Powdered Valerian Extract
DEFINITION
Powdered Valerian Extract is prepared from comminuted Valerian and with 70% alcohol or other suitable solvents. It contains NLT 0.3% of valerenic acid (C15H22O2). The ratio of the starting crude plant material to the Extract is between 4:1 and 7:1.
IDENTIFICATION
•  A. Thin-Layer Chromatographic Identification Test
Standard solution:  0.5 mg/mL each of USP Fluorescein RS and USP Valerenic Acid RS, in methanol
Sample solution:  Dissolve 0.2 g of Extract in 2 mL of water, add 3 mL of a 10% aqueous solution of potassium hydroxide, and extract this mixture with two 5-mL portions of methylene chloride. Discard the organic phase, heat the aqueous phase on a water bath at 40 for 10 min, cool, acidify with 7% hydrochloric acid, and extract this solution with two 5-mL portions of methylene chloride. Dry the organic phase over anhydrous sodium sulfate, and filter. Evaporate the filtrate to dryness, and dissolve the residue in 1.0 mL of methylene chloride.
Chromatographic system 
Adsorbent:  0.5-mm layer of chromatographic silica gel mixture
Application volume 
Standard solution:  10 µL
Sample solution:  20 µL
Developing solvent system:  Solvent hexane, ethyl acetate, and glacial acetic acid (65: 35: 0.5)
Spray reagent:  Mix 0.5 mL of anisaldehyde with 10 mL of glacial acetic acid, 85 mL of methanol, and 5 mL of sulfuric acid, added in the sequence specified.
Analysis 
Samples:  Standard solution and Sample solution
Spray the plate with Spray reagent. Heat the plate in an oven at 105 for 10 min, and examine the plate under white light.
Acceptance criteria:  The Standard solution chromatogram shows a violet zone due to valerenic acid at an RF value of 0.4, and a yellow zone due to fluorescein at an RF value of 0.1. The Sample solution chromatogram shows a violet zone due to valerenic acid at an RF value of 0.4, and a blue-violet zone due to hydroxyvalerenic acid at an RF value of 0.12, just above the yellow zone in the Standard solution. The chromatogram of the Sample solution may show other colored zones at RF values lower than those of valerenic acid.
•  B. HPLC Identification Test
Analysis:  Proceed as directed in the test for Content of Valerenic Acid.
Acceptance criteria:  The Sample solution chromatogram exhibits a peak at a retention time that corresponds to that of valerenic acid in the Standard solution.
COMPOSITION
•  Content of Valerenic Acid
Mobile phase:  Methanol and water (77:27). Add 0.5 mL of phosphoric acid to each 100 mL of the mixture.
Standard solution:  0.024 mg/mL of USP Valerenic Acid RS in methanol
Sample solution:  Transfer a quantity of the Extract, nominally equivalent to 0.6 mg of valerenic acid, to a 25-mL volumetric flask, and add 15 mL of methanol. Stir for 10 min, dilute with methanol to volume, mix, and filter.
Chromatographic system 
(See Chromatography 621, System Suitability.)
Mode:  LC
Detector:  UV 225 nm
Column:  4.6-mm × 25-cm; packing L1
Column temperature:  30
Flow rate:  1.5 mL/min
Injection size:  20 µL
System suitability 
Sample:  Standard solution
Suitability requirements 
Capacity factor (k¢):  NLT 5 determined from the valerenic acid peak
Tailing factor:  NMT 2.0 for valerenic acid
Relative standard deviation:  NMT 2.0% for the valerenic acid peak
Analysis 
Samples:  Standard solution and Sample solution
Calculate the percentage of valerenic acid (C15H22O2) in the portion of the Extract taken:
Result = (rU/rS) × (CS/CU) × 100
rU== peak area from the Sample solution
rS== peak area from the Standard solution
CS== concentration of USP Valerenic Acid RS in the Standard solution (mg/mL)
CU== concentration of the Extract in the Sample solution (mg/mL)
Acceptance criteria:  NLT 0.3%
CONTAMINANTS
•  Alcohol Determination, Method II 611: NMT 2.0%, if present
•  Microbial Enumeration Tests 2021: The total bacterial count does not exceed 104 cfu/g, the total combined molds and yeasts count does not exceed 103 cfu/g, the coliform count does not exceed 103 cfu/g, and the Enterobacteriaceae count does not exceed 103 cfu/g.
•  Absence of Specified Microorganisms 2022: It meets the requirements of the tests for absence of Salmonella species, Escherichia coli, and Staphylococcus aureus.
SPECIFIC TESTS
•  Loss on Drying 731: Dry 1.0 g of the Extract at 105 for 2 h: it loses NMT 9.0% of its weight.
ADDITIONAL REQUIREMENTS
•  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.
•  USP Reference Standards 11
USP Fluorescein RS Click to View Structure
USP Valerenic Acid RS Click to View Structure
Auxiliary Information— Please check for your question in the FAQs before contacting USP.
Topic/Question Contact Expert Committee
Monograph Maged H. Sharaf, Ph.D.
Principal Scientific Liaison
1-301-816-8318
(DS2010) Monographs - Dietary Supplements
2021 Radhakrishna S Tirumalai, Ph.D.
Principal Scientific Liaison
1-301-816-8339
(GCM2010) General Chapters - Microbiology
2022 Radhakrishna S Tirumalai, Ph.D.
Principal Scientific Liaison
1-301-816-8339
(GCM2010) General Chapters - Microbiology
Reference Standards RS Technical Services
1-301-816-8129
rstech@usp.org
USP35–NF30 Page 1465
Pharmacopeial Forum: Volume No. 27(2) Page 2268