Powdered Rhodiola rosea Extract
DEFINITION
Powdered Rhodiola rosea Extract is prepared from Rhodiola rosea by extraction with hydroalcoholic mixtures. The ratio of plant material to extract is between 1.5: 1 and 5:1. It contains NLT 90.0% and NMT 110.0% of the labeled amount of phenylpropenoid glycosides calculated as the sum of rosarin, rosavin, and rosin, and NLT 90.0% and NMT 110.0% of the labeled amount of salidroside, both calculated on the dried basis. It may contain suitable added substances as carriers.
IDENTIFICATION
• A. Thin-Layer Chromatography
Standard solution A:
1.0 mg/mL of USP Rosavin RS in methanol
Standard solution B:
50 mg/mL of USP Rhodiola rosea Root and Rhizome Dry Extract RS in methanol. Sonicate for 10 min, centrifuge, and use the supernatant.
Sample solution:
50 mg/mL of Powdered Rhodiola rosea Extract in methanol. Sonicate for 10 min, centrifuge, and use the supernatant.
Chromatographic system
Adsorbent:
Chromatographic silica gel with an average particle size of 5 µm (HPTLC plates)
Application volume:
3 µL of Standard solution A and 5 µL each of Standard solution B and the Sample solution; as 8-mm bands
Relative humidity:
Condition the plate to a relative humidity of about 33% using a suitable device.
Developing solvent system:
A mixture of ethyl acetate, methanol, water, and formic acid (77:13:10:2)
Developing distance:
6 cm
Derivatization reagent:
Dissolve 1 g of diphenylamine in 40 mL of acetone, add 1 mL of aniline, and mix. Carefully add 7.5 mL of phosphoric acid, and mix.
Analysis
Samples:
Standard solution A, Standard solution B, and Sample solution
Apply the samples as bands to a suitable high performance thin-layer chromatographic plate, and dry in air. Develop the chromatograms in a saturated chamber, remove the plate from the chamber, and dry in air. Derivatize the plate with Derivatization reagent, heat at 120
for 5 min, and examine under visible light.
for 5 min, and examine under visible light.
System suitability:
The chromatogram of Standard solution B exhibits, in the lower half, three gray bands and two brownish bands, one above and the other below the gray bands; the most intense band in the chromatogram is the brownish band with an RF below the gray bands; the most intense gray band is the lower band at an RF corresponding to the band due to rosavin in the chromatogram of Standard solution A; the upper gray band due to rosarin is less intense.
Acceptance criteria:
The chromatogram of the Sample solution exhibits a gray band corresponding to the band due to rosavin in the chromatogram of Standard solution A, and the following bands corresponding to similar bands in the chromatogram of Standard solution B: two additional gray bands and two brownish bands, one above and the other below the gray bands; the most intense band in the chromatogram is the brownish band with an RF below the gray bands; the most intense gray band is the lower band due to rosavin.
• B. HPLC
Analysis:
Proceed as directed in the test for Content of Phenylpropenoid Glycosides and Salidroside.
Acceptance criteria:
The chromatogram of the Sample solution exhibits peaks at the retention times corresponding to the peaks due to salidroside, tyrosol, rosarin, rosavin, rosin, and rosiridin in the chromatogram of Standard solution C. The ratio of the contents of rosarin, rosavin, and rosin is about 2.5: 6.0: 1.5, respectively.
COMPOSITION
• Content of Phenylpropenoid Glycosides and Salidroside
Solution A:
Water
Solution B:
Acetonitrile
Mobile phase:
See Table 1.
Table 1
| Time (min) |
Solution A (%) |
Solution B (%) |
|---|---|---|
| 0 | 94 | 6 |
| 6 | 83 | 17 |
| 7 | 80.3 | 19.7 |
| 9 | 80.3 | 19.7 |
| 10 | 0 | 100 |
| 12 | 94 | 6 |
| 17 | 94 | 6 |
Standard solution A:
1.0 mg/mL of USP Rosavin RS in methanol
Standard solution B:
0.3 mg/mL of USP Salidroside RS in methanol
Standard solution C:
4.0 mg/mL of USP Rhodiola rosea Root and Rhizome Dry Extract RS in methanol. Sonicate to dissolve, if necessary. Before injection, pass through a membrane filter of 0.45-µm or finer pore size.
Sample solution:
4.0 mg/mL of Powdered Rhodiola rosea Extract in methanol. Sonicate to dissolve, if necessary. Before injection, pass through a membrane filter of 0.45-µm or finer pore size.
Chromatographic system
Mode:
LC
Detector:
UV 205 nm
Column:
3.0-mm × 10-cm; 2.5-µm packing L1
Column temperature:
40 ± 1
Flow rate:
1.0 mL/min
Injection volume:
1 µL
System suitability
Samples:
Standard solution A and Standard solution C
Suitability requirements
Chromatogram similarity:
The chromatogram obtained from Standard solution C is similar to the reference chromatogram provided with the lot of USP Rhodiola rosea Root and Rhizome Dry Extract RS being used.
Resolution:
NLT 1.5 between the rosarin and rosavin peaks, Standard solution C
Relative standard deviation:
NMT 2% determined from the rosavin peak in repeated injections, Standard solution A
Analysis
Samples:
Standard solution A, Standard solution B, Standard solution C, and Sample solution
Using the chromatograms of Standard solution A, Standard solution B, Standard solution C, and the reference chromatogram provided with the lot of USP Rhodiola rosea Root and Rhizome Dry Extract RS being used, identify the retention time of the peaks corresponding to salidroside, tyrosol, rosarin, rosavin, rosin, and rosiridin from the Sample solution.
Separately calculate the percentage of rosarin, rosavin, and rosin as rosavin in the portion of Powdered Rhodiola rosea Extract taken:
P1 = (rU/rS) × (CS/CU) × 100
| rU | = | = peak area of the relevant analyte from the Sample solution |
| rS | = | = peak area of rosavin from Standard solution A |
| CS | = | = concentration of rosavin in Standard solution A (mg/mL) |
| CU | = | = concentration of Powdered Rhodiola rosea Extract in the Sample solution (mg/mL) |
Calculate the percentage of phenylpropenoid glycosides as the sum of the percentages of rosarin, rosavin, and rosin.
Calculate the percentage of salidroside in the portion of Powdered Rhodiola rosea Extract taken:
P2 = (rU/rS) × (CS/CU) × 100
| rU | = | = peak area of salidroside from the Sample solution |
| rS | = | = peak area of salidroside from Standard solution B |
| CS | = | = concentration of salidroside in Standard solution B (mg/mL) |
| CU | = | = concentration of Powdered Rhodiola rosea Extract in the Sample solution (mg/mL) |
Calculate the percentage of the labeled amount of phenylpropenoid glycosides in the portion of Powdered Rhodiola rosea Extract taken:
Result = (P1/L) × 100
| P1 | = | = content of phenylpropenoid glycosides, as determined above (%) |
| L | = | = labeled amount of phenylpropenoid glycosides (%) |
Calculate the percentage of the labeled amount of salidroside in the portion of Powdered Rhodiola rosea Extract taken:
Result = (P2/L) × 100
| P2 | = | = content of salidroside, as determined above (%) |
| L | = | = labeled amount of salidroside (%) |
Acceptance criteria:
NLT 90.0% and NMT 110.0% of the labeled amount of phenylpropenoid glycosides, and NLT 90.0% and NMT 110.0% of the labeled amount of salidroside, both calculated on the dried basis.
CONTAMINANTS
• Elemental Impurities—Procedures 
233
233
Acceptance criteria
Arsenic:
NMT 2.0 µg/g
Cadmium:
NMT 1.0 µg/g
Lead:
NMT 5.0 µg/g
Mercury:
NMT 1.0 µg/g
• Articles of Botanical Origin, General Method for Pesticide Residues Analysis 561:
Meets the requirements
561:
Meets the requirements
• Microbial Enumeration Tests 2021:
The total aerobic bacterial count does not exceed 104 cfu/g, and the total combined molds and yeasts count does not exceed 103 cfu/g.
2021:
The total aerobic bacterial count does not exceed 104 cfu/g, and the total combined molds and yeasts count does not exceed 103 cfu/g.
• Absence of Specified Microorganisms 2022:
Meets the requirements of the tests for absence of Salmonella species and Escherichia coli
2022:
Meets the requirements of the tests for absence of Salmonella species and Escherichia coli
• Articles of Botanical Origin, Test for Aflatoxins 561:
Meets the requirements
561:
Meets the requirements
SPECIFIC TESTS
• Loss on Drying 731
731
Sample:
2.0 g of Powdered Rhodiola rosea Extract
Analysis:
Dry at 105 for 2 h.
for 2 h.
Acceptance criteria:
NMT 5%
ADDITIONAL REQUIREMENTS
• Packaging and Storage:
Preserve in well-closed containers, protected from light and moisture, and store at controlled room temperature.
• Labeling:
The label states the Latin binomial and, following the official name, the part of the plant from which the article was derived. It meets other labeling requirements under Botanical Extracts 565.
565.
• USP Reference Standards 11
11
USP Rhodiola rosea Root and Rhizome Dry Extract RS
USP Rosavin RS 
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USP Salidroside RS
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Auxiliary Information—
Please check for your question in the FAQs before contacting USP.
| Topic/Question | Contact | Expert Committee |
|---|---|---|
| Monograph | Anton Bzhelyansky, M.S.
Scientific Liaison (301) 230-6303 |
(DS2010) Monographs - Dietary Supplements and Herbal Medicines |
| 2021 |
Radhakrishna S Tirumalai, Ph.D.
Principal Scientific Liaison (301) 816-8339 |
(GCM2010) General Chapters - Microbiology |
| 2022 |
Radhakrishna S Tirumalai, Ph.D.
Principal Scientific Liaison (301) 816-8339 |
(GCM2010) General Chapters - Microbiology |
| Reference Standards | RS Technical Services 1-301-816-8129 rstech@usp.org |
USP38–NF33 Page 6198
Pharmacopeial Forum: Volume No. 39(3)