Saw Palmetto Capsules
»Saw Palmetto Capsules contain Saw Palmetto Extract.Capsules contain not less than 22.0percent of lauric acid and not more than 34.0percent of the labeled amount of Saw Palmetto Extract.The ratio of the concentrations of lauric acid to caprylic acid is not less than 8.5and not more than 17.5.The ratio of the concentrations of lauric acid to myristic acid is not less than 2.2and not more than 2.8.
Packaging and storage—
Preserve in tight,light-resistant containers.
Labeling—
The label states the Latin binomial and,following the official name,the name of article from which the Capsules were prepared.Label it to indicate the amount of Extract in mg per Capsule.
USP Reference standards á11ñ—
USP Methyl Caprate RS.USP Methyl Caproate RS.USP Methyl Caprylate RS.USP Methyl Laurate RS.USP Methyl Linoleate RS.USP Methyl Linolenate RS.USP Methyl Myristate RS.USP Methyl Oleate RS.USP Methyl Palmitate RS.USP Methyl Palmitoleate RS.USP Methyl Stearate RS.USPb-Sitosterol RS.
Identification—
A:
The retention times of the peaks for methyl caprate,methyl caproate,methyl caprylate,methyl laurate,methyl linoleate,methyl linolenate,methyl myristate,methyl oleate,methyl palmitate,methyl palmitoleate,and methyl stearate in the chromatogram of the Test solutioncorrespond to those in the chromatogram of the Standard solution,as obtained in the test for Content of lauric acid and the ratios of the concentrations of lauric acid to caprylic acid and lauric acid to myristic acid.
B:Presence of sterols—
Derivatizing reagent,Standard solution,and Chromatographic system—
Proceed as directed for Content of sterolsunder Saw Palmetto Extract.
Test solution—
Take a number of Capsules,equivalent to about 10g of extract,open the Capsules using a suitable cutting instrument,and transfer the shells and contents to a suitable container.Transfer about 5g,accurately weighed,to a 250-mLround-bottom flask,and evaporate in vacuum at a temperature not exceeding 50
.Add 50mLof a solution prepared by dissolving 130g of potassium hydroxide in 200mLof water and diluting with methanol to 1000mL.Attach a condenser,and reflux in a bath at 100until a clear solution is obtained.Reflux for an additional 10minutes,and cool by adding 50mLof water through the condenser.Quantitatively transfer to a separation funnel,rinsing the flask with a total of 50mLof water in small portions.Extract with 80mLof ether,shaking for 30seconds,and repeat twice.[NOTE—If an emulsion forms,it can be eliminated by adding small quantities of methanol.]Transfer the combined ether layers to a separation funnel,and wash with successive portions of 50mLof water until a neutral washing is obtained.[NOTE—If an emulsion forms,it can be eliminated by adding small quantities of methanol.]Pass the ether extract through filter paper containing anhydrous sodium sulfate,wash the filter with 30mLof ether,and evaporate to dryness in vacuum.Dissolve the residue in 2.0mLof chloroform.
.Add 50mLof a solution prepared by dissolving 130g of potassium hydroxide in 200mLof water and diluting with methanol to 1000mL.Attach a condenser,and reflux in a bath at 100until a clear solution is obtained.Reflux for an additional 10minutes,and cool by adding 50mLof water through the condenser.Quantitatively transfer to a separation funnel,rinsing the flask with a total of 50mLof water in small portions.Extract with 80mLof ether,shaking for 30seconds,and repeat twice.[NOTE—If an emulsion forms,it can be eliminated by adding small quantities of methanol.]Transfer the combined ether layers to a separation funnel,and wash with successive portions of 50mLof water until a neutral washing is obtained.[NOTE—If an emulsion forms,it can be eliminated by adding small quantities of methanol.]Pass the ether extract through filter paper containing anhydrous sodium sulfate,wash the filter with 30mLof ether,and evaporate to dryness in vacuum.Dissolve the residue in 2.0mLof chloroform.
Apply separately 200µLof this solution and 20µLof a solution of b-cholestanol in chloroform (1in 100)to a thin-layer chromatographic plate coated with 0.25-mm silica gel having an application zone that was previously dipped under 3cm of a solution prepared by dissolving 13g of potassium hydroxide in 20mLof water and diluted to 1000mLwith methanol.[NOTE—Allow the plate to dry,and heat it to 100for 1hour before use.The plate can be stored in a desiccator containing calcium chloride until the time of use.]Develop with a solvent consisting of a mixture of hexanes and ether (7:3)until the solvent front has moved 17to 19cm.Keep the chamber temperature between 15and 20.Dry the plate in a current of warm air,then spray with an alkaline solution of 2,7-dichlorofluorescein in alcohol (0.2in 100).Observe the plate under 366-nm wavelength light and identify the bands corresponding to the sterols by referring to the b-cholestanol spot.Scrape off these bands and transfer them to a test tube.Add 10mLof warm chloroform,and shake for 2minutes with the aid of several glass beads.Filter the chloroform solution,wash the filter with chloroform,and evaporate the combined filtrate and washings to dryness in vacuum.Dissolve the residue with some drops of anhydrous acetone and evaporate in vacuum.Dry the residue in an oven at 105for 15minutes.Dissolve the residue in 0.2mLof Derivatizing reagent,and allow to stand for not less than 15minutes at room temperature.
for 1hour before use.The plate can be stored in a desiccator containing calcium chloride until the time of use.]Develop with a solvent consisting of a mixture of hexanes and ether (7:3)until the solvent front has moved 17to 19cm.Keep the chamber temperature between 15and 20.Dry the plate in a current of warm air,then spray with an alkaline solution of 2,7-dichlorofluorescein in alcohol (0.2in 100).Observe the plate under 366-nm wavelength light and identify the bands corresponding to the sterols by referring to the b-cholestanol spot.Scrape off these bands and transfer them to a test tube.Add 10mLof warm chloroform,and shake for 2minutes with the aid of several glass beads.Filter the chloroform solution,wash the filter with chloroform,and evaporate the combined filtrate and washings to dryness in vacuum.Dissolve the residue with some drops of anhydrous acetone and evaporate in vacuum.Dry the residue in an oven at 105for 15minutes.Dissolve the residue in 0.2mLof Derivatizing reagent,and allow to stand for not less than 15minutes at room temperature.
Procedure—
Proceed as directed for Content of sterolsunder Saw Palmetto Extract.The chromatogram of the Test solutionexhibits peaks for campesterol,b-sitosterol,and stigmasterol,identified by their retention times relative to the b-sitosterol peak in the chromatogram of the Standard solution.
Microbial enumeration á2021ñ—
Capsules meet the requirements of the tests for absence of Salmonellaspecies,Escherichia coli,and Staphylococcus aureus.The total bacterial count does not exceed 10,000cfu per g,the total combined molds and yeasts count does not exceed 1000cfu per g,the coliform count does not exceed 100cfu per g,and the count for enterobacteria does not exceed 100cfu per g.
Rupture—
Medium:
simulated gastric fluid TS;500mL.
Apparatus 2:
50rpm (see Apparatusunder Disintegration and Dissolution of Dietary Supplements á2040ñ).
Time:
15minutes.
Procedure—
Place 1Capsule in each vessel,and allow the Capsule to sink to the bottom of the vessel before starting rotation of the blade.Record the time it takes for each Capsule shell to rupture.
Tolerances—
The requirements are met if all of the Capsules rupture in not more than 15minutes.If 1or 2of the Capsules rupture in more than 15but not more than 30minutes,repeat the test on 12additional Capsules.Not more than 2of the 18Capsules tested rupture in more than 15but not more than 30minutes.
Weight variation á2091ñ:
meet the requirements.
Content of lauric acid and the ratios of the concentrations of lauric acid to caprylic acid and lauric acid to myristic acid—
Internal standard solution,Standard solution,and Chromatographic system—
Proceed as directed for Content of fatty acidsunder Saw Palmetto.
Test solution—
Take a number of Capsules,equivalent to about 10g of extract,open the Capsules using a suitable cutting instrument,and transfer the shells and contents to a suitable container.Transfer about 100mg,accurately weighed,to a pressure-proof screw-capped vial,and add 3.0mLof a solution of sulfuric acid in methanol (5in 100).Heat in an oil bath at 100for 2hours,shaking from time to time.Allow to cool,and add 1.0mLof Internal standard solution,10.0mLof water,1g of sodium chloride,and 5mLof hexanes.Shake well,and allow the layers to separate completely.Use the hexanes layer.[NOTE—Store this solution in a refrigerator until use.]
for 2hours,shaking from time to time.Allow to cool,and add 1.0mLof Internal standard solution,10.0mLof water,1g of sodium chloride,and 5mLof hexanes.Shake well,and allow the layers to separate completely.Use the hexanes layer.[NOTE—Store this solution in a refrigerator until use.]
Procedure—
Proceed as directed for Content of fatty acidsunder Saw Palmetto.Identify the peaks of methyl caprate,methyl caproate,methyl caprylate,methyl laurate,methyl linoleate,methyl linolenate,methyl myristate,methyl oleate,methyl palmitate,methyl palmitoleate,and methyl stearate in the chromatogram of the Test solution.Calculate the quantity,in mg,of lauric acid,myristic acid,and caprylic acid in the portion of Capsules taken by the formula,
5C(RU/RS)(MA/ME),
in which the terms are as defined therein.Using these quantities,calculate the individual ratios of the concentration of lauric acid to caprylic acid and of lauric acid to myristic acid in the portion of Capsules taken.
Auxiliary Information—
Staff Liaison:Gabriel I.Giancaspro,Ph.D.,Senior Scientist and Latin American Specialist
Expert Committee:(DSB)Dietary Supplements:Botanicals
USP28–NF23Page 2129
Pharmacopeial Forum:Volume No.29(4)Page 1291
Phone Number:1-301-816-8343