Water-Soluble Vitamins Capsules
» Water-Soluble Vitamins Capsules contain two or more of the following water-soluble vitamins: Ascorbic Acid or its equivalent as Sodium Ascorbate or Calcium 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. Capsules contain not less than 90.0 percent and not more than 150.0 percent of the labeled amounts of ascorbic acid (C6H8O6), biotin (C10H16N2 O3S), cyanocobalamin (C63H88CoN14O14P), folic acid (C19H19N7O6), dexpanthenol (C9H19NO4) or panthenol (C9H19NO4), calcium pantothenate (C18H32CaN2O10), niacin (C6H5NO2) or niacinamide (C6H6N2O), pyridoxine hydrochloride (C8H11NO3·HCl), riboflavin (C17H20N4 O6), and thiamine (C12H17ClN4OS) as thiamine hydrochloride or thiamine mononitrate.
They do not contain any form of Beta Carotene or Vitamin A, D, E, or K. They do not contain any minerals for which nutritional value is claimed. They may contain other labeled added substances in amounts that are unobjectionable.
Packaging and storage
Preserve in tight, light-resistant containers.
Labeling
The label states that the product is Water-Soluble Vitamins Capsules. The label also states the quantity of each vitamin in terms of metric units per dosage unit and where necessary the salt form in which it is present. Where more than one Assay method is given for a particular vitamin, the labeling states which Assay method is used only if Method 1 is not used. Where products are labeled to contain panthenol, the label states the equivalent content of dexpanthenol.
USP Reference standards
![]() ![]() USP Biotin RS. USP Calcium Pantothenate RS. USP Cyanocobalamin RS. USP Dexpanthenol RS. USP Folic Acid RS. USP Niacin RS. USP Niacinamide RS. USP Pyridoxine Hydrochloride RS. USP Riboflavin RS. USP Thiamine Hydrochloride RS.
Microbial enumeration
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Weight variation
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noteIn the following Assays, where more than one Assay method 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 1 is not used.
Assay for ascorbic acid, Method 1
Accurately weigh not fewer than 20 Capsules in a tared weighing bottle. Using a sharp blade, carefully cut open the Capsules, without loss of shell material, and transfer the contents to a 100-mL beaker. 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 Capsule contents, equivalent to about 100 mg of ascorbic acid to a 200-mL volumetric flask. Proceed as directed in the Assay for ascorbic acid, Method 1 under Oil- and Water-Soluble Vitamins with Minerals Tablets, beginning with add about 75 mL of metaphosphoricacetic acids TS, and calculating the content of ascorbic acid in each Tablet.
Assay for ascorbic acid, Method 2
Proceed as directed in the Assay for Ascorbic Acid under Automated Methods of Analysis
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Assay for calcium ascorbate, Method 1
Proceed as directed under Assay for ascorbic acid, Method 1.
Assay for calcium ascorbate, Method 2
Proceed as directed in the Assay for Ascorbic Acid under Automated Methods of Analysis
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Assay for sodium ascorbate, Method 1
Proceed as directed under Assay for ascorbic acid, Method 1.
Assay for sodium ascorbate, Method 2
Proceed as directed in the Assay for Ascorbic Acid under Automated Methods of Analysis
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Assay for biotin, Method 1
Proceed as directed in the Assay for biotin, Method 1 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
Proceed as directed for Assay preparation in the Assay for biotin, Method 2 from the beginning through calculation of the net weight per Capsule. Transfer an accurately weighed portion of the Capsule contents, equivalent to about 1 mg of biotin, to a 200-mL volumetric flask. Add 3 mL of dimethyl sulfoxide, and swirl to wet the contents. Place the flask in a water bath at 60
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Assay for biotin, Method 2
Proceed as directed in the Assay for biotin, 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 20 Capsules in a tared weighing bottle. Using a sharp blade, carefully cut open the Capsules, without loss of shell material, and transfer the contents as completely as possible 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 until the odor of ether is no longer perceptible. Accurately 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 100 µg of biotin, to a 200-mL volumetric flask. Add 3 mL of 50 percent alcohol, and swirl to mix. Place the flask in a water bath at 60
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Assay for cyanocobalamin, Method 1
Proceed as directed in the Assay for cyanocobalamin, Method 1 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
Proceed as directed in the Assay for biotin, Method 2 from the beginning through calculation of the net weight per Capsule. Transfer an accurately weighed portion of the Capsule contents, equivalent to 100 µg of cyanocobalamin, to a 250-mL flask. Quantitatively add 100.0 mL of water, and carefully extract for 2 minutes. Filter about 10 mL of this solution, and use the clear filtrate.
Assay for cyanocobalamin, Method 2
Proceed as directed in the Assay for cyanocobalamin, 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
Proceed as directed for the Assay preparation under Assay for biotin, Method 2 from the beginning through calculation of the net weight per Capsule. Transfer an accurately weighed portion of the Capsule contents to an appropriate vessel containing, for each g or mL of Capsule contents taken, 25 mL of an aqueous extracting solution prepared just prior to use to contain, in each 100 mL, 1.29 g of dibasic sodium phosphate, 1.1 g of anhydrous citric acid, and 1.0 g of sodium metabisulfite. Autoclave the mixture at 121
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Assay for folic acid, Method 1
Proceed as directed in the Assay for folic acid, Method 1 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
Proceed as directed in the Assay preparation in the Assay for biotin, Method 2 from the beginning through calculation of the net weight per Capsule. Transfer an accurately weighed portion of the Capsule contents, equivalent to about 0.4 mg of folic acid, to a 50-mL amber-colored centrifuge tube. Add 25.0 mL of Internal standard preparation, insert the stopper, shake by mechanical means for 10 minutes, 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 2 under Water-Soluble Vitamins 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 biotin, Method 2 from the beginning through calculation of the average net weight per Capsule. Transfer an accurately weighed portion of the Capsule contents, equivalent to about 0.3 mg of folic acid, to a stoppered 125-mL flask. Add 10.0 mL of a mixture of methanol and glacial acetic acid (9:1) and 30.0 mL of a mixture of methanol and ethylene glycol (1:1). Insert the stopper, shake for 15 minutes in a water bath maintained at 60
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Assay for dexpanthenol or panthenol
[noteThe following procedure is applicable also to the determination of the dextrorotatory component of racemic panthenol in preparations containing panthenol.] Dehydrated mixtures yielding formulations similar to the media described herein may be used provided that, when constituted as directed, they have growth-promoting properties equal to or superior to those obtained with the media prepared as described herein.
Standard stock solution
Dissolve an accurately weighed quantity of USP Dexpanthenol RS in water, and dilute with water to obtain a solution having a known concentration of about 800 µg per mL. Store in a refrigerator, protected from light, and use within 30 days.
Standard preparation
On the day of the assay, dilute an accurately measured volume of Standard stock solution with water to obtain a solution having a known concentration of about 1.2 µg of dexpanthenol per mL.
Assay preparation
Accurately weigh not fewer than 30 Capsules in a tared weighing bottle. Using a sharp blade, carefully cut open the Capsules, without loss of shell material, and transfer the contents as completely as possible to a beaker. Remove any contents adhering to the empty Capsule 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 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. Dissolve an accurately weighed portion of the Capsule contents, equivalent to about 1.2 mg of dexpanthenol or 2.4 mg of panthenol, in 100.0 mL of water. Quantitatively dilute a portion of this solution with water to obtain a solution having a concentration of about 1.2 µg of dexpanthenol or 2.4 µg of panthenol per mL.
Acid-hydrolyzed casein solution
Mix 100 g of vitamin-free casein with 500 mL of 6 N hydrochloric acid, and reflux the mixture for 8 to 12 hours. Remove the hydrochloric acid from the mixture by distillation under reduced pressure until a thick paste remains. Redissolve the resulting paste in about 500 mL of water, adjust the solution with 1 N sodium hydroxide to a pH of 3.5 ± 0.1, and add water to make 1000 mL. Add 20 g of activated charcoal, stir for 1 hour, and filter. Repeat the treatment with activated charcoal. Store under toluene in a refrigerator at a temperature not below 10
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Cystine-tryptophane solution
Suspend 4.0 g of l-cystine and 1.0 g of l-tryptophane (or 2.0 g of d,l-tryptophane) in 700 to 800 mL of water, heat to 75 ± 5
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Adenine-guanine-uracil solution
Dissolve 200 mg each of adenine sulfate, guanine hydrochloride, and uracil, with the aid of heat, in 10 mL of 4 N hydrochloric acid, cool, add water to make 200 mL, and mix. Store under toluene in a refrigerator.
Polysorbate 80 solution
Dissolve 25 g of polysorbate 80 in alcohol to make 250 mL, and mix.
Riboflavin-thiamine hydrochloride-biotin solution
Prepare a solution of riboflavin, thiamine hydrochloride, and biotin in 0.02 N acetic acid containing 20 µg of riboflavin, 10 µg of thiamine hydrochloride, and 0.04 µg of biotin per mL. Store under toluene, protected from light, in a refrigerator.
p-Aminobenzoic acid-niacin-pyridoxine hydrochloride solution
Prepare a solution in neutral 25 percent alcohol containing 10 µg of p-aminobenzoic acid, 50 µg of niacin, and 40 µg of pyridoxine hydrochloride per mL. Store in a refrigerator.
Salt solution 1
Dissolve 25 g of monobasic potassium phosphate and 25 g of dibasic potassium phosphate in water to make 500 mL. Add 5 drops of hydrochloric acid, and mix. Store under toluene.
Salt solution 2
Dissolve 10 g of magnesium sulfate, 0.5 g of sodium chloride, 0.5 g of ferrous sulfate, and 0.5 g of manganese sulfate in water to make 500 mL. Add 5 drops of hydrochloric acid, and mix. Store under toluene.
Pyridoxal-calcium pantothenate solution
Dissolve 40 mg of pyridoxal hydrochloride and 375 µg of calcium pantothenate in 10 percent alcohol to make 200 mL, and mix. Store in a refrigerator, and use within 30 days.
Polysorbate 40-oleic acid solution
Dissolve 25 g of polysorbate 40 and 0.25 g of oleic acid in 20 percent alcohol to make 500 mL, and mix. Store in a refrigerator, and use within 30 days.
Modified pantothenate medium
Dissolve anhydrous dextrose and sodium acetate in the solutions previously mixed, and adjust with 1 N sodium hydroxide to a pH of 6.8. Finally, dilute with water to 250 mL, and mix.
Double-strength modified pantothenate medium
Prepare as directed under Modified pantothenate medium, but make the final dilution to 125 mL instead of 250 mL. Prepare fresh.
Stock culture of Pediococcus acidilactici
Dissolve in about 800 mL of water, with the aid of heat, 6.0 g of peptone, 4.0 g of pancreatic digest of casein, 3.0 g of yeast extract, 1.5 g of beef extract, 1.0 g of dextrose, and 15.0 g of agar. Adjust with 0.1 N sodium hydroxide or 0.1 N hydrochloric acid to a pH of between 6.5 and 6.6, dilute with water to 1000 mL, and mix. Add 10-mL portions of the solution to culture tubes, place caps on the tubes, and sterilize in an autoclave at 121
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Inoculum
Inoculate three 250-mL portions of Modified pantothenate medium from a stock culture slant, and incubate at 35
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Procedure
Prepare in triplicate a series of eight culture tubes by adding the following quantities of water to the tubes within a set: 5.0, 4.5, 4.0, 3.5, 3.0, 2.0, 1.0, and 0.0 mL. To these same tubes, and in the same order, add 0.0, 0.5, 1.0, 1.5, 2.0, 3.0, 4.0, and 5.0 mL of the Standard preparation.
Prepare in duplicate a series of five culture tubes by adding the following quantities of water to the tubes within a set: 4.0, 3.5, 3.0, 2.0, and 1.0 mL. To these same tubes, and in the same order, add 1.0, 1.5, 2.0, 3.0, and 4.0 mL of the Assay preparation.
Add 5.0 mL of Double-strength modified pantothenate medium to each tube, and mix. Cover the tubes with metal caps, and sterilize in an autoclave at 121
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Calculation
Draw a dose-response curve on arithmetic graph paper by plotting the average response, in percent transmittance, for each set of tubes of the standard curve against the standard level concentrations. The curve is drawn by connecting each adjacent pair of points with a straight line. From this standard curve, determine by interpolation the potency, in terms of dexpanthenol, of each tube containing portions of the Assay preparation. Divide the potency of each tube by the amount of the Assay preparation added to it, to obtain the individual responses. Calculate the mean response by averaging the individual responses that vary from their mean by not more than 15%, using not less than half the total number of tubes. Calculate the potency of the portion of the material taken for assay, in terms of dexpanthenol, by multiplying the mean response by the appropriate dilution factor.
Assay for calcium pantothenate, Method 1
Proceed as directed in the Assay for calcium pantothenate, Method 1 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
Proceed as directed in the Assay for biotin, Method 2 from the beginning through calculation of the net weight per Capsule. Transfer an accurately weighed portion of the Capsule contents, equivalent to about 15 mg of calcium pantothenate, to a centrifuge tube. Transfer 25.0 mL of Internal standard preparation to the centrifuge tube, and shake vigorously for 10 minutes. Centrifuge, filter, and use the clear filtrate.
Assay for calcium pantothenate, Method 2
Proceed as directed in the Assay for calcium pantothenate, 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
Proceed as directed in the Assay for biotin, Method 2 from the beginning through calculation of the net weight per Capsule. Transfer a portion of the Capsule contents, equivalent to about 50 mg of calcium pantothenate, to a 1000-mL volumetric flask containing 500 mL of water. Add 10 mL of 0.2 N acetic acid and 100 mL of sodium acetate solution (1 in 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 about 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 3, 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
Proceed as directed for Assay preparation in the Assay for biotin, Method 2 from the beginning through calculation of the average net weight per Capsule. Transfer an accurately weighed portion of the Capsule contents, equivalent to about 10 mg of calcium pantothenate, to a 250-mL volumetric flask, add 10 mL of 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 1 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
Proceed as directed in the Assay for biotin, Method 2 from the beginning through calculation of the average net weight of the Capsule. Transfer an accurately weighed portion of the Capsule contents, equivalent to 10 mg of niacinamide and 2.5 mg each of pyridoxine hydrochloride, riboflavin, and thiamine hydrochloride, to a 50-mL centrifuge tube, and add 25.0 mL of Diluting solution. Mix, using a vortex mixer, for 30 seconds to completely suspend the specimen. Immerse the centrifuge tube in a hot water bath maintained at 65
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Assay for niacin, Method 2
Proceed as directed in the Assay for niacin, 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
[noteThis preparation is suitable for the determination of niacin or niacinamide, pyridoxine hydrochloride, and riboflavin, when present in the formulation.] Accurately weigh not fewer than 20 Capsules 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 2 mg of riboflavin, to a 250-mL volumetric flask. If riboflavin is not present in the formulation, use a portion equivalent to about 2 mg of pyridoxine. If pyridoxine is not present in the formulation, use a portion equivalent to about 20 mg of niacin or niacinamide. Add 100.0 mL of Extraction solvent, and mix for 20 minutes, using a wrist-action shaker. Immerse the flask in a water bath maintained at 70
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Assay for niacinamide, Method 2
Proceed as directed in the Assay for niacinamide, 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
Prepare as directed for Assay preparation in the Assay for niacin, Method 2.
Assay for pyridoxine hydrochloride, Method 2
Proceed as directed in the Assay for pyridoxine hydrochloride, 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
Prepare as directed for Assay preparation in the Assay for niacin, Method 2.
Assay for riboflavin, Method 2
Proceed as directed in the Assay for riboflavin, 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 for thiamine, Method 2
Proceed as directed in the Assay for thiamine, 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
Prepare as directed for Assay preparation in the Assay for niacin, Method 2, through calculate the net weight of the Capsule contents. Transfer an accurately weighed portion of the contents, equivalent to about 2 mg of thiamine, to a 250-mL volumetric flask. Add 100.0 mL of 0.2 N hydrochloric acid, shake for 15 minutes with a wrist-action shaker, and heat to boiling for 30 minutes. 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 3 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
Proceed as directed for Assay preparation in the Assay for niacin, Method 2, through calculate the net weight of the Capsule contents. Transfer an accurately weighed portion of the Capsule contents, equivalent to about 7.5 mg of niacin or niacinamide, 1.2 mg of pyridoxine hydrochloride, 0.4 mg of riboflavin, and 1.2 mg of thiamine hydrochloride, to a stoppered 125-mL flask. Add 10.0 mL of a mixture of methanol and glacial acetic acid (9:1) and 30.0 mL of a mixture of methanol and ethylene glycol (1:1). Insert the stopper, shake for 15 minutes in a water bath maintained at 60
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*
ATCC No. 8042 is suitable.
Auxiliary Information
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Chromatographic Column
USP32NF27 Page 1124
Pharmacopeial Forum: Volume No. 27(1) Page 1989
Chromatographic columns text is not derived from, and not part of, USP 32 or NF 27.
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