium per mLand to omit the use of the Lanthanum chloride solution.
Assay for copper— Proceed as directed in the Assay for copperunder Oil-and Water-Soluble Vitamins with Minerals Tablets,except to read Capsules in place of Tablets and to use the following Polysorbate 80solutionand Assay preparation.
Polysorbate 80solution— Prepare as directed in the Assay for calcium.
Assay preparation— Proceed as directed for Assay preparationin the Assay for calcium,except to prepare the Assay preparationto contain about 2µg of copper per mLand to omit the use of the Lanthanum chloride solution.
Assay for fluoride,Method 1— Proceed as directed in the Assay for fluoride,Method 1under Oil-and Water-Soluble Vitamins with Minerals Tablets,except to read Capsules in place of Tablets and to use the following Assay preparation.
Assay preparation— Remove the contents of a counted number of Capsules by cutting open the Capsules,using a sharp blade if necessary.Mix,and determine the weight of the contents.Transfer an accurately weighed quantity of the contents,equivalent to about 200µg of fluoride,to a 100-mLvolumetric flask.Add 10.0mLof 1Nhydrochloric acid,25.0mLof 3M Sodium acetate solution,and 25.0mLof Sodium citrate solution,dilute with water to volume,and mix.
Assay for fluoride,Method 2— Proceed as directed in the Assay for fluoride,Method 2,under Oil-and Water-Soluble Vitamins with Minerals Tablets,except to read Capsules in place of Tablets and to use the following Assay preparation.
Assay preparation— Accurately weigh not fewer than 20Capsules in a tared weighing bottle.Using a sharp blade if necessary,carefully open the Capsules,without loss of shell material,and transfer the contents to a 100-mLcontainer.If necessary,remove any contents adhering to the empty shells by washing with several portions of ether.Discard the washings,and dry the Capsule shells with the aid of a current of dry air.Weigh the empty Capsule shells in the tared weighing bottle,and calculate the net weight of the Capsule contents.Transfer an accurately weighed portion of the Capsule contents,equivalent to about 1mg of fluoride,to a 100-mLvolumetric flask.Proceed as directed for Assay preparationin the Assay for fluoride,Method 2under Oil-and Water-Soluble Vitamins with Minerals Tablets,beginning with “add 15mLof water”.
Assay for iodide,Method 1—
Bromine water— To about 20mLof bromine in a glass-stoppered bottle,add 100mLof water.Insert the stopper into the bottle,and shake.Allow to stand for 30minutes,and use the supernatant layer.
Procedure— Remove the contents of a counted number of Capsules by cutting open the Capsules,using a sharp blade if necessary.Mix,and determine the weight of the contents.Transfer an accurately weighed quantity of the contents,equivalent to about 3mg of iodine,to a nickel crucible.Proceed as directed for Procedurein the Assay for iodide,Method 1under Oil-and Water-Soluble Vitamins with Minerals Tablets,beginning with “Add 5g of sodium carbonate.”
Assay for iodide,Method 2— Proceed as directed in the Assay for Iodideunder Automated Methods of Analysis á16ñ.
Assay for iron— Proceed as directed in the Assay for ironunder Oil-and Water-Soluble Vitamins with Minerals Tablets,except to read Capsules in place of Tablets and to use the following Polysorbate 80solutionand Assay preparation.
Polysorbate 80solution— Prepare as directed in the Assay for calcium.
Assay preparation— Proceed as directed for Assay preparationin the Assay for calcium,except to prepare the Assay preparationto contain about 5µg of iron per mLand to omit the use of the Lanthanum chloride solution.
Assay for magnesium— Proceed as directed in the Assay for magnesiumunder Oil-and Water-Soluble Vitamins with Minerals Tablets,except to read Capsules in place of Tablets and to use the following Polysorbate 80solutionand Assay preparation.
Polysorbate 80solution— Prepare as directed in the Assay for calcium.
Assay preparation— Proceed as directed for Assay preparationin the Assay for calcium,except to prepare the Assay preparationto contain about 0.4µg of magnesium per mL.
Assay for manganese— Proceed as directed in the Assay for manganeseunder Oil-and Water-Soluble Vitamins with Minerals Tablets,except to read Capsules in place of Tablets and to use the following Polysorbate 80solutionand Assay preparation.
Polysorbate 80solution— Prepare as directed in the Assay for calcium.
Assay preparation— Proceed as directed for Assay preparationin the Assay for calcium,except to prepare the Assay preparationto contain about 1µg of manganese per mLand to omit the use of the Lanthanum chloride solution.
Assay for molybdenum,Method 1— Proceed as directed in the Assay for molybdenum,Method 1under Oil-and Water-Soluble Vitamins with Minerals Tablets,except to read Capsules in place of Tablets and to use the following Polysorbate 80solutionand Assay preparation.
Polysorbate 80solution— Prepare as directed in the Assay for calcium.
Assay preparation— Proceed as directed for Assay preparationin the Assay for calcium,except to take a number of Capsules or a portion of Capsule contents equivalent to about 1000µg of molybdenum and to make appropriate dilutions to obtain a final concentration of about 10µg of molybdenum per mL,omitting the addition of the Lanthanum chloride solution.
Assay for molybdenum,Method 2— Proceed as directed in the Assay for molybdenum,Method 2under Oil-and Water-Soluble Vitamins with Minerals Tablets,except to read Capsules in place of Tablets and to use the following Procedure.
Procedure— Remove the contents of a counted number of Capsules by cutting open the Capsules,using a sharp blade if necessary.Mix,and determine the weight of the contents.Transfer an accurately weighed quantity of the Capsule contents,equivalent to about 40µg of molybdenum,to a 200-mLbeaker,marked I.Proceed as directed for Procedurein the Assay for molybdenum,Method 2under Oil-and Water-Soluble Vitamins with Minerals Tablets,beginning with “To another 200-mLbeaker”,except to read Capsules in place of Tablets.
Assay for phosphorus— Proceed as directed in the Assay for phosphorusunder Oil-and Water-Soluble Vitamins with Minerals Tablets,except to read Capsules in place of Tablets and to use the following Assay preparation.
Assay preparation— Remove the contents of a counted number of Capsules by cutting open the Capsules,using a sharp blade if necessary.Mix,and determine the weight of the contents.Transfer an accurately weighed quantity of the Capsule contents,equivalent to about 100mg of phosphorus,to a suitable flask.Proceed as directed for Assay preparationin the Assay for phosphorusunder Oil-and Water-Soluble Vitamins with Minerals Tablets,beginning with “add 25mLof nitric acid”.
Assay for potassium— Proceed as directed in the Assay for potassiumunder Oil-and Water-Soluble Vitamins with Minerals Tablets,except to read Capsules in place of Tablets and to use the following Polysorbate 80solutionand Assay preparation.
Polysorbate 80solution— Prepare as directed in the Assay for calcium.
Assay preparation— Proceed as directed for Assay preparationin the Assay for calcium,except to prepare the Assay preparationto contain about 1µg of potassium per mLand to omit the use of the Lanthanum chloride solution.
Assay for selenium,Method 1— Proceed as directed in the Assay for selenium,Method 1under Oil-and Water-Soluble Vitamins with Minerals Tablets,except to read Capsules in place of Tablets and to use the following Assay preparation.
Assay preparation— Remove the contents of a counted number of Capsules by cutting open the Capsules,using a sharp blade if necessary.Mix,and determine the weight of the contents.Transfer an accurately weighed quantity of the Capsule contents,equivalent to about 1000µg of selenium,to a suitable flask.Proceed as directed for Assay preparationin the Assay for selenium,Method 1under Oil-and Water-Soluble Vitamins with Minerals Tablets,beginning with “and add about 12mLof nitric acid.”
Assay for selenium,Method 2— Proceed as directed in the Assay for selenium,Method 2under Oil-and Water-Soluble Vitamins with Minerals Tablets,except to read Capsules in place of Tablets and to use the following Assay preparation.
Assay preparation— Remove the contents of a counted number of Capsules by cutting open the Capsules,using a sharp blade if necessary.Mix,and determine the weight of the contents.Transfer an accurately weighed quantity of the Capsule contents,equivalent to about 20µg of selenium,to a suitable flask.Proceed as directed for Assay preparationin the Assay for selenium,Method 2under Oil-and Water-Soluble Vitamins with Minerals Tablets,beginning with “Add 10mLof nitric acid.”
Assay for zinc— Proceed as directed in the Assay for zincunder Oil-and Water-Soluble Vitamins with Minerals Tablets,except to read Capsules in place of Tablets and to use the following Polysorbate 80solutionand Assay preparation.
Polysorbate 80solution— Prepare as directed in the Assay for calcium.
Assay preparation— Proceed as directed for Assay preparationin the Assay for calcium,except to prepare the Assay preparationto contain about 2µg of zinc per mLand to omit the use of the Lanthanum chloride solution.
*  For information on USP Units of activity for vitamins and labeling in terms of units,see the footnote to the Labelingsection under Oil-and Water-Soluble Vitamins with Minerals Tablets.
Auxiliary Information— Staff Liaison:Lawrence Evans,III,Ph.D.,Scientist
Expert Committee:(DSN)Dietary Supplements:Non-Botanicals
USP28–NF23Page 2144
Pharmacopeial Forum:Volume No.28(5)Page 1544
Phone Number:1-301-816-8389