NOTE—In the following Assays,where more than one Assaymethod is given for an individual ingredient,the requirements may be met by following any one of the specified methods,the method used being stated in the labeling only if Method 1is not used.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 concentrations of the Standard preparationsand the Assay preparationmay be modified to fit the linear or working range of the instrument.

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.

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.

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 preparation— Remove the contents of a counted number of Capsules by cutting open,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 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.”

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,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 in the Assay for iodide,Method 1under Oil-and Water-Soluble Vitamins with Minerals Tablets,beginning with “Add about 5g of sodium carbonate”.

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.

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.

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.

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.

Procedure— Remove the contents of a counted number of Capsules by cutting open,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 preparation— Remove the contents of a counted number of Capsules by cutting open,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 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”.

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 1µg of potassium per mLand to omit the use of the Lanthanum chloride solution.

Assay preparation— Remove the contents of a counted number of Capsules by cutting open,using a sharp blade if necessary.Mix,and determine the weight of the contents.Transfer an accurately weighed portion 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 preparation— Remove the contents of a counted number of Capsules by cutting open,using a sharp blade if necessary.Mix,and determine the weight of the contents.Transfer an accurately weighed portion of the 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.”

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.