Powdered Valerian Extract
»Powdered Valerian Extract is prepared from comminuted Valerian and with 70percent alcohol or other suitable solvents.It contains not less than 0.3percent of valerenic acid (C15H22O2).The ratio of the starting crude plant material to the Extract is between 4:1and 7:1.
Packaging and storage— Preserve in tight containers,store at controlled room temperature,and protect from moisture and light.
Labeling— The label states the official name of the article and states also the Latin binomial and the part of the plant from which the article was prepared.Label it to indicate the content of valerenic acid,the extracting solvent used for preparation,and the ratio of the starting crude plant material to the Extract.
Identification—
A: Thin-Layer Chromatographic Identification Test á201ñ—
Adsorbent: 0.5-mm layer of chromatographic silica gel mixture.
Test solution— Dissolve about 0.2g of Extract,accurately weighed,in 2mLof water,add 3mLof a 10%aqueous solution of potassium hydroxide,and extract this mixture with two 5-mLportions of methylene chloride.Discard the organic phase,heat the aqueous phase on a water bath at 40for 10minutes,cool,acidify with 7%hydrochloric acid,and extract this solution with two 5-mLportions of methylene chloride.Dry the organic phase over anhydrous sodium sulfate,and filter.Evaporate the filtrate to dryness,and dissolve the residue in 1.0mLof methylene chloride.Apply 20µLto the plate.
Standard solution: 0.5mg each of USP Fluorescein RSand USP Valerenic Acid RSper mL,prepared in methanol.Apply 10µLto the plate.
Developing solvent system: a mixture of solvent hexane,ethyl acetate,and glacial acetic acid (65:35:0.5).
Procedure— Spray the plate with anisaldehyde reagent solution,prepared by mixing 0.5mLof anisaldehyde with 10mLof glacial acetic acid,85mLof methanol,and 5mLof sulfuric acid,added in the sequence specified.Heat the plate in an oven at 105for about 10minutes,and examine the plate under white light:the chromatogram of the Standard solutionshows a violet zone due to valerenic acid at an RFvalue of about 0.4,and a yellow zone due to fluorescein at an RFvalue of about 0.1;the chromatogram of the Test solutionshows a violet zone due to valerenic acid at an RFvalue of about 0.4,and a blue-violet zone due to hydroxyvalerenic acid at an RFvalue of about 0.12,just above the yellow zone in the Standard solution;and the chromatogram of the Test solutionmay show other colored zones at RFvalues lower than those of valerenic acid.
B: The retention time of the valerenic acid peak in the chromatogram of the Test preparationcorresponds to that in the chromatogram of the Standard preparation,as obtained in the test for Content of valerenic acid.
Microbial enumeration á2021ñ— It meets 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 1,000cfu per g,the coliform count does not exceed 1,000cfu per g,and the Enterobacteriaceaecount does not exceed 1,000cfu per g.
Loss on drying á731ñ— Dry about 1.0g of the Extract,accurately weighed,at 105for 2hours:it loses not more than 9.0%of its weight.
Total ash á561ñ: not more than 7.0%.
Pesticide residues á561ñ: meets the requirements.
Alcohol content,Method IIá611ñ(if present): not more than 2.0%.
Content of valerenic acid—
Mobile phase— Prepare a mixture of methanol and water (77:27),add 0.5mLof phosphoric acid to each 100mLof the mixture,filter,and degas.Make adjustments if necessary (see System Suitabilityunder Chromatography á621ñ).
Standard preparation— Dissolve an accurately weighed quantity of USP Valerenic Acid RSin methanol,and dilute quantitatively,and stepwise if necessary,with methanol to obtain a solution having a known concentration of about 24µg per mL.
Test preparation— Transfer an accurately weighed quantity of the Extract,equivalent to about 0.6mg of valerenic acid,to a 25-mLvolumetric flask,and add 15mLof methanol.Stir for 10minutes,dilute with methanol to volume,mix,and filter.
Chromatographic system (see Chromatography á621ñ)— The liquid chromatograph is equipped with a 225-nm detector and a 4.6-mm ×25-cm column that contains packing L1.The flow rate is about 1.5mLper minute.The column temperature is maintained at 30.Chromatograph the Standard preparation,and record the peak responses as directed for Procedure:the capacity factor,k¢,determined from valerenic acid is not less than 5;the tailing factor for valerenic acid is not more than 2.0;and the relative standard deviation for replicate injections is not more than 2.0%.
Procedure— Separately inject equal volumes (about 20µL)of the Standard preparationand the Test preparationinto the chromatograph,record the chromatograms,and measure the responses for valerenic acid.Calculate the percentage of valerenic acid (C15H22O2)in the portion of the Extract taken by the formula:
2500(C/W)(rU/rS),
in which Cis the concentration,in mg per mL,of USP Valerenic Acid RSin the Standard preparation;Wis the weight,in mg,of the Extract taken to prepare the Test preparation;and rUand rSare the valerenic acid peak responses obtained from the Test preparationand the Standard preparation,respectively:not less than 0.3%is found.
Auxiliary Information— Staff Liaison:Gabriel I.Giancaspro,Ph.D.,Senior Scientist and Latin American Specialist
Expert Committee:(DSB)Dietary Supplements:Botanicals
USP28–NF23Page 2135
Pharmacopeial Forum:Volume No.27(2)Page 2268
Phone Number:1-301-816-8343