Powdered Valerian - US Pharmacopeial Convention

. ivatization
Revision Bulletin
Official December 1, 2013 Valerian 1
reagent B, heat at 100° for 3 min, and
Powdered Valerian
examine under white light.
Acceptance criteria: After treatment with Derivatization
DEFINITION
reagent A and heating, the Sample solution does
not exhibit an intense blue band at about the middle
Change to read:
of the chromatogram nor any other significant bands
[distinction from Mexican valerian (Valeriana edulis)],
though minor bands may be observed.
Powdered Valerian is Valerian reduced to a fine or a very
After treatment with Derivatization reagent B and heatfine
powder. It contains no calcium oxalate crystals and
ing, the Sample solution exhibits three violet bands in
no foreign starch granules. It contains NLT 0.3% of vola- positions and colors similar to the bands of Standard
tile oil, NLT 0.04% of valerenic acid (C 15H 22O 2), ■ .and NLT solution B. These bands include a minor band in the
0.1% of total valerenic acids, calculated as the sum of lower third of the chromatogram due to hydroxhydroxyvalerenic
acid, acetoxyvalerenic acid, and valer-
yvalerenic acid, a major band at about the middle the
enic acid, on the dried basis. ■1S (USP36)
chromatogram due to acetoxyvalerenic acid [distinction
IDENTIFICATION
from Scouler’s valerian (Valeriana wallichii)], and a
major band at an R F corresponding to the valerenic
Change to read:
acid band of Standard solution A and Standard solution
B. Other minor bands may be observed in the Sample
• A. ■.Meets the requirements for Specific Tests, Botanic
solution and in Standard solution B. ■1S (USP36)
Characteristics ■1S (USP36)
Add the following:
Change to read:
■
.• D. HPLC
Analysis: Proceed as directed in the test for Content of
Valerenic Acids.
Acceptance criteria: The Sample solution exhibits a
peak at a retention time corresponding to the valerenic
acid peak of Standard solution A. The Sample solution
shows additional peaks corresponding to hydroxyvaler-
enic acid and acetoxyvalerenic acid. ■1S (USP36)
• B.
■
.Sample solution: 0.2 g of Powdered Valerian in 5 mL
of methylene chloride. Shake several times, and allow
to stand for 5 min. Filter, wash the filter with 2 mL of
methylene chloride, combine the filtrate and washings,
and evaporate to dryness. Dissolve the residue in
0.2 mL of methylene chloride.
Analysis: To 0.1 mL of the Sample solution add 3 mL of COMPOSITION
a mixture of equal volumes of glacial acetic acid and
25% hydrochloric acid, and shake several times. Change to read:
Acceptance criteria: A blue color develops within 15
min. ■1S (USP36)
Add the following:
■
.• C. THIN-LAYER CHROMATOGRAPHY
• CONTENT OF VALERENIC ■ .ACIDS ■1S (USP36)
■
.Solution A: Mix 6 mL of 85% phosphoric acid with
900 mL of water, dilute with water to 1000 mL, mix,
filter, and degas.
Solution B: Mix 6 mL of 85% phosphoric acid with
900 mL of methanol, dilute with methanol to 1000 mL,
mix, filter, and degas.
Standard solution A: 0.25 mg/mL of USP Valerenic
Acid RS in methanol
Standard solution B: 40 mg/mL of USP Powdered Va- Mobile phase: See Table 1.
lerian Extract RS in methanol. Sonicate for 10 min,
centrifuge, and use the supernatant.
Sample solution: About 0.5 g of Powdered Valerian in
Table 1
5 mL of methanol. Sonicate for 10 min, centrifuge, and Time Solution A Solution B
use the supernatant. (min) (%) (%)
Chromatographic system 0 40 60
Adsorbent: Chromatographic silica gel mixture with
an average particle size of 2–10 µm (HPTLC plates)
Application volume: 5 µL, as 8-mm bands
Developing solvent system: A mixture of cyclohexane,
ethyl acetate, and acetic acid (60:38:2)
Derivatization reagent A: A mixture of glacial acetic
acid and hydrochloric acid (1:4)
Derivatization reagent B: 0.5 mL of p-anisaldehyde,
10 mL of acetic acid, and 5 mL of sulfuric acid. Add to
85 mL of ice-cold methanol, and mix.
Analysis
Samples: Standard solution A, Standard solution B, and
Sample solution
Apply the Samples as bands to a suitable high-perfor-
mance thin-layer chromatographic plate. Use a saturated
chamber, and condition the plate to a relative
humidity of about 33% using a suitable device. Develop
the chromatograms over a distance of 6 cm.
Remove the plate from the chamber, dry, derivatize
with Derivatization reagent A, heat at 120° for 5 min,
and examine under white light. Derivatize with Der-
15 5 95
25 5 95
30 40 60
Solvent: A mixture of methanol and a solution of 0.1%
phosphoric acid in water (3:1)
Standard solution A: 0.02 mg/mL of USP Valerenic
Acid RS in methanol. Sonicate if necessary.
Standard solution B: Sonicate a portion of USP Powdered
Valerian Extract RS in Solvent to obtain a solution
having a concentration of about 20 mg/mL. Before injection,
pass through a membrane filter of 0.45-µm or
finer pore size, discarding the first few mL of the
filtrate.
Sample solution: To a 50-mL volumetric flask, transfer
about 1.0 g of Powdered Valerian, accurately weighed,
add 10.0 mL of water, and shake for 2 min while heat-
ing in a water bath maintaned at about 50°. Sonicate
for 15 min, add 35 mL of methanol, and sonicate for
15 min. Cool, dilute with methanol to volume, and
mix. Before injection, pass through a membrane filter
©2013 The United States Pharmacopeial Convention All Rights Reserved.

Revision Bulletin
2 Valerian Official December 1, 2013
of 0.45-µm or finer pore size, discarding the first few Sample: 100 g of Powdered Valerian
mL of the filtrate. Acceptance criteria: NLT 0.3%
Chromatographic system
(See Chromatography 〈621〉, System Suitability.)
CONTAMINANTS
Mode: LC
Detector: UV 225 nm
Delete the following:
Column: 4.6-mm × 25-cm; end-capped, 5-µm 100 Å
packing L1
•.• HEAVY METALS 〈231〉: 50 µg/g
Column temperature: 40°
•.• (RB 1-Dec-2013) • (RB 1-Dec-2013)
Flow rate: 1.0 mL/min
Injection volume: 25 µL
Change to read:
System suitability
Samples: Standard solution A and Standard solution B •.• ELEMENTAL IMPURITIES—PROCEDURES 〈233〉
Suitability requirements
Acceptance criteria
Chromatogram similarity: The chromatogram of
Arsenic: NMT 0.5 µg/g
Standard solution B is similar to the reference chro-
Cadmium: NMT 1.0 µg/g
matogram provided with the lot of USP Powdered
Lead: NMT 5.0 µg/g
Valerian Extract RS being used.
Mercury: NMT 0.1 µg/g• (RB 1-Dec-2013)
Tailing factor: NMT 2.0 for the valerenic acid peak, •.• (RB 1-Dec-2013)
Standard solution A
• ARTICLES OF BOTANICAL ORIGIN, General Method for Pesti-
Relative standard deviation: NMT 2.0% for the
cide Residues Analysis 〈561〉: Meets the requirements
valerenic acid peak in repeated injections, Standard
• MICROBIAL ENUMERATION TESTS 〈2021〉: The total bactesolution
A
rial count does not exceed 10 cfu/g, the total com-
Analysis
5. bined molds and yeasts count does not exceed 10 cfu/
Samples: Standard solution A, Standard solution B, and
3. g, and bile-tolerant Gram-negative bacteria does not ex-
Sample solution
ceed 10 cfu/g.
Identify the valerenic acids in the Sample solution chro-
3. • ABSENCE OF SPECIFIED MICROORGANISMS 〈2022〉: It meets
matogram by comparison with the chromatograms of
the requirements of the tests for absence of Salmonella
Standard solution A, Standard solution B, and the referspecies
and Escherichia coli.
ence chromatogram provided with the lot of USP
Powdered Valerian Extract RS being used.
SPECIFIC TESTS
Calculate the percentages of hydroxyvalerenic acid,
acetoxyvalerenic acid, and valerenic acid in the por- Change to read:
tion of Powdered Valerian taken:
• BOTANIC CHARACTERISTICS
Result = (r U/r S) × C S × (V/W) × F × 100
Microscopic: ■ .Numerous starch granules, simple, hilum
dotted, stellate or cleft-shaped, 5–15 µm in diamer
U = peak area of the relevant analyte from the
ter; compound starch granules of 2–6 components;
Sample solution
black, cruciate shape under a polarizing microscope.
r S = peak area of valerenic acid from Standard
Scattered stone cells, single or aggregated, with cell
solution A
lumina of various sizes, bright yellowish-white or bright
C S = concentration of valerenic acid in Standard
white under a polarizing microscope; numerous fragsolution
A (mg/mL)
ments of parenchyma cells containing globules of vola-
V = volume of the Sample solution (mL)
tile oil and starch granules; fragments of pale yellow
W = weight of Powdered Valerian taken to prepare
lignified fibers, scattered, single or aggregated; fragthe
Sample solution (mg)
ments of vessels, reticulate, bordered pitted and spiral.
F = conversion factor for each analyte (1.10 for
■1S (USP36)
hydroxyvalerenic acid, 1.25 for
acetoxyvalerenic acid, and 1.00 for valerenic
Delete the following:
acid)
Acceptance criteria: NLT 0.04% of valerenic acid
■
.• ARTICLES OF BOTANICAL ORIGIN, Foreign Organic Matter
(C 15H 22O 2), and NLT 0.1% of total valerenic acids, cal-
〈561〉: NMT 2.0% culated as the sum of hydroxyvalerenic acid, acetox-
■1S (USP36)
• EXTRACTABLE MATTER
yvalerenic acid, and valerenic acid on the dried
Sample: 2 g of Powdered Valerian, carefully dried at
basis ■1S (USP36) 40°
• ARTICLES OF BOTANICAL ORIGIN, Volatile Oil Determination
Analysis: Mix the Sample with 20 mL of 70% alcohol,
〈561〉
and allow to stand for 2 h, shaking frequently. Filter,
evaporate 5 mL of the filtrate on a water bath to dryness,
and dry the residue at 105°.
Acceptance criteria: NLT 20%
Delete the following:
■
.• WATER, Method 1a 〈921〉: NMT 5.0% ■1S (USP36)
©2013 The United States Pharmacopeial Convention All Rights Reserved.

Revision Bulletin
Official December 1, 2013 Valerian 3
Add the following:
• LABELING: The label states the Latin binomial and, following
the official name, the parts of the plant from
■
.• LOSS ON DRYING 〈731〉
which the article was derived.
Sample: 1.0 g of Powdered Valerian
Analysis: Dry at 105° for 2 h.
Change to read:
Acceptance criteria: NMT 12% ■1S (USP36)
• ARTICLES OF BOTANICAL ORIGIN, Total Ash 〈561〉: NMT • USP REFERENCE STANDARDS 〈11〉
12.0% ■.USP Powdered Valerian Extract RS ■1S (USP36)
• ARTICLES OF BOTANICAL ORIGIN, Acid-Insoluble Ash 〈561〉: USP Valerenic Acid RS
NMT 5.0%
ADDITIONAL REQUIREMENTS
• PACKAGING AND STORAGE: Preserve in well-closed containers,
store at room temperature, and protect from
light and moisture.
©2013 The United States Pharmacopeial Convention All Rights Reserved.